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Archive > Volume 24 Issue 11 > 2017,24(11):1260-1265. DOI:10.3872/j.issn.1007-385X.2017.11.003 Prev Next
Basic Research
Effect of silencing Gli2 gene on proliferation and apoptosis of the hepatocellular carcinoma SMMC-7721 line cell
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Abstract:
Objective: To explore effect of silencing glioma associated oncogene homolog 2 (Gli2) on proliferation and apoptosis of the liver cancer SMMC-7721 cell and its possible action mechanism. Methods: Infection of the SMMC-7721 cell with recombinant shRNA-Gli2 lentivirus was used to screen the best jamming sequence for silence effect. The SMMC-7721 cell of interference group was infected with the best jamming sequence lentivirus,the SMMC-7721 cell of control group was infected with shRNA-NC shRNA-Gli2 lentivirus and the SMMC-7721 cell of blank group didn’t with any treatment. CCK-8 and Western blotting assays were used to detect proliferation activity of the SMMC-7721 cell in various groups. