Showing papers in "Medical Oncology in 2014"

The prognostic value of preoperative NLR, d-NLR, PLR and LMR for predicting clinical outcome in surgical colorectal cancer patients

Abstract: Accumulating evidences indicate cancer-triggered inflammation plays a pivotal role in carcinogenesis. Systematic inflammatory response biomarkers are considered as potential prognostic factors for improving predictive accuracy in colorectal cancer (CRC). Preoperative neutrophil-to-lymphocyte ratio (NLR), derived neutrophil-to-lymphocyte ratio (d-NLR), platelet-to-lymphocyte ratio (PLR) and lymphocyte- to-monocyte ratio (LMR) were investigated and compared in 205 surgical CRC patients. ROC curve was applied to determine thresholds for four biomarkers, and their prognostic values were assessed using Kaplan–Meier curve, univariate and multivariate COX regression models. Moreover, a number of risk factors were used to form nomograms for evaluating risk of survival, and Harrell’s concordance index (c-index) was used to evaluate predictive accuracy. Results showed that elevated NLR was significantly associated with diminished recurrent-free survival (RFS), overall survival (OS) and cancer-specific survival (CSS) in surgical CRC patients. Moreover, multivariate COX analysis identified elevated NLR as an independent factor for poor RFS (P < 0.001, HR 2.52, 95 % CI 1.65–3.83), OS (P < 0.001, HR 2.73, 95 % CI 1.74–4.29) and CSS (P < 0.001, HR 2.77, 95 % CI 1.72–4.46). Additionally, predictive nomograms including NLR for RFS, OS and CSS could be more effective in predicting RFS (c-index: 0.810 vs. 0.656), OS (c-index: 0.809 vs. 0.690) and CSS (c-index: 0.802 vs. 0.688) in surgical CRC patients, respectively. These findings indicate that preoperative elevated NLR can be considered as an independent prognostic biomarker for RFS, OS and CSS. Nomograms containing NLR provide improved accuracy for predicting clinical outcomes in surgical CRC patients under surgery resection.

Combined detection of serum exosomal miR-21 and HOTAIR as diagnostic and prognostic biomarkers for laryngeal squamous cell carcinoma

Abstract: Serum exosomes containing noncoding RNA (ncRNA) play an important role in both physiological and pathological conditions. However, biological function of exosomal ncRNA remains unclear. The aim of the study was to investigate the prognostic and diagnostic values of exosomal ncRNA by comparing the amounts of exosomal miR-21 and HOTAIR in serum of laryngeal squamous cell carcinoma (LSCC) patients with those of polyps of vocal cords, and by determinating whether combined detection of the two molecules could provide useful information in the diagnosis of LSCC. Exosomes were isolated from the serum samples of 52 LSCC patients and those of 49 patients with polyps of vocal cords. TEM and Western blot were applied for the confirmation of isolated exosomes by observing the ultra structure and testing CD63 marker protein, respectively. RT-PCR was performed to detect the expression of miR-21 and HOTAIR in the exosomes. The receiver-operating characteristic (ROC) curve was generated to examine the prognostic value of the two molecules. The expression of exosomal miR-21 and HOTAIR was significantly higher in patients with LSCC than those with vocal cord polyps. There were significant differences of serum exosomal miR-21 and HOTAIR expressions between the advanced T classifications (T3/T4) or clinical stages (III/IV) and the early stages. The patients with lymph node metastasis had higher serum exosomal miR-21 and HOTAIR expressions than those without. There were no differences between patient sex, tumor locations and differentiations. The area under the ROC curve of combined examination of exosomal HOTAIR and miR-21 for diagnosing LSCC was 87.6 %, which was significantly higher than 80.1 % of miR-21 (p = 0.0359) or 72.7 % of HOTAIR (p = 0.0012), showing 94.2 and 73.5 % of sensitivity and specificity, respectively, in differentiating the malignant from benign laryngeal disease. Serum exosomal miR-21 and HOTAIR were significantly correlated with clinical parameters of LSCC, and combined evaluation of their serum expressions may be a valuable biomarker to screen LSCC and might be a promising predicting tool for LSCC patient.

Insulin receptor (IR) and insulin-like growth factor receptor 1 (IGF-1R) signaling systems: novel treatment strategies for cancer

Abstract: Insulin and insulin-like growth factor (IGF) signaling system, commonly known for fine-tuning numerous biological processes, has lately made its mark as a much sought-after therapeutic targets for diabetes and cancer These receptors make an attractive anticancer target owing to their overexpression in variety of cancer especially in prostate and breast cancer Inhibitors of IGF signaling were subjected to clinical cancer trials with the main objective to confirm the effectiveness of these receptors as a therapeutic target However, the results that these trials produced proved to be disappointing as the role played by the cross talk between IGF and insulin receptor (IR) signaling pathways at the receptor level or at downstream signaling level became more lucid Therapeutic strategy for IGF-1R and IR inhibition mainly encompasses three main approaches namely receptor blockade with monoclonal antibodies, tyrosine kinase inhibition (ATP antagonist and non-ATP antagonist), and ligand neutralization via monoclonal antibodies targeted to ligand or recombinant IGF-binding proteins Other drug-discovery approaches are employed to target IGF-1R, and IR includes antisense oligonucleotides and recombinant IGF-binding proteins However, therapies with monoclonal antibodies and tyrosine kinase inhibition targeting the IGF-1R are not evidenced to be satisfactory as expected Factors that are duly held responsible for the unsuccessfulness of these therapies include (a) the existence of the IR isoform A overexpressed on a variety of cancers, enhancing the mitogenic signals to the nucleus leading to the endorsement of cell growth, (b) IGF-1R and IR that form hybrid receptors sensitive to the stimulation of all three IGF axis ligands, and (c) IGF-1R and IR that also have the potential to form hybrid receptors with other tyrosine kinase to potentiate the cellular transformation, tumorigenesis, and tumor vascularization This mini review is a concerted effort to explore and fathom the well-recognized roles of the IRA signaling system in human cancer phenotype and the main strategies that have been so far evaluated to target the IR and IGF-1R

