Hepatocellular carcinoma (HCC) is a complex biological process and is often diagnosed at advanced stages with no effective treatment options. With advances in tumor biology and molecular genetic profiling, several different signaling pathways and molecular mechanisms have been identified as responsible for initiating and promoting HCC. Targeting these critical pathways, which include the receptor tyrosine kinase pathways, the Ras mitogen-activated protein kinase (Ras/Raf/MAPK), the phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR), the Wnt/β-catenin signaling pathway, the ubiquitin/proteasome degradation and the hedgehog signaling pathway has led to the identification of novel therapeutics for HCC treatment. In this review, we elaborated on our current understanding of the signaling pathways involved in the development and initiation of HCC and anticipate the potential targets for therapeutic drug development.

https://www.mdpi.com/2072-6694/12/2/491/pdf

Molecular Signaling Pathways and Therapeutic Targets in Hepatocellular Carcinoma.

Macrophage-mimic shape changeable nanomedicine retained in tumor for multimodal therapy of breast cancer

The rapid development of CRISPR/Cas9 systems has opened up tantalizing prospects to sensitize cancers to chemotherapy using efficient targeted genome editing, but safety concerns and possible off-target effects of viral vectors remain a major obstacle for clinical application. Thus, the construction of novel nonviral tumor-targeting nanodelivery systems has great potential for the safe application of CRISPR/Cas9 systems for gene-chemo-combination therapy. Here, we report a polyamidoamine-aptamer-coated hollow mesoporous silica nanoparticle for the co-delivery of sorafenib and CRISPR/Cas9. The core-shell nanoparticles had good stability, enabled ultrahigh drug loading, targeted delivery, and controlled-release of the gene-drug combination. The nanocomplex showed >60% EGFR-editing efficiency without off-target effects in all nine similar sites, regulating the EGFR-PI3K-Akt pathway to inhibit angiogenesis, and exhibited a synergistic effect on cell proliferation. Importantly, the co-delivery nanosystem achieved efficient EGFR gene therapy and caused 85% tumor inhibition in a mouse model. Furthermore, the nanocomplex showed high accumulation at the tumor site in vivo and exhibited good safety with no damage to major organs. Due to these properties, the nanocomplex provides a versatile delivery approach for efficient co-loading of gene-drug combinations, allowing for precise gene editing and synergistic inhibition of tumor growth without apparent side effects on normal tissues.

Co-delivery of Sorafenib and CRISPR/Cas9 Based on Targeted Core–Shell Hollow Mesoporous Organosilica Nanoparticles for Synergistic HCC Therapy

Cancer is a leading cause of morbidity and mortality worldwide. Apoptosis is a process of programmed cell death and it plays a vital role in human development and tissue homeostasis. Mounting evidence indicates that apoptosis is closely related to the survival of cancer and it has emerged as a key target for the discovery and development of novel anticancer drugs. Various studies indicate that targeting the apoptotic signaling pathway by anticancer drugs is an important mechanism in cancer therapy. Therefore, numerous novel anticancer agents have been discovered and developed from traditional Chinese medicines (TCMs) by targeting the cellular apoptotic pathway of cancer cells and shown clinically beneficial effects in cancer therapy. This review aims to provide a comprehensive discussion for the role, pharmacology, related biology, and possible mechanism(s) of a number of important anticancer TCMs and their derivatives mainly targeting the cellular apoptotic pathway. It may have important clinical implications in cancer therapy.

/pdf/apoptotic-pathway-as-the-therapeutic-target-for-anticancer-t5r3q1sr43.pdf

Apoptotic Pathway as the Therapeutic Target for Anticancer Traditional Chinese Medicines.

Hepatocellular carcinoma (HCC) is the most common type of liver cancer and one of the leading causes of cancer-related death worldwide. Advanced HCC displays strong resistance to chemotherapy, and traditional chemotherapy drugs do not achieve satisfactory therapeutic efficacy. Sorafenib is an oral kinase inhibitor that inhibits tumor cell proliferation and angiogenesis and induces cancer cell apoptosis. It also improves the survival rates of patients with advanced liver cancer. However, due to its poor solubility, fast metabolism, and low bioavailability, clinical applications of sorafenib have been substantially restricted. In recent years, various studies have been conducted on the use of nanoparticles to improve drug targeting and therapeutic efficacy in HCC. Moreover, nanoparticles have been extensively explored to improve the therapeutic efficacy of sorafenib, and a variety of nanoparticles, such as polymer, lipid, silica, and metal nanoparticles, have been developed for treating liver cancer. All these new technologies have improved the targeted treatment of HCC by sorafenib and promoted nanomedicines as treatments for HCC. This review provides an overview of hot topics in tumor nanoscience and the latest status of treatments for HCC. It further introduces the current research status of nanoparticle drug delivery systems for treatment of HCC with sorafenib.

