Sorafenib-induced Aquaporin-3 Downregulation is Coupled With Proliferation Inhibition and Increased Apoptosis in Hepatocellular Carcinoma Cells

2020 ◽  
Author(s):  
Kija Malale ◽  
Jili Fu ◽  
Liewang Qiu ◽  
Ke Zhan ◽  
Xiuni Gan ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Cell Proliferation ◽  
Cell Cycle Arrest ◽  
Western Blotting ◽  
Proliferation Inhibition ◽  
Cyclin Dependent Kinase ◽  
Aquaporin 3 ◽  
Cycle Arrest ◽  
Hcc Cells

Abstract Background Sorafenib is the only targeted therapy promising to improve the prognosis of patients with advanced hepatocellular carcinoma (HCC), but its long-term clinical efficacy is limited due to chemotherapy resistance. The lack of a full understanding of the anti-tumor mechanism of sorafenib in HCC is attributed to the difficulties in understanding the mechanism of drug resistance. In recent years, a large number of preclinical and clinical data have confirmed the catalytic role of aquaporin-3 (AQP3) in a variety of tumors including HCC, but none of the studies reported the regulatory mechanism of AQP3 during sorafenib treatment. This study examined the effect of sorafenib on the expression of AQP3 in HCC cells and determined whether the effect is associated with cell proliferation inhibition, cell cycle arrest, and increased apoptotic. Methods mRNA and protein levels of AQP3 in hepatoma cell lines exposed to sorafenib or UO126 were detected via real-time quantitative polymerase chain reaction (qPCR) and western blotting, respectively. The effect of AQP3 expression changes on cell proliferation, cell cycle and apoptosis were determined by cell counting kit-8 (CCK-8) and flow cytometry. In addition, western blotting detected proteins involved in the regulation of proliferation and cell cycle progression. Results The results showed that AQP3 was down-regulated in all cell lines exposed to sorafenib or UO126 in a concentration dependent manner. The downregulation of AQP3 successfully inhibited cell proliferation, induced cell cycle arrest and increased cell apoptosis, while the reverse was true when AQP3 was overexpressed. Western blotting results showed that in AQP3 knockdown cells, the amounts of Erk, Akt, p53, p-Erk, p-Akt and cyclin-dependent kinase 2 (CDK2) decreased, while the amounts of cyclin-dependent kinase 4 (CDK4), p21 and p-p53 increased. Conclusion This study found that sorafenib may inhibit proliferation, induce cell cycle arrest, and increase apoptosis of HCC cells by regulating the expression of AQP3.

scholarly journals Knockout of Hepatocyte Growth Factor by CRISPR/Cas9 System Induces Apoptosis in Hepatocellular Carcinoma Cells

2021 ◽  
Vol 11 (10) ◽  
pp. 983
Author(s):  
Han Ki Lee ◽  
Heui Min Lim ◽  
See-Hyoung Park ◽  
Myeong Jin Nam
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Cell Proliferation ◽  
Growth Factor ◽  
Hepatocyte Growth Factor ◽  
Cell Cycle Arrest ◽  
Cycle Arrest ◽  
Apoptotic Effects ◽  
Hepatocyte Growth ◽  
Hcc Cells