Long noncoding RNA GAS5 affects cell proliferation and predicts a poor prognosis in patients with colorectal cancer

Abstract: Colorectal cancer (CRC) is the third most common type of cancer worldwide. Recent studies have shown that lncRNAs play important roles in carcinogenesis. The aim of this study was to explore the role of lncRNA GAS5 in CRC. Real-time PCR was performed to investigate the expression of GAS5 in tumor tissues and corresponding non-tumor colorectal tissues from 66 patients with CRC. The lower expression of GAS5 was significantly correlated with large tumor size, low histological grade and advanced TNM stage. Multivariate analyses revealed that GAS5 expression served as an independent predictor for overall survival (P = 0.034). Further experiments revealed that overexpressed GAS5 significantly repressed the proliferation both in vitro and in vivo. In conclusion, our results suggest that GAS5, as a growth regulator, may serve as a candidate prognostic biomarker in human colorectal cancer.

Retraction Note to: A randomized phase II study of everolimus for advanced pancreatic neuroendocrine tumors in Chinese patients

Abstract: Everolimus, an oral inhibitor of mammalian target of mTOR, has been recently shown to have antitumor effect in a phase III, double-blind, randomized trial (RADIANT-3) of 410 patients with advanced pancreatic neuroendocrine tumors (PNETs). The purpose of this study was to investigate the specific efficacy and safety of everolimus in the Chinese patient with PNETs. In this randomized phase II study, the analysis on Chinese patients was performed comparing efficacy and safety between everolimus 10 mg/day orally (n = 44) and matching placebo (n = 35). The primary endpoint was progression-free survival (PFS). Adverse events were also examined. The median PFS was 15.47 months with everolimus [95 % confidence interval (CI) 10.52–26.37], as compared to 4.29 months with placebo (95 % CI 2.22–10.75), representing a 72 % reduction in the risk of progression or death (hazard ratio 0.27; 95 % CI 0.13–0.59; P < 0.001). Drug-associated adverse events (AEs) were mostly grade 1 or 2, observed in all 44 (100 %) patients receiving everolimus and in 29 (83 %) patients receiving placebo. The most common AEs (grade 1–4) associated with everolimus were rash (n = 38; 86 %), stomatitis (n = 30; 68 %), infections (n = 33; 75 %), epistaxis (n = 32; 73 %), pneumonitis (n = 27; 61 %) and anemia (n = 22; 50 %). Everolimus when compared with placebo is effectively in improving PFS in Chinese patients with PNETs.

Long noncoding RNA HULC is a novel biomarker of poor prognosis in patients with pancreatic cancer

Abstract: Pancreatic cancer (PC) is the fourth leading cause of cancer-related mortalities in the USA and the sixth leading cause of mortality in China. Recent studies have shown that lncRNAs play important roles in carcinogenesis. The aim of this study was to explore the role of lncRNA HULC in PC. Quantitative real-time PCR was performed to investigate the expression of HULC in tumor tissues and corresponding normal tissues from 304 patients with PC. The higher expression of HULC was significantly correlated with large tumor size, advanced lymph node metastasis and vascular invasion. Multivariate analyses revealed that HULC expression served as an independent predictor for overall survival (P = 0.032). Further experiments revealed that HULC knockdown significantly repressed cell proliferation of PC in vitro. In conclusion, our results suggest that HULC may serve as a candidate prognostic biomarker through growth regulation in human PC.

c-Myc-induced, long, noncoding H19 affects cell proliferation and predicts a poor prognosis in patients with gastric cancer.

Abstract: Gastric cancer (GC) is one of the most frequent cancers worldwide. Recent studies have shown that long noncoding RNAs (lncRNAs) play critical roles in multiple biological processes, including oncogenesis. The present study aimed to evaluate the potential role of lncRNA H19 in GC. qRT-PCR was performed to investigate the expression of H19 in tumor tissues and corresponding non-tumor lung tissues from 80 patients with GC and in GC cell lines. The Kaplan-Meier method and Cox proportional hazards analysis were used to evaluate the association between H19 expression and overall survival time (OS). The biological significance of H19 was evaluated using siRNAs in vitro. We also constructed a c-Myc plasmid to investigate the cause of the altered expression of H19 in the progression of GC. The results show that lncRNA H19 is overexpressed in tumor tissues compared with adjacent normal tissues. An advanced tumor-node-metastasis stage was positively correlated with increased H19 expression (P < 0.001), and a high H19 expression was associated with poor OS and can be regarded as an independent predictor of the OS of GC patients (P = 0.042). MTT and colony formation assays confirmed that H19 expression affects GC cell proliferation in vitro. Furthermore, exogenous c-Myc significantly induces H19 expression, and the expression of H19 was positively correlated with the c-Myc levels in the 80 samples used in our study (Pearson correlation coefficient = -0.687). In conclusion, our study demonstrates that the altered expression of lncRNA H19, which is induced by c-Myc, is involved in the development and progression of GC by regulating cell proliferation and shows that H19 may be a potential diagnostic and prognostic target in patients with GC.

Granulocyte–macrophage colony-stimulating factor: not just another haematopoietic growth factor

Abstract: Granulocyte-macrophage colony-stimulating factor (GM-CSF) is often used to treat leucopenia. Other haematopoietins may increase the number of circulating leucocytes with higher efficiency, but GM-CSF has additional effects that may be far more relevant than its haematopoietic activity. GM-CSF induces differentiation, proliferation and activation of macrophages and dendritic cells which are necessary for the subsequent T helper cell type 1 and cytotoxic T lymphocyte activation. GM-CSF haematopoietic and non-haematopoietic functions have pro-inflammatory and immune regulatory potential to treat a variety of autoimmune diseases and tumours. On the other hand, GM-CSF deficiency leads to various immune dysfunctions and the current utilization of GM-CSF as haematopoietic factor might be an accurate but very incomplete indication for a cytokine with vast clinical potential.

MiR-148a regulates MEG3 in gastric cancer by targeting DNA methyltransferase 1.

