Plasma miRNAs in diagnosis and prognosis of pancreatic cancer: A miRNA expression analysis.

Lung cancer is the leading cause of cancer death in the world, and aberrant expression of miRNA is a common feature during the cancer initiation and development. Our previous study showed that levels of miRNA-148a assessed by quantitative real-time polymerase chain reaction (qRT-PCR) were a good prognosis factor for non-small cell lung cancer (NSCLC) patients. In this study, we used high-throughput formalin-fixed and paraffin-embedded (FFPE) lung cancer tissue arrays and in situ hybridization (ISH) to determine the clinical significances of miRNA-148a and aimed to find novel target of miRNA-148a in lung cancer. Our results showed that there were 86 of 159 patients with low miRNA-148a expression and miRNA-148a was significantly down-regulated in primary cancer tissues when compared with their adjacent normal lung tissues. Low expression of miRNA-148a was strongly associated with high tumor grade, lymph node (LN) metastasis and a higher risk of tumor-related death in NSCLC. Lentivirus mediated overexpression of miRNA-148a inhibited migration and invasion of A549 and H1299 lung cancer cells. Furthermore, we validated Wnt1 as a direct target of miRNA-148a. Our data showed that the Wnt1 expression was negatively correlated with the expression of miRNA-148a in both primary cancer tissues and their corresponding adjacent normal lung tissues. In addition, overexpression of miRNA-148a inhibited Wnt1 protein expression in cancer cells. And knocking down of Wnt-1 by siRNA had the similar effect of miRNA-148a overexpression on cell migration and invasion in lung cancer cells. In conclusion, our results suggest that miRNA-148a inhibited cell migration and invasion through targeting Wnt1 and this might provide a new insight into the molecular mechanisms of lung cancer metastasis.

/pdf/mirna-148a-serves-as-a-prognostic-factor-and-suppresses-57mcelducz.pdf

miRNA-148a serves as a prognostic factor and suppresses migration and invasion through Wnt1 in non-small cell lung cancer

Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease with a dismal prognosis for which new therapeutic strategies are desperately needed. Non-coding RNAs (ncRNAs), especially microRNAs (miRNAs) and long non-coding RNAs (lncRNAs), may yield new therapeutic concepts for the treatment of PDAC. A vast number of miRNAs, including the well-studied miR-21, miR-155 and miR-34, has been shown to regulate PDAC growth, invasion and metastasis in vitro and in vivo by targeting members of key signaling pathways. In addition, other miRNAs and lncRNAs, such as HOTTIP and MALAT-1, have been shown to influence the malignant behavior of PDAC cells. Here, we discuss several ncRNAs that may be used either as new therapeutic agents or as targets of new therapeutic agents. Furthermore, we discuss the problem of proper delivery of nucleotide-based agents and novel methods that may be used to circumvent this problem. Although the number of reports addressing the role of ncRNAs in PDAC virtually grows by the day, there are still many steps to be taken before the application of ncRNA-based therapies will become reality in clinical practice.

Non-coding RNAs in pancreatic cancer: challenges and opportunities for clinical application.

El cancer de seno es el cancer mas comun y tambien es la primera causa de muerte por cancer en mujeres a nivel mundial. En Colombia es la primera causa de muerte por cancer en mujeres, de ahi la importancia en la identificacion de genes que puedan estar implicados en el desarrollo y progresion de la enfermedad. En este trabajo se llevo a cabo un estudio de asociacion tipo gen candidato en el gen DEAR1. El gen DEAR1, codifica para un miembro de la subfamilia TRIM de proteinas “finger RING” (TRIM 62), esta ubicado dentro del cromosoma 1p35.1, presenta mutaciones y deleciones homocigotas en cancer de seno y su expresion esta disminuida o perdida en DCIS (carcinoma ductal in situ). DEAR1 es una E3 ubiquitin ligasa, involucrada en el proceso de morfogenesis acinar de la glandula mamaria. Es un supresor tumoral que actua como un regulador dominante de polaridad, arquitectura del tejido (Lott ST et al., 2009) y como un regulador del proceso de transicion epitelio mesenquimal dirigido por TGFβ (Chen N et al., 2013). El objetivo principal del estudio fue determinar la asociacion de variantes moleculares en el gen DEAR1 con cancer de seno, parametros histopatologicos y sobrevida usando una aproximacion “tagging SNP” en una muestra de poblacion Colombiana. Se realizo un estudio caso – control que incluyo 1023 pacientes afectadas con cancer de seno (casos) y 1023 pacientes sin la enfermedad (controles), se incluyeron datos epidemiologicos, histopatologicos y de sobrevida. Se realizo la genotipificacion de cuatro SNPs rs584298, rs2927970, rs59983645 y rs599167 que capturaron el 93,8% de la variacion genetica en la region investigada considerando un MAF≥5% y r2≥0,8, para su genotipificacion se utilizaron dos aproximaciones metodologicas iPLEX y KASP. Se realizo el analisis de asociacion por regresion condicional logistica para seis variables de riesgo epidemiologico y variables patologicas. Los resultados revelaron una asociacion entre el tagSNP rs584298 genotipos AG y GG con cancer de seno [p=0.048 y p=0.004 respectivamente] y el rs584298 (genotipo AG p=0.015) mostro asociacion con la expresion de PR y el genotipo AA (p=0.048) mostro asociacion con la expresion de HER2. La secuenciacion de la region codificante del gen DEAR1 en tejidos tumorales de cancer de seno y tejidos normales embebidos en parafina fue realizada con el fin de identificar nuevas variantes. Los analisis de secuencia realizados con el software CLC genomics workbench revelaron una baja longitud de distribucion de las lecturas (1000pb). En el analisis se evidencio la presencia de una variante ya anotada en el NCBI rs622407 que se localiza en el dominio RING del gen TRIM62, es un SNP sinonimo que no altera la funcionalidad de la proteina. Estos resultados sugieren que variantes en DEAR1 estan asociadas con cancer de seno en una muestra de la poblacion colombiana lo cual se traduciria en el posible uso de estas variantes como factores pronosticos de la enfermedad.