Background: CRISPR/Cas9 system is a prokaryotic adaptive immune response system that uses noncoding RNAs to guide the Cas9 nuclease to induce site-specific DNA cleavage. Hepatocyte growth factor (HGF) is a well-known growth factor that plays a crucial role in cell growth and organ development. According to recent studies, it has been reported that HGF promoted growth of hepatocellular carcinoma (HCC) cells. Here, we investigated the apoptotic effects in HCC cells. Methods: Crispr-HGF plasmid was constructed using GeneArt CRISPR Nuclease Vector. pMex-HGF plasmid that targets HGF overexpressing gene were designed with pMex-neo plasmid. We performed real time-polymerase chain reaction to measure the expression of HGF mRNA. We performed cell counting assay and colony formation assay to evaluate cell proliferation. We also carried out migration assay and invasion assay to reveal the inhibitory effects of Crispr-HGF in HCC cells. Furthermore, we performed cell cycle analysis to detect transfection of Crispr-HGF induced cell cycle arrest. Collectively, we performed annexin V/PI staining assay and Western blot assay. Results: In Crispr-HGF-transfected group, the mRNA expression levels of HGF were markedly downregulated compared to pMex-HGF-transfected group. Moreover, Crispr-HGF inhibited cell viability in HCC cells. We detected that wound area and invaded cells were suppressed in Crispr-HGF-transfected cells. The results showed that transfection of Crispr-HGF induced cell cycle arrest and apoptosis in HCC cells. Expression of the phosphorylation of mitogen activated protein kinases and c-Met protein was regulated in Crispr-HGF-transfected group. Interestingly, we found that the expression of HGF protein in conditioned media significantly decreased in Crispr-HGF-transfected group. Conclusions: Taken together, we found that inhibition of HGF through transfection of Crispr-HGF suppressed cell proliferation and induced apoptotic effects in HCC Huh7 and Hep3B cells.

scholarly journals Novel Ferrocene Derivatives Induce G0/G1 Cell Cycle Arrest and Apoptosis through the Mitochondrial Pathway in Human Hepatocellular Carcinoma

2021 ◽  
Vol 22 (6) ◽  
pp. 3097
Author(s):  
Jianrong Zheng ◽  
Liao Zeng ◽  
Mingqing Tang ◽  
Hongjun Lin ◽  
Chao Pi ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Cell Cycle Arrest ◽  
Mitochondrial Pathway ◽  
Proliferation Inhibition ◽  
Ferrocene Derivatives ◽  
Cycle Arrest ◽  
Human Liver Cell ◽  
G1 Cell Cycle Arrest ◽  
Hcc Cells

In this study, detailed information on hepatocellular carcinoma (HCC) cells (HepG-2, SMMC-7721, and HuH-7) and normal human liver cell L02 treated by ferrocene derivatives (compounds 1, 2 and 3) is provided. The cell viability assay showed that compound 1 presented the most potent and selective anti-HCC activity. Further mechanism study indicated that the proliferation inhibition effect of compound 1 was associated with the cycle arrest at the G0/G1 phase and downregulation of cyclin D1/CDK4. Moreover, compound 1 could induce apoptosis in HCC cells by loss of mitochondrial membrane potential (ΔΨm), accumulation of reactive oxygen species (ROS), decrease in Bcl-2, increase in BAX and Bad, translocation of Cytochrome c, activation of Caspase-9, -3, and cleavage of PARP. These results indicated that compound 1 would be a promising candidate against HCC through G0/G1 cell cycle arrest-related proliferation inhibition and mitochondrial pathway-dependent apoptosis.

scholarly journals The Novel Nature Microtubule Inhibitor Ivalin Induces G2/M Arrest and Apoptosis in Human Hepatocellular Carcinoma SMMC-7721 Cells In Vitro

Medicina ◽  
2019 ◽  
Vol 55 (8) ◽  
pp. 470 ◽  
Author(s):  
Fangyuan Liu ◽  
Shiqi Lin ◽  
Caiyun Zhang ◽  
Jiahui Ma ◽  
Zhuo Han ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Cell Cycle Arrest ◽  
Western Blotting ◽  
Network Formation ◽  
Cyclin B1 ◽  
Human Hepatocellular Carcinoma ◽  
Microtubule Depolymerization ◽  
Microtubule Network ◽  
Cycle Arrest

Background and Objectives: Microtubules are an attractive target for cancer chemotherapy. Previously, we reported that Ivalin exhibited excellent anti-migration and anti-invasion activities in human breast cancer cells. Here, we examined the microtubule inhibition effect of Ivalin in human hepatocellular carcinoma SMMC-7721 cells. Materials and Methods: We used the 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay to evaluate the cell proliferation effect of Ivalin and flow cytometry analysis to detect the apoptotic and cell cycle arrest effects of Ivalin. Immunofluorescence staining was used to measure the effect of Ivalin on the cytoskeleton network, and Western blotting was used to detect the expression levels of Bax, Bcl-2, Cdc2, phosphor-Cdc2, Cdc25A, Cyclin B1, and tubulin. Results: Ivalin induced cell cycle G2/M arrest and subsequent triggered apoptosis in human hepatocellular carcinoma SMMC-7721 cells. Furthermore, microtubules were shown to be involved in Ivalin-meditated apoptosis. In this connection, Ivalin treatment suppressed cellular microtubule network formation by regulating microtubule depolymerization. Moreover, Western blotting revealed Cdc25A and Cyclin B1 were upregulated in Ivalin-meditated cell cycle arrest. Subsequently, the induction of Bax (a proapoptotic protein) and reduction of Bcl-2 (an anti-apoptotic protein) expression were observed in Ivalin-treated SMMC-7721 cells. Conclusion: Ivalin induced microtubule depolymerization, then blocked cells in mitotic phase, and eventually resulted in apoptosis in SMMC-7721 cells. Collectively, these data indicate that Ivalin, acting as a novel inhibitor of microtubules, could be considered as a promising lead in anticancer drug development.

scholarly journals Melatonin enhances the anti-tumor effect of sorafenib via AKT/p27-mediated cell cycle arrest in hepatocarcinoma cell lines

RSC Advances ◽  
2017 ◽  
Vol 7 (34) ◽  
pp. 21342-21351 ◽  
Author(s):  
Fei Long ◽  
Chengyong Dong ◽  
Keqiu Jiang ◽  
Yakun Xu ◽  
Xinming Chi ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Cell Cycle Arrest ◽  
Cell Lines ◽  
Cycle Arrest ◽  
Hepatocarcinoma Cell ◽  
Proposed Model ◽  
Hcc Cells

Proposed model elucidating the role of MT in regulating the proliferation of hepatocellular carcinoma (HCC) cells treated with sorafenib.

scholarly journals Dysregulation of miR-23b-5p promotes cell proliferation via targeting FOXM1 in hepatocellular carcinoma

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Xinchen Yang ◽  
Shikun Yang ◽  
Jinhua Song ◽  
Wenjie Yang ◽  
Yang Ji ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Cell Proliferation ◽  
Cell Cycle Arrest ◽  
Tumor Development ◽  
Loss Of Function ◽  
Cycle Arrest ◽  
Cell Proliferation Inhibition

AbstractGrowing evidence demonstrates that MicroRNAs (miRNAs) play an essential role in contributing to tumor development and progression. However, the underlying role and mechanisms of miR-23b-5p in hepatocellular carcinoma (HCC) formation remain unclear. Our study showed that miR-23b-5p was downregulated in the HCC tissues and cell lines, and lower expression of miR-23b-5p was associated with more severe tumor size and poorer survival. Gain- or loss-of-function assays demonstrated that miR-23b-5p induced G0/G1 cell cycle arrest and inhibited cell proliferation both in vitro and in vivo. qRT-PCR, western blot and luciferase assays verified that Mammalian transcription factor Forkhead Box M1 (FOXM1), upregulated in HCC specimens, was negatively correlated with miR-23b-5p expression and acted as a direct downstream target of miR-23b-5p. In addition, miR-23b-5p could regulate cyclin D1 and c-MYC expression by directly targeting FOXM1. Further study revealed that restoration of FOXM1 neutralized the cell cycle arrest and cell proliferation inhibition caused by miR-23b-5p. Taken together, our findings suggest that miR-23b-5p acted as a tumor suppressor role in HCC progression by targeting FOXM1 and may serve as a potential novel biomarker for HCC diagnosis and prognosis.

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