Abstract: The long non-coding RNA MEG3 has been reported to be a tumor suppressor in a number of malignant tumors including gastric cancer. Several studies have shown that the regulation of MEG3 may attribute to the promoter hypermethylation. However, the mechanism of MEG3 regulation in gastric cancer is still not well understood. MiR-148a can suppress gastric tumorigenesis through regulating the expression of target genes such as DNA methyltransferase 1(DNMT-1). We examined the expression of MEG3 in 52 gastric cancer samples using quantitative real-time PCR and found the down-regulation of MEG3 in both gastric cancer tissues and cell lines. The positive correlation of MEG3 and miR-148a was further confirmed in SGC-7901 and BGC-823 gastric cancer cell lines. Hypermethylation of MEG3 differentially methylated regions was identified by methylation-specific PCR, and MEG3 expression was increased with the inhibition of methylation with siRNA to DNMT-1 in gastric cancer cells. In addition, transfection of MEG3 siRNA into gastric cancer cells diminished the suppression of proliferation induced by overexpression of miR-148a. Our results suggest that the suppression of miR-148a may contribute to the down-regulation of MEG3 in gastric cancer by modulation of DNMT-1.

Abnormal expression of EMT-related proteins, S100A4, vimentin and E-cadherin, is correlated with clinicopathological features and prognosis in HCC

Abstract: We determined the expression of epithelial-mesenchymal transition (EMT) indicator proteins, E-cadherin (E-cad), vimentin (VIM), mucin 1 (MUC1) and S100 calcium-binding protein A4 (S100A4) in hepatocellular carcinoma (HCC) patient tissue samples. We also investigated the relationship between the expression of these proteins and clinicopathologic factors in HCC. Finally, we assessed the potential value of these markers as prognostic indicators of survival in HCC patients. The expression of E-cad, VIM, MUC1 and S100A4 EMT indicator proteins was assessed in tissue microarray HCC tissue sections and corresponding peritumoral normal tissues by immunohistochemistry. In addition, the expression for the four EMT indicator proteins was correlated with clinicopathological features of HCC and patient outcome. Comparison of clinicopathological characteristics and immunohistochemistry by χ2 analysis revealed that downregulation of E-cad in HCC was significantly associated with later TNM cancer stage (P = 0.012), gross classification (P = 0.018), regional lymph node metastasis (P = 0.036) and liver cirrhosis (P = 0.028). Increased S100A4 expression in HCC was significantly associated with differentiation (P = 0.032), tumor with a complete fibrous capsule (P = 0.031) and portal vein invasion (P = 0.038). High VIM expression in HCC was significantly associated with high serum α-fetoprotein levels (P = 0.016). We also observed that low E-cad expression was significantly associated with overexpression of VIM (P = 0.001). Kaplan–Meier survival and Cox regression analysis revealed that low E-cad expression (HR = 0.164, 95 % CI 0.072 to 0.373, P < 0.001) and high serum α-fetoprotein levels (HR = 2.202, 95 % CI 1.054 to 4.598, P = 0.036) were independent prognostic factors in HCC. Our study demonstrates that high S100A4 and VIM expression and low E-cad expression correlate with an aggressive, malignant phenotype in HCC. These results also support a role for E-cad as a prognostic factor in HCC.

The effect of immune microenvironment on the progression and prognosis of colorectal cancer

Abstract: Cancer cells may escape from host immune responses through active suppression of the immune response, but the detailed mechanisms in colorectal cancer remain to be elucidated. In this study, 108 colorectal tumor samples and their peritumoral tissues were collected for immunohistochemistry of infiltrating lymphocytes. Th1 and Tregs cells were determined by the positive expression of T-bet and FOXP3 proteins, respectively. The Tr1 cells were identified by CD49b and LAG-3 protein expression. IL-17-positive cells were identified by IL-17 expression. Results showed that the percentage of T-bet-positive cells was significantly decreased, while the percentages of IL-17-, FOXP3-, CD49b-, and LAG-3-positive cells were significantly increased in tumor tissues compared to that in peritumoral tissues. The ratio of IL-17-, FOXP3-, CD49b-, and LAG3-positive cells to T-bet-positive cells was significantly higher in tumor tissues than in peritumoral tissues. The percentage of infiltrating IL-17-positive cells in tumor tissues was negatively associated with lymph metastasis, invasion, and TNM stage. The percentage of CD49b- and LAG-3-positive cells was positively associated with differentiation, lymph metastasis, invasion, TNM, and Duke stage of colorectal cancer. The percentage of positive Th17 and Tr1 cells is a marker for poor prognosis in patients with CRC. In conclusion, decreased composition of regulative Th1 cells and increased composition of FOXP+ Tregs-, CD49b+/LAG-3+ Tr1-, and IL-17-positive cells in tumor tissues may be associated with the progression of colorectal cancer. The high percentage of IL-17- and Tr1-positive cells in tumor tissues is a predictive marker for poor prognosis of colorectal cancer.

Benign reversible encephalopathy syndrome after bevacizumab therapy for metastatic ovarian cancer