Asociación de variantes genéticas en el gen DEAR1 con cáncer de seno y desenlace clínico en población colombiana

Circular RNAs (circRNAs) have been reported to participate in the molecular mechanism of human cancers. The PUM1 gene has been confirmed to be closely related to tumorigenesis and progression of ovarian cancer. In the present study, we explored the function and underlying molecular mechanism of circPUM1 in ovarian cancer. qRT-PCR analysis showed upregulation of circPUM1 in ovarian cancer tissues compared with normal ovaries. Gain- and loss-of-function experiments indicated that circPUM1 increased cell proliferation, migration, and invasion and inhibited cell apoptosis. Intraperitoneal injection of circPUM1-knockout tumor cells in nude mice resulted in a decrease in the metastatic ability of the tumor. Bioinformatics analysis and dual-luciferase reporter assays revealed that circPUM1 upregulated the expression of nuclear factor kappa B (NF-κB) and MMP2 by sponging miR-615-5p and miR-6753-5p. Further studies showed that exosomal circPUM1 acted on peritoneal mesothelial cells and increased tumor metastasis. In conclusion, our study indicates that circPUM1 not only promotes ovarian cancer proliferation, migration and invasion, but also acts on the peritoneum and contributes to metastasis of cancer in the form of cancer-derived exosomes.

circPUM1 Promotes Tumorigenesis and Progression of Ovarian Cancer by Sponging miR-615-5p and miR-6753-5p

Modulation of KRAS activity by upstream signals has revealed a promising new approach for pancreatic cancer therapy; however, it is not clear whether microRNA-associated KRAS axis is involved in the carcinogenesis of pancreatic cancer. Here, we identified miR-193b as a tumor-suppressive miRNA in pancreatic ductal adenocarcinoma (PDAC). Expression analyses revealed that miR-193b was downregulated in (10/11) PDAC specimens and cell lines. Moreover, we found that miR-193b functioned as a cell-cycle brake in PDAC cells by inducing G1-phase arrest and reducing the fraction of cells in S phase, thereby leading to dampened cell proliferation. miR-193b also modulated the malignant transformation phenotype of PDAC cells by suppressing anchorage-independent growth. Mechanistically, KRAS was verified as a direct effector of miR-193b, through which the AKT and ERK pathways were modulated and cell growth of PDAC cells was suppressed. Taken together, our findings indicate that miR-193b-mediated deregulation of the KRAS axis is involved in pancreatic carcinogenesis, and suggest that miR-193b could be a potentially effective target for PDAC therapy.

/pdf/deregulation-of-the-mir-193b-kras-axis-contributes-to-3qszekgkb2.pdf

Deregulation of the MiR-193b-KRAS Axis Contributes to Impaired Cell Growth in Pancreatic Cancer

Background 
Aberrant microRNA (miRNA) expression is associated with tumor development. This study aimed to elucidate the role of miR-615-5p in the development of pancreatic ductal adenocarcinoma (PDAC).


Methods 
Locked nucleic acid in situ hybridization (LNA-ISH) was performed to compare miR-615-5p expression in patients between PDAC and matched adjacent normal tissues. Effects of miR-615-5p overexpression on cell proliferation, apoptosis, colony formation, migration, and invasion were determined in the pancreatic cancer cell lines PANC-1 and MIA PaCa-2. Effects of miR-615-5p on AKT2 were examined by dual-luciferase reporter assay. Lentivirus expressing miR-615 was used to create stable overexpression cell lines, which were subsequently used in mouse xenograft and metastasis models to assess tumor growth, apoptosis and metastasis.