Current status of sorafenib nanoparticle delivery systems in the treatment of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is difficult to diagnose early, resulting in only 30% resection rate. HCC is a relatively chemo-resistant tumor, molecular targeted therapy can only benefit approximately 30% patients with liver cancer. Bufalin (Bu) is one of the topoisomerase II inhibitors, many studies have recently focused on the anticancer activities of bufalin. In the present study, we report that bufalin can inhibit the proliferation, invasion and metastasis of liver cancer cells via the Hh signaling pathway. The human high metastasis potential LM3 hepatoma cells (HCC-LM3) were cultured in vitro, bufalin and/or Hedgehog signaling pathway inhibitors (GANT61, cyclopamine) was added into cell culture fluid for 72 h to observe the antitumor effect of bufalin. The results showed that bufalin was able to inhibit epithelial mesenchymal transition (EMT), and extracellular matrix (ECM) degradation and angiogenesis of liver cancer cells by influencing the expression of Ptch1'Gli1'Gli3 proteins in Hh signaling pathway. Bufalin could downregulate the downstream target molecules of MMP-2, MMP-9, β-catenin and VEGF in liver cancer cells by influencing the Gli1 and Gli3 expression of Hh signaling pathway, and upregulate the E-cadherin expression of liver cancer cells by influencing the Gli3 expression of Hh signaling pathway. Therefore, the present study shows that bufalin combined with Hedgehog signaling pathway inhibitors can significantly reduce the malignant biological behavior of the liver cancer cells.

/pdf/inhibitory-effect-of-bufalin-combined-with-hedgehog-4izcci5zyb.pdf

Inhibitory effect of bufalin combined with Hedgehog signaling pathway inhibitors on proliferation and invasion and metastasis of liver cancer cells

Autophagy is an important mechanism which regulates the processes of cell growth and death. The biological role of autophagy in regulating and controlling the proliferation of liver cancer cells by bufalin remains unknown. In the present study we investigated the effect of bufalin on autophagy of liver cancer cells. The growth inhibition and autophagy of liver cancer cells were detected using acridine orange fluorescence staining, flow cytometry and transmission electron microscopy. Combined with autophagy inhibitors (3‑MA and CQ), CCK8 staining and western blot analysis were used to detect the effect of bufalin on the proliferation and autophagy‑related protein expression in HCC‑LM3 cells at the indicated time‑points. Results indicated that combined with autophagy inhibitors 3‑MA and CQ, the inhibitive effect of bufalin on the proliferation of HCC‑LM3 cells was significantly enhanced. When combined with autophagy inhibitors 3‑MA or CQ, bufalin significantly reduced the autophagosome and acidic vesicles in HCC‑LM3 cells. When combined with autophagy inhibitors 3‑MA and/or CQ for 12 h, bufalin significantly inhibited the expression of LC3‑I and Beclin‑1 in HCC‑LM3 cells, and upregulated the expression of LC3‑II and P62 in HCC‑LM3 cells. When combined with autophagy inhibitors 3‑MA and/or CQ for 24 h, bufalin significantly inhibited the LC3‑II expression in HCC‑LM3 cells, and upregulated the LC3‑I, P62 and Beclin‑1 expression in HCC‑LM3 cells. When combined with autophagy inhibitors 3‑MA and/or CQ for 48 h, bufalin significantly inhibited the expression of LC3‑II and Beclin‑1 in HCC‑LM3 cells, and upregulated the expression of LC3‑I and P62 in HCC‑LM3 cells. These findings indicated that bufalin induced cell autophagy and inhibited proliferation of liver cancer cells by influencing the expression of autophagy related proteins including LC3‑I, LC3‑II, P62 and Beclin‑1 in liver cancer cells. The autophagic state of liver cancer cells affected the inhibitory effect of bufalin on the proliferation of liver cancer cells.

https://www.spandidos-publications.com/10.3892/or.2018.6365/download

The biological role of autophagy in regulating and controlling the proliferation of liver cancer cells induced by bufalin.

Sorafenib is a molecularly targeted drug used for treating hepatocellular carcinoma. However, sorafenib may affect the function of normal hepatocytes, and the clinical application of sorafenib is limited due to its adverse effects. The aim of the current study was to improve the effectiveness of sorafenib by preparing it as a nanoparticle formulation using nanoprecipitation technology. Sorafenib was combined with a polyethylene glycol monomethyl ether-racemic polylactic acid copolymer. The properties of the nanoparticles, including particle size, ξ potential and release efficiency, were measured. The pharmacokinetic profile, tissue distribution and tumor-inhibiting effects of the nanoparticles were determined in vitro and in vivo. Compared with sorafenib, the nanoparticle formulation exhibited a significant increase in in vivo retention time. The concentration of sorafenib in tumor tissues was significantly higher than that in normal tissues following treatment with sorafenib nanoparticles. Sorafenib nanoparticles were more efficacious in inhibiting tumor growth compared with sorafenib alone. The results, provided they can be extended to humans, suggest that sorafenib nanoparticles may specifically target hepatocellular carcinoma.

https://www.spandidos-publications.com/10.3892/ol.2017.6934/download

Preparation, pharmacokinetics, tissue distribution and antitumor effect of sorafenib-incorporating nanoparticles in vivo.