Abstract: Posterior reversible encephalopathy syndrome (PRES) is now a well-described entity in neurology, diagnosed by the clinical picture of altered mental status, headache, visual disturbances, hypertension and seizures, associated with the classical MRI findings of cortical and subcortical abnormalities in multiple brain areas, predominantly in the posterior lobes. This entity has been associated with eclampsia, hypertensive encephalopathy, neurotoxic drugs, immunosuppressants and electrolyte disturbances [1]. We describe a case of PRES, following bevacizumab therapy in a patient with metastatic ovarian carcinoma. We are presenting this case to highlight the benign nature of the seizure component of this syndrome and to suggest modifying its nomenclature to benign reversible encephalopathy syndrome (BRES) because most of the reported cases in the literature describe lesion in the frontal and temporal lobes and not only the posterior lobes and the prognosis for recovery from this condition is very good. A 31-year-old female with metastatic high-grade endometrioid ovarian squamous cell carcinoma on multiple chemotherapy regimens presented with a focal tonic–clonic seizure activity with secondary generalization 16 h after receiving intravenous bevacizumab for the first time. The seizures were controlled by intravenous benzodiazepines, phenytoin and valproic acid. The patient was not known to have any previous neurological disorder or metastatic cerebral disease. Neurological examination was consistent with a postictal state, following benzodiazepine treatment. Cerebrospinal fluid analysis was normal with no malignant cells. MRI of the brain showed abnormal high signal intensity on T2 and FLAIR sequences in the cortical and subcortical matter of the parietal and occipital lobes, posterior fossa, as well as the left pons, left cerebral peduncle and both frontal lobes (Fig. 1). EEG revealed severe generalized cortical slowing with no ongoing epileptiform activity. The patient recovered slowly over a fortnight and was discharged home seizure free to continue treatment for her ovarian cancer. PRES is an acute encephalopathy with diverse neurologic symptoms including headache, visual disturbance, altered mental status, hypertension and seizures [1]. It is diagnosed by the clinical setting associated with typical MRI changes in hyperintense lesion on T2 and FLAIR sequences in multiple regions of the brain, predominantly in the posterior lobes and posterior fossa [1, 2]. It has been associated with eclampsia, hypertensive crisis, chemotherapeutic agents, and immunosuppressant therapy and electrolyte disturbances [1, 2]. The pathophysiology of this syndrome is not clearly defined. One theory attributes hypertension and hyperperfusion to endothelial damage, while a second theory suggests vasoconstriction and hypoperfusion as the trigger to generalized vasogenic edema of the cortex and subcortical white matter, leading to the described MR image [2–4]. Bevacizumab is a recombinant humanized monoclonal IgG antibody, which binds and inhibits vascular endothelial growth factors (VEGF) reducing angiogenesis and regressing tumor growth [3–5]. PRES has been associated with bevacizumab in multiple reported cases since 2006 [2–6]. The common features in these reports and other papers describing this syndrome are R. Sawaya (&) W. Radwan S. Hammoud American University Medical Center, POB 113-6044/C-27, Beirut, Lebanon e-mail: rs01@aub.edu.lb

Septin 9 promoter region methylation in free circulating DNA—potential role in noninvasive diagnosis of lung cancer: preliminary report

Abstract: Currently, there are no sensitive diagnostic tests that could allow early detection of lung cancer. Among some cancer patients, epigenetic changes in the nature of methylation of different gene promoter regions are observed, which affect expression of suppressor genes such as septin 9 (SEPT9). Due to the ability of detecting these changes in free circulating DNA in peripheral blood, such genes may become ideal markers in early and noninvasive diagnostics of cancer. Methylation of SEPT9 promoter region in plasma DNA is observed frequently in colorectal cancer patients. The aim of the study was to define the frequency of SEPT9 promoter methylation in lung cancer patients and evaluation of usefulness of this marker in early diagnostic of lung cancer. Plasma samples were obtained from 70 untreated patients with different lung cancer pathological diagnosis and disease stage and from 100 healthy individuals. DNA was isolated from peripheral blood plasma and was then subjected to bisulfitation, purification and elution using Abbott mSEPT9 Detection Kit. Methylation level was assessed by real-time PCR with the use of specific SEPT9 promoter methylation probe. Each sample was assayed in the presence of positive and negative control. SEPT9 promoter methylation was detected in 31 (44.3 % of the whole studied group) of lung cancer patients finding the result positive when methylation was detected in 1 out of 3 repetitions of each test sample determinations. The marker was present in patients with different pathological diagnosis and disease stage. Analysis of SEPT9 promoter region methylation may be useful in early diagnosis of lung cancer.

Elevated platelet to lymphocyte ratio predicts poor prognosis after hepatectomy for liver-only colorectal metastases, and it is superior to neutrophil to lymphocyte ratio as an adverse prognostic factor

Abstract: Recent evidence indicates that inflammation is an essential component of pathogenesis and progression of cancer. In this study, we analysed two indexes of systemic inflammation, the neutrophil-lymphocyte ratio (NLR) and platelet-lymphocyte ratio (PLR), with disease-free survival (DFS) and overall survival (OS) in liver-only colorectal metastases treated with liver resection following neoadjuvant chemotherapy. In this retrospective study, 140 patients were enroled. The NLR and PLR were calculated on the basis of preoperative blood cell count, and their cut-off levels were determined by applying receiver operating curve analysis. A NLR>2.4 and a PLR>150 were considered to be elevated. DFS and OS were calculated using both Kaplan-Meier and multivariate Cox regression methods. Both high NLR and high PLR were associated with decreased DFS [HR 1.55; 95% confidence interval (CI) 1.03-2.32; P=0.033, and HR 1.78; 95% CI 1.19-2.67; P=0.005, respectively] and OS (HR 2.21; 95% CI 1.24-3.96; P=0.007, and HR 2.90; 95% CI 1.61-5.21; P<0.001, respectively) in univariate analysis, but only PLR remained significant in multivariate analysis for both DFS and OS (HR 1.68; 95% CI 1.04-2.71; P=0.034, and HR 2.17; 95% CI 1.09-4.32; P=0.027, respectively). When we divided patients into three groups (group 1: normal both NLR and PLR; group 2: high NLR or high PLR; group 3: high both NLR and PLR), the five-year DFS and OS rates for these groups were 43, 26, 9% (P=0.004) and 73, 59, 34% (P<0.001), respectively. In this study, we indicate that preoperative PLR is superior to preoperative NLR as an adverse prognostic factor in patients who undergo liver resection for liver-only colorectal metastases.