Results 
miR-615-5p expression was significantly lower in PDAC than in adjacent normal tissues. Low levels of miR-615-5p were independently associated with poor prognosis (HR: 2.243, 95% CI: 1.190-4.227, P=0.013). AKT2 protein expression was inversely correlated with miR-615-5p expression (r=-0.3, P=0.003). miR-615-5p directly targeted the 3’-untranslated region of AKT2 mRNA and repressed its expression. miR-615-5p overexpression inhibited pancreatic cancer cell proliferation, migration, and invasion in vitro, and tumor growth and metastasis in vivo. Furthermore, miR-615-5p overexpression also induced pancreatic cancer cell apoptosis both in vitro and in vivo.


Conclusions 
These results show that miR-615-5p inhibits pancreatic cancer cell proliferation, migration, and invasion by targeting AKT2. The data implicate miR-615-5p in the prognosis and treatment of PDAC.

/pdf/mirna-615-5p-functions-as-a-tumor-suppressor-in-pancreatic-3kat2dw0r8.pdf

MiRNA-615-5p Functions as a Tumor Suppressor in Pancreatic Ductal Adenocarcinoma by Targeting AKT2

Objective To investigate the role of miR-150-5p in cell proliferation and apoptosis in human pancreatic cancer cell lines.Methods The expression of miR-150-5p in pancreatic cancer was detected by real time qPCR analysis in 11 pairs of pancreatic cancer tissue and matched adjacent normal tissue samples and in 4 pancreatic cancer cell lines.PANC-1,MIA PaCa-2,BxPC-3 and AsPC-1 cells were transfected with chemically synthesized MiR-150-5p mimics,and CCK-8 assays was then performed to assess cellular functions.To fully understand the mechanisms by which miR-150-5p exerted its function,cell cycle analysis was performed on MIA PaCa-2 and PANC-1 cells 48 hours after transfection,by incubating with propidium iodide (PI)and subsequently analyzed by fluorescence-activated cell sorting (FACS).Apoptosis assay was performed on MIA PaCa-2 and PANC-1 cell lines 24 hours after transfection using the Annexin V-FITC Apoptosis Detection Kit I (BD Biosciences) and analyzed by FACS.Results The expression of miR-150-5p was consistently lower in the pancreatic cancer tissues than in normal tissues,and the miR-150-5p was also down-regulated in pancreatic cancer cell lines (P ＜ 0.05).MiR-150-5p mimics transfection siguificantly raised the expression level of miR-150-5p mRNA in PANC-1 and MIA PaCa-2 (P ＜0.01).The CCK-8 proliferation assay showed that cell growth was reduced in 4 pancreatic cancer cell lines (AsPC-1,BxPC-3,MIA PaCa-2,PANC-1) of miR-150-5p transfected cells compared with NC-transfected cells.The inhibition rates were 50.7％,48.6％,30.8％ and 42.3％,respectively (P ＜ 0.01).The apoptotic rate was increased in cells transfected with miR-150-5p mimics (P ＜0.01).The cell cycle analysis in MIA PaCa-2 indicated that miR-150-5p treatment induced cell cycle arrest in G1 phase with a significant increase in the percentage of cells in G1 phase (P ＜ 0.01),and a reduction of the S-phase cell population in MIA PaCa-2 and PANC-1 (P＜0.01).Conclusions MiR-150-5p is down-regulated in pancreatic cancer.Overexpression of miR-150-5p inhibits cell proliferation,blocked the cell cycle,but promotes cell apoptosis in pancreatic cancer cells.

Key words: 
Pancreatic neoplasms ;  MicroRNAs;  Tumor cells,cultured

[MiR-150-5p inhibits the proliferation and promoted apoptosis of pancreatic cancer cells].

There is an error in Table 2. The values for “Negative,” “Weak positive,” and “Positive rate” in the “PDAC” column are incorrect. Please see the corrected Table 2 here.



Table 2

miR-615-5p expression in PDAC and normal pancreas tissues determined by locked nucleic acid in situ hybridization (LNA-ISH)

/pdf/correction-mirna-615-5p-functions-as-a-tumor-suppressor-in-2jps6dyvei.pdf

Xianglan Jin

Papers

Deregulation of the MiR-193b-KRAS Axis Contributes to Impaired Cell Growth in Pancreatic Cancer

MiRNA-615-5p Functions as a Tumor Suppressor in Pancreatic Ductal Adenocarcinoma by Targeting AKT2

[MiR-150-5p inhibits the proliferation and promoted apoptosis of pancreatic cancer cells].

Correction: MiRNA-615-5p Functions as a Tumor Suppressor in Pancreatic Ductal Adenocarcinoma by Targeting AKT2