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Effect of GP73 silencing on proliferation and apoptosis in hepatocellular cancer
Yu-Long Zhang1, You-Cheng Zhang1, Wei Han1
1Yu-Long Zhang, You-Cheng Zhang, Wei Han, Yu-Min Li, Geng-Nian Wang, Shao Yuan, Feng-Xian Wei, Jia-Feng Wang, Jian-Jun Jiang, Ya-Wu Zhang, Department of General Surgery, Lanzhou University Second Hospital, Lanzhou 730030, Gansu Province, China.
Aim:
To investigate the roles of Golgi protein (GP) 73 in the regulation of cell proliferation and apoptosis.
Methods:
Stealth RNAi targeting GP73 gene sequence was used to silence its expression in Hep G2 cells and Bel7402 cells. Stealth RNAi effects were assessed by reverse transcriptase polymerase chain reaction and ELISA. Cell proliferation assay and cell cycle analysis were assessed by MTT assay and flow cytometry. Apoptosis was assessed by flow cytometry and transmission electron microscopy. Apoptosis-related proteins were assessed by western immunoblot analysis.
Results:
Stealth RNAi targeting GP73 gene sequence markedly reduced the expression of GP73 gene. The reduction of GP73 in Hep G2 cells and Bel7402 cells inhibited cell proliferation and induced apoptosis, however, terminal apoptosis occurred in Hep G2 cells, but early apoptosis occurred in Bel7402 cells. Reduced expression of GP73 gene might lead to a reduction in Bcl-2/Bax ratio, an increase in cytochrome c, but a reduction in capase-3.
Conclusion:
GP73 might play an important role in proliferation and apoptosis in hepatocellular carcinoma cells.
Insights
Golgi protein 73 (GP73) silencing inhibited hepatocellular carcinoma cell proliferation and induced apoptosis. GP73 plays a key role in regulating cell growth and programmed cell death in liver cancer.
Area of Science:
- Molecular Biology
- Cell Biology
- Oncology
Background:
- Golgi protein 73 (GP73) is implicated in various cellular processes.
- Understanding GP73's role in hepatocellular carcinoma (HCC) is crucial for therapeutic development.
Purpose of the Study:
- To investigate the function of GP73 in regulating cell proliferation and apoptosis in HCC cells.
- To elucidate the molecular mechanisms underlying GP73's effects on cell fate.
Main Methods:
- Gene silencing of GP73 using Stealth RNAi in Hep G2 and Bel7402 HCC cell lines.
- Assessed gene expression, cell proliferation (MTT assay), cell cycle (flow cytometry), and apoptosis (flow cytometry, transmission electron microscopy).
- Analyzed apoptosis-related proteins (Western immunoblot analysis).
Main Results:
- GP73 gene silencing significantly reduced GP73 expression, inhibiting cell proliferation and inducing apoptosis in both cell lines.
- Hep G2 cells exhibited terminal apoptosis, while Bel7402 cells showed early apoptosis.
- Reduced GP73 expression altered the Bcl-2/Bax ratio, increased cytochrome c release, and decreased caspase-3 activity.
Conclusions:
- GP73 plays a significant role in the proliferation and apoptosis of hepatocellular carcinoma cells.
- Modulating GP73 expression presents a potential therapeutic strategy for HCC.