Stemness state regulators SALL4 and SOX2 are involved in progression and invasiveness of esophageal squamous cell carcinoma

Abstract: Cancer stem cells, as a subgroup of tumor cells, resemble critical properties of embryonic stem cells (ESCs) such as self-renewal and maintenance of stemness state. SALL4 and SOX2 are two main transcription factors involving in maintenance of pluripotency, self-renewal and cell fate decision in ESCs. In this study, we aimed to elucidate the expression levels of these important transcription factors in esophageal squamous cell carcinoma (ESCC) and to reveal their probable roles in maintenance and progression of the disease. The expression level of SALL4 and SOX2 was analyzed in fresh tumoral tissues in comparison with distant tumor-free tissues of 50 ESCC patients by relative comparative real-time PCR. SALL4 and SOX2 were overexpressed in 64 and 32 % of tumor samples, respectively, in significant correlation with each other (p = 0.028). There was a significantly inverse correlation between low level of SALL4 expression and metastasis of tumor cells into the lymph nodes (p = 0.035). Furthermore, co-overexpression of the genes was significantly correlated with the depth of tumor invasion (p = 0.045) and metastasis to the lymph nodes (p = 0.049). SALL4 and SOX2 are co-overexpressed in ESCC and have a significant correlation with invasion and metastasis of the disease. To the best of our knowledge, this is the first report of SALL4 clinical relevance in ESCC to date. The clinical consequences of SALL4–SOX2 association suggest a possible functional interaction between these factors in regulation of ESCC maintenance and aggressiveness and introduce these regulators of stemness state as potentially interesting therapeutic targets to bring new opportunities for onco-therapeutic modalities.

Histone deacetylase inhibitors in glioblastoma: pre-clinical and clinical experience

Abstract: Epigenetic mechanisms are increasingly recognized as a major factor contributing to pathogenesis of cancer including glioblastoma, the most common and most malignant primary brain tumour in adults. Enzymatic modifications of histone proteins regulating gene expression are being exploited for therapeutic drug targeting. Over the last decade, numerous studies have shown promising results with histone deacetylase (HDAC) inhibitors in various malignancies. This article provides a brief overview of mechanism of anti-cancer effect and pharmacology of HDAC inhibitors and summarizes results from pre-clinical and clinical studies in glioblastoma. It analyses experience with HDAC inhibitors as single agents as well as in combination with targeted agents, cytotoxic chemotherapy and radiotherapy. Hallmark features of glioblastoma, such as uncontrolled cellular proliferation, invasion, angiogenesis and resistance to apoptosis, have been shown to be targeted by HDAC inhibitors in experiments with glioblastoma cell lines. Vorinostat is the most advanced HDAC inhibitor that entered clinical trials in glioblastoma, showing activity in recurrent disease. Multiple phase II trials with vorinostat in combination with targeted agents, temozolomide and radiotherapy are currently recruiting. While the results from pre-clinical studies are encouraging, early clinical trials showed only modest benefit and the value of HDAC inhibitors for clinical practice will need to be confirmed in larger prospective trials. Further research in epigenetic mechanisms driving glioblastoma pathogenesis and identification of molecular subtypes of glioblastoma is needed. This will hopefully lead to better selection of patients who will benefit from treatment with HDAC inhibitors.

High expression of miR-21 in triple-negative breast cancers was correlated with a poor prognosis and promoted tumor cell in vitro proliferation

Abstract: The triple-negative breast cancer (TNBC), with a particularly poor prognosis, is increasingly recognized as heterogeneous in molecular signatures. MicroRNA expression profiles have been used for the classification and prognostication of breast cancer, numerous significantly upregulated microRNAs, i.e. miR-21, have been verified oncogenic in non-TNBCs. In present study, we determined the miR-21 levels in TNBC specimens, and TNBC cell levels in vitro, and then identified the role of miR-21 on tumor cell proliferation, apoptosis, and then identified PTEN as the possible target of the microRNA. It was shown that miR-21 expression is upregulated generally, and heterogeneous in TNBC specimens, posing a correlation with poor prognosis for TNBC patients. Further results demonstrated that the upregulated miR-21 promoted the tumor proliferation and inhibited cell apoptosis in vitro. And pro-apoptotic PTEN had been shown being targeted and downregulated. Therefore, our finding emphasized the oncogenic role of miR-21 in TNBC.

Sonic hedgehog signaling may promote invasion and metastasis of oral squamous cell carcinoma by activating MMP-9 and E-cadherin expression

Abstract: The aim of this study was to investigate sonic hedgehog (SHH) signaling pathway components (Shh and Gli-1), E-cadherin, and MMP-9 expression in human oral squamous cell carcinoma (OSCC) and to evaluate their role in prognosis. Expression of Shh, Gli-1, and MMP-9 was significantly upregulated in 74 OSCC samples compared with non-cancerous tissue samples (Shh IOD: 162.44 ± 29.35 and 608.82 ± 170.99; Gli-1 IOD: 203.50 ± 71.57 and 831.11 ± 242.352; MMP-9 IOD: 196.69 ± 64.48 and 721.64 ± 197.99 in non-cancerous and tumor tissues, respectively, P < 0.01), whereas E-cadherin expression was downregulated (E-cadherin IOD: 1,006.19 ± 230.42 and 442.20 ± 156.11; in non-cancerous and tumor tissues, respectively, P < 0.01). Highly expressed proteins were associated with lymph node metastasis; moreover, overexpression of Gli-1 was related to tumor recurrence and cancer clinical staging. Spearman’s analysis indicated that the expression of Gli-1 and MMP-9 was positively correlated, whereas expression of Shh/Gli-1 and E-cadherin was negatively correlated. Kaplan–Meier results revealed that patients with low Shh, Gli-1, and MMP-9 expression survived longer than those with high expression. In contrast, low E-cadherin expression was associated with poor prognosis (P < 0.01). In conclusions, transcription factor Gli-1 of the SHH signaling pathway may be an important mediator of invasion and metastasis of OSCC through induced expression of MMP-9 and E-cadherin and may serve as a new prognostic marker.

Cancer stem cell detection and isolation

Abstract: Only 10 % of cancer-related deaths result from primary tumors; most are caused by metastatic tumors. It is believed that the metastatic power of tumor cells is attributed to features of a stem cell-like subpopulation of tumor cells known as cancer stem cells (CSCs). Cancer stem cells are resistant to chemotherapeutic treatments and can induce dormancy in tumor cells for long periods. Detection, isolation, and characterization of CSCs in solid tumors are hallmarks of cancer-targeted therapies in recent years. There are inevitable similarities between normal and cancer stem cells; therefore, finding specific methods or markers to differentiate them is critical to cancer therapies. Considering CSCs involvement in tumor relapse and chemotherapeutic resistance, identification of such cells in tumors is imperative for effective targeted therapy. The present review introduces practical and specific protocols used to isolate CSCs from solid tumors from colon, esophagus, liver, breast, brain, and cervix.

miR-99a and -99b inhibit cervical cancer cell proliferation and invasion by targeting mTOR signaling pathway

Abstract: MicroRNAs were demonstrated to play an important role in the regulation of gene expression. Here, we showed that miR-99a and -99b (miR-99a/b) were down-regulated in human cervical cancer patient tissues and were negatively related with lymphatic metastasis. In addition, overexpression of miR-99a/b inhibited cell growth and invasion, whereas suppression of miR-99a/b yielded the reverse phenotype. Dual luciferase report assay revealed that mTOR was identified as a novel target gene of both miR-99a and -99b. Altogether, these results suggested that miR-99a/b directly and negatively regulated mTOR expression in cervical cancer cells, and enforced the importance of miR-99a/b and their targets in the malignant phenotypes of cervical carcinogenesis.

miR-139 targets CXCR4 and inhibits the proliferation and metastasis of laryngeal squamous carcinoma cells

Abstract: Our previous studies have showed that chemokine receptor 4 (CXCR4) was over-expressed in laryngeal squamous cell carcinoma (LSCC). However, the mechanism underlying aberrant CXCR4 expression remains unclear. To investigate the roles played by miRNAs in CXCR4 over-expression in LSCC, putative miR-139 was predicted through computational algorithms, including TargetScan, PicTar and miRBase, and luciferase reporter assay was explored to confirm that whether CXCR4 was directly regulated by miR-139. Then, quantitative real-time PCR, immunohistochemistry and in situ hybridization methods were employed to detect the expression of miR-139 and CXCR4 in primary LSCC tissues, normal adjacent mucosal tissues and metastatic lesions derived from 40 LSCC patients in the Second Hospital, Xi'An JiaoTong University. Finally, gain- and loss-of-function assays were adopted to explore the effects of miR-139 and CXCR4 on proliferation, invasion and metastasis of the human LSCC cell line Hep-2 in vitro and in vivo. Our results showed that miR-139 dampened CXCR4 expression, and CXCR4 was directly targeted by miR-139. Additionally, the expression of miR-139 was reduced in alignment with the progression of primary to metastatic LSCC. Moreover, an inverse correlation was observed between miR-139 and CXCR4 protein levels in LSCC specimens. Functional analyses demonstrated that ectopic expression of miR-139 inhibited cell proliferation, migration and metastasis of Hep-2 cells in vitro and in vivo. Similar to the observations seen in restoring miR-139 expression, dampening of CXCR4 expression inhibited cell growth, migration and invasion, whereas miR-139 over-expression reversed the pro-metastatic effect of CXCR4. Taken together, we conclude that miR-139 targets CXCR4 and inhibits proliferation and metastasis of LSCC.

Clinical significance of HuR expression in human malignancy

Abstract: Hu-antigen R (HuR) is an RNA-binding protein that regulates the stability, translation, and nucleus-to-cytoplasm translocation of target mRNAs. The aim of the present review was to summarize and present the currently available information in the English literature on HuR expression in various human tumors, verifying its possible clinical significance. HuR function is directly linked to its subcellular localization. In normal cells, HuR is mostly localized in the nucleus, while in malignant cells, an increase in cytoplasmic HuR levels has been noted, in both cell lines and tissue samples. Moreover, in malignancy, elevated HuR expression levels and cytoplasmic immunohistochemical pattern have been correlated with advanced clinicopathological parameters and altered expression levels of proteins implicated in neoplasia. Additionally, elevated HuR expression levels and mainly cytoplasmic immunohistochemical pattern were correlated with decreased patients’ survival rate in various human tumors. HuR is a putative drug target for cancer therapy, since it is expressed ubiquitously in malignant clinical samples and has an apparently consistent role in tumor formation and progression.

Predicting distant metastasis and chemoresistance using plasma miRNAs.

Abstract: In the clinic, predicting metastasis and chemoresistance takes high priority, but has not been well established. This study seeks to investigate whether dynamically monitoring serum microRNAs (miRNAs) can help predict metastasis, chemoresistance, and prognosis of colorectal cancer. Serum miR-155, miR-200c, and miR-210 levels in 15 patients with colon cancer were measured by real-time PCR at different time points post surgery and chemotherapy for 3 years. Significant increases in miR-155, miR-200c, and miR-210 levels were observed in the serum and tumor tissues of colon cancer patients compared to that of healthy subjects. After surgery and chemotherapy, the serum levels of these miRNAs in patients with good prognosis returned to normal levels found in healthy controls during the 3-year follow-up. In patients with recurrence and distant metastasis, serum miR-155, miR-200c, and miR-210 levels remained at an elevated level or became elevated again after a short period of decline. In patients with good response to chemotherapy for metastatic tumors, re-elevation of miR-155 was not significant compared to miR-200c and miR-210. In contrast, miR-155 re-elevated more significantly in patients not sensitized to chemotherapy than miR-200c and miR-210. Our study suggests that re-elevation or sustained elevation of serum miR-155 level after surgery and chemotherapy is a sign of chemoresistance in colon cancer, while high and/or re-elevated miR-155, miR-200c, and miR-210 levels implicate local recurrence and distant metastasis as well as poor prognosis.

Low expression of novel lncRNA RP11-462C24.1 suggests a biomarker of poor prognosis in colorectal cancer.

Abstract: Long noncoding RNAs (lncRNAs) have recently emerged as a major class of regulatory molecules, which were involved in a broad range of biological processes and complex diseases. Research on lncRNAs may shed light on tumorigenesis and progression of colorectal cancer (CRC). The purpose of the present study was to identify lncRNAs correlated with CRC and then investigate their potential functions. We selected 92 patients for this prospective study and then collected the tumor samples and clinical records. First, the global lncRNA expression profiles in tumor and adjacent normal tissues of patients with non-metastatic CRC and patients with metastatic CRC were measured by microarray assay. Then, a noteworthy lncRNAs RP11-462C24.1 whose function was previously unknown was explored in detail on the aspect of the association of its expression level and clinicopathological features of CRC and patients’ survival. We found that RP11-462C24.1 expression level was lower in cancer tissues compared with adjacent normal samples (P < 0.001). Furthermore, its expression level was lower in CRC patients with metastasis than those without metastasis (P = 0.049). That is, RP11-462C24.1 expression level decreased as the malignant degree of CRC increased. In addition, low expression of RP11-462C24.1 significantly correlated with more distant metastasis (P = 0.011). The areas under ROC curves were 0.78 and 0.65 for RP11-462C24.1, distinguishing CRC from normal tissue and distinguishing CRC without metastasis from CRC with metastasis, respectively. Multivariate analysis identified that RP11-462C24.1 was an independent predictor for patients prognosis (P = 0.005). Furthermore, Kaplan–Meier analysis showed that patients with low expression of RP11-462C24.1 had a poor disease-free survival (P < 0.001). This is the first study that correlates RP11-462C24.1 expression profile with malignancy grade in human CRC. Our results showed that RP11-462C24.1 could be a potential novel prognostic marker for CRC, and thus, provided a new strategy for CRC diagnosis. Meanwhile, our findings indicated the potential roles of RP11-462C24.1 in tumorigenesis and progression of CRC, which gave a clue for future studies.

MicroRNA-335 inhibits invasion and metastasis of colorectal cancer by targeting ZEB2

Abstract: MicroRNAs have been suggested to play a vital role in regulate tumor progression and invasion. However, the expression of miR-335 in colorectal cancer (CRC) and its clinical significance are not known. Here, we report that miR-335 is a tumor suppressor by regulating expression of ZEB2. In this study, we showed that downregulated miR-335 levels in highly invasive CRC cell lines and tissues. Kaplan–Meier survival analysis indicated that patients with reduced miR-335 had a poor overall survival. Furthermore, enhancing the expression of miR-335 inhibited CRC cell migration and invasion in vitro and lung and liver metastasis in vivo, while silencing its expression resulted in increased migration and invasion. Additionally, we identified a novel miR-335 target, ZEB2, and the direct interaction between them was verified by 3′-untranslated region dual-luciferase reporter assay. In conclusion, our results demonstrate that miR-335 functions as a tumor suppressor and play a role in inhibiting metastasis of CRC cells through targeting ZEB2. These findings suggest that miR-335 may be useful as a new potential therapeutic target for CRC.

Three dysregulated microRNAs in serum as novel biomarkers for gastric cancer screening

Abstract: Gastric cancer (GC) is one of the most threatening diseases. The symptoms of GC are complex and hard to detect, which also contribute to the poor prognosis of GC. Besides, the current diagnosis for GC is expensive and invasive. Thus, a fast, noninvasive biomarker is urgently needed for GC screening. MicroRNAs (miRNAs) are small noncoding RNAs, which are involved in a great variety of pathological processes, particularly carcinogenesis. MiRNAs are stable in gastric juice, plasma as well as serum, which facilitate it to be a promising biomarker for cancer. In this study, we selected three novel miRNAs, i.e., miR-233, miR-16, and miR-100, to investigate their potential diagnostic value in GC screening. A total of 50 GC patients and 47 healthy controls were involved in this study. Blood serum samples were collected; RNAs were extracted and normalized with U6 snRNA as the internal control; qRT-PCR was performed for relative expression of target miRNAs. Levels of miRNAs expression were compared by Student’s t test for the comparison between two groups, and one-way ANOVA was used for multiple comparisons. The expression of miR-223, miR-16, and miR-100 was all significantly higher in GC patients than controls (all P < 0.001). All the tested miRNAs were manifested to be valuable biomarkers for GC. Relative expression of these miRNAs was significantly correlated with clinical characteristics of GC patients, such as TNM stage (P = 0.036 for miR-223; P < 0.001 for miR-100), metastatic status (P = 0.045 for miR-223; P = 0.031 for miR-16; P = 0.006 for miR-100), tumor size (P = 0.042 for miR-223; P = 0.031 for miR-16; P < 0.001 for miR-100), and differentiation grade (P = 0.036 for miR-223; P = 0.030 for miR-16; P = 0.034 for miR-100). However, in T classification, which considered both tumor size and direct extent of primary tumor, the difference in target miRNAs expression was not significant. In summary, we confirmed the diagnostic value of serum miR-223, miR-16, and miR-100 in GC. Significantly elevated expression of the three miRNAs was also observed in advanced GC patients, which suggested their availability in cancer staging.

Prognostic impact of circulating monocytes and lymphocyte-to-monocyte ratio on previously untreated metastatic non-small cell lung cancer patients receiving platinum-based doublet

Abstract: The link between circulating lymphocyte-to-monocyte ratio (LMR) and newly diagnosed metastatic non-small cell lung cancer (NSCLC) is not fully defined. The study was conducted to evaluate the prognostic impact of LMR on survival outcomes in previously untreated metastatic NSCLC patients receiving platinum-based doublet. Chemotherapy-naive metastatic NSCLC patients undergoing platinum-based doublet were retrospectively enrolled. Clinical features regarding gender, age, Eastern Cooperative Oncology Group (ECOG) performance status, histology, absolute lymphocyte count (ALC), absolute monocyte count (AMC) and LMR were collected to determinate their prognostic impact on progression-free survival (PFS) and overall survival (OS). Up to 370 patients were eligible for the study. By univariate analysis, ECOG performance status, histology, ALC, AMC and LMR were showed to be significantly associated with PFS and OS. In subsequent Cox multivariate analysis, non-squamous cell carcinoma, ALC ≥ 2.45 × 10(9)/L, AMC <0.45 × 10(9)/L and LMR ≥ 4.56 were demonstrated to be independently correlated with better PFS. In addition, independent favorable prognostic factors for OS were only limited to LMR ≥ 4.56 and non-squamous cell carcinoma, whereas ECOG performance status of 2 and AMC ≥ 0.45 × 10(9)/L remained as independently inferior prognostic indicators for OS. Our findings implicate that circulating AMC and LMR are regarded as independent prognostic factors for PFS and OS in previously untreated metastatic NSCLC patients receiving platinum-based doublet.