In vitro and in vivo studies have demonstrated that brucine is able to inhibit the proliferation of liver cancer cells and growth of animal tumors, and may be a promising anticancer drug. However, high toxicity, poor water solubility, short half-life, narrow therapeutic window, and similar therapeutic and toxic doses limit its clinical application in the treatment of malignant tumors. In our previous study, brucine immuno-nanoparticles were successfully prepared and added to the culture medium of liver cancer SMMC-7721 cells, and the results indicated that the brucine immuno-nanoparticles were able to target the cell membrane of liver cancer SMMC-7721 cells and significantly inhibit the proliferation, adhesion, invasion and metastasis of SMMC-7721 cells. The aim of the present study was to investigate the antitumor effect of brucine immuno-nanoparticles in vivo by establishing an in situ transplanted liver cancer in nude mice. The results indicated that in vivo application of the brucine immuno-nanoparticles resulted in temporary liver and kidney damage, and significantly reduced the α-fetoprotein (AFP) secretion of tumor cells (Bru-NP-MAb vs. the other groups; P<0.05). The brucine concentration of tumor tissues in the brucine immuno-nanoparticles group was significantly increased compared with that of the brucine nanoparticles group (Bru-NP-MAb vs. Bru-NP group or brucine group; P<0.05). The brucine immuno-nanoparticles were able to inhibit tumor growth and cluster of differentiation 34 expression and angiogenesis of tumor tissues, and induce the apoptosis of tumor cells (Bru-NP-MAb vs. Bru-NP group or brucine group; P<0.05). In conclusion, as a novel type of targeted drug, brucine nanoparticles combined with anti-AFP monoclonal antibodies was more effective compared with brucine nanoparticles or brucine alone in inhibiting tumor growth via the enhancement of apoptosis, and the suppression of proliferation and angiogenesis in vivo. Therefore, the brucine immuno-nanoparticle is a promising targeted drug for the treatment of hepatocellular carcinoma.

/pdf/antitumor-effects-of-brucine-immuno-nanoparticles-on-2yhz73ydid.pdf

Antitumor effects of brucine immuno-nanoparticles on hepatocellular carcinoma in vivo

The pathogenesis of hepatocellular carcinoma is complex and not fully known yet. This study aims to screen and identify the differentially expressed proteins in peripheral blood and liver tissue samples from rat hepatocellular carcinoma and to further clarify the pathogenesis and discover the specific tumor markers and molecular targets of hepatocellular carcinoma. The hepatocellular carcinoma model of Wistar rats were induced by chemical carcinogen. The serum and liver tissue samples were obtained after induction for 2, 4, 8, 14, 18, and 21 weeks. The results showed that the clusterin (IPI00198667), heat shock protein a8 (IPI00208205), and N-myc downstream-regulated gene-2 (IPI00382069) being closely related to hepatocarcinogenesis were eventually identified from the 30 different proteins. As the time progressed, the serum levels of clusterin and heat shock protein a8 increased gradually during induced liver cancer in rats. However, the serum N-myc downstream-regulated gene 2 level in induced liver cancer in rats underwent biphasic changes, and the serum N-myc downstream-regulated gene 2 level decreased at the 8th week, increased at the 14th week, and then decreased significantly. Statistical difference occurred in protein expression of clusterin and heat shock protein a8 in liver tissues at the different time points. In the liver tissues, the N-myc downstream-regulated gene 2 level decreased gradually at the 8th week, increased gradually at the 14th week, and then decreased significantly after 14 weeks. The study demonstrated that heat shock protein a8, clusterin, and N-myc downstream-regulated gene 2 participated in the process of abnormal cell division, proliferation, and carcinogenesis of liver cells during hepatocarcinogenesis.

https://journals.sagepub.com/doi/pdf/10.1177/1533034618756785

Xia Sheng

Papers

Inhibitory effect of bufalin combined with Hedgehog signaling pathway inhibitors on proliferation and invasion and metastasis of liver cancer cells

The biological role of autophagy in regulating and controlling the proliferation of liver cancer cells induced by bufalin.

Preparation, pharmacokinetics, tissue distribution and antitumor effect of sorafenib-incorporating nanoparticles in vivo.

Antitumor effects of brucine immuno-nanoparticles on hepatocellular carcinoma in vivo

Identification of the Differential Expression Profiles of Serum and Tissue Proteins During Rat Hepatocarcinogenesis.