Discordances in ER, PR and HER2 receptors between primary and recurrent/metastatic lesions and their impact on survival in breast cancer patients

Abstract: The aim of this study was to evaluate the frequency and prognostic impact of changes in the estrogen (ER), progesterone (PR) and human epidermal growth factor receptor 2 (HER2) status between primary and recurrent/metastatic lesions (RML). We investigated 133 breast cancer patients for ER, PR and HER2 status of primary and RML and their follow-up records. Among 133 patients with RML, discordance rate for ER, PR, and HER2 was 18.8, 33.8, and 6.8%, respectively. ER, PR and HER2 discordance were observed in 20.0, 38.1 and 6.7% of the patients with distant metastasis, and in 14.3, 17.9 and 7.1% of the patients with locoregional recurrence. The mean time between the primary diagnosis and last contact or death was 57 (range 22-78) months and between the recurrence biopsy and last contact or death was 17 (range 1-33) months. Among 133 patients with RML, the ER-discordant cases and ER-loss cases experienced a worse overall survival (OS) (p=0.001 and p=0.016, respectively) and post-recurrence survival (PRS) (p=0.001 and p=0.018, respectively), compared with the respective concordant cases. The HER2-discordant patients and HER2-loss patients had a poorer OS (p=0.008 and p=0.001, respectively) and PRS (p=0.004 and p=0.000, respectively) than the respective concordant cases. Among 105 patients with distant metastasis, ER discordance, ER loss, HER2 discordance and HER2 loss, compared with the respective concordant cases, resulted in a worse OS and PRS (p<0.05 for all). Our findings show an evident change in ER, PR and HER2 between breast primary tumors and relapsing tumors. The unstable status for ER or HER2 in breast cancer seems to be clinically significant and to correlate with a worse prognosis.

Downregulation of E-cadherin expression in breast cancer by promoter hypermethylation and its relation with progression and prognosis of tumor

Abstract: Breast cancer is the most common cancer in women around the world, and novel prognosis strategies is needed to control more accurate and effective of this malignant disease. Among the latest prognostic markers is E-cadherin, which mediates cell–cell adhesion by associating with catenins. Loss of E-cadherin gene (CDH1) function by genetic or epigenetic alteration leads to tumorigenesis. The aim of our study was to investigate E-cadherin gene promoter methylation in breast cancer, and its correlation with E-cadherin protein expression. Fifty primary breast cancers tissue with ductal type and 50 normal breast sample from the same patients that was located adjacent to tumor region as controls were provided by Imam Reza-based referral and teaching hospital affiliated to Tabriz University of Medical Sciences, Tabriz, Iran. CDH1 promoter region CpG sites methylation and E-cadherin protein expression were determined by bisulfite-specific polymerase chain reaction and Western blot analysis, and the resulting products were sequenced on an ABI automated sequencer for firm conclusion. CDH1 hypermethylation in breast tumor specimen (ductal type) was observed in 94 % (47 of 50) comparing with normal samples methylation, and the significant difference was (p = 0.000). Protein expression in tumor samples tends to diminish with the CDH1 promoter region methylation. In the group of 50 ductal carcinomas cases, most of the cases showing CDH1 hypermethylation correlated inversely with the reduced levels of expression of E-cadherin proteins (95 % of full-methylated tumor samples had no protein expression, and 4.5 % of them had weak expression levels). Possible association was observed between CDH1 methylation and its protein expression (p = 0.000). The results of methylation analysis in promoter region in ten CpG sites (863, 865, 873, 879, 887, 892, 901, 918, 920, and 940) suggested that abnormal CDH1 methylation occurs in high frequencies in ductal breast tumors probably sounds the process of carcinogenesis progression.

Over-expression of survivin and VEGF in small-cell lung cancer may predict the poorer prognosis

Abstract: The expression of survivin, an inhibitor of apoptosis can be seen in most tumors and is correlated with the angiogenic factor vascular endothelial growth factor (VEGF). But little is known about their contribution in small-cell lung cancer (SCLC). This study was designed to investigate the expression of survivin and VEGF in SCLC, and to explore their correlation with clinical-pathological feature and prognosis. Forty-five patients with pathological histology of SCLC were entered into this study. Forty-five cases of matched adjacent non-tumor samples and 10 samples of operated patients with benign lung tumor were also included as control. The expression of survivin and VEGF was detected by immunohistochemistry (IHC, SP). These two sets of data were processed and tested for correlation with major patients' characteristics, and overall survival. The correlations between survivin and VEGF expressions and the clinical-pathological features were evaluated by chi-square test. The correlation between survivin and VEGF expressions was analyzed by Spearman's rank correlation test; the overall survival was analyzed by the Kaplan-Meier method; and the relationship between clinical and pathological features and overall survival was analyzed by the Cox proportional hazard models. Positive expression rate of survivin and VEGF was significantly higher in SCLC than those of adjacent non-tumor tissues and benign lung tumor tissues (73.3 vs. 15.6 vs. 0 %, P < 0.05) and (75.6 vs. 20 vs. 0 %, P < 0.05), respectively. Survivin and VEGF expressions were significantly associated with lymph node metastasis (P = 0.003, 0.011) and clinical stage (P = 0.006, 0.021). The expression of survivin was significantly coincident with the expression of VEGF (r = 0.644, P = 0.000). The median overall survival in survivin positive group and VEGF positive group was significantly shorter than those in survivin negative and VEGF negative group, respectively (log-rank P = 0.000). Moreover, multivariate analysis showed that survivin expression (HR 0.224; 95 % CI 0.074-0.675; P = 0.008) and VEGF expression (HR 0.172; 95 % CI 0.054-0.559; P = 0.003) were statistically independent predictive factors of poorer prognosis for SCLC patients. Our results indicated that survivin and VEGF were over-expressed in small-cell lung cancer, each of them may be an independent poor prognostic factor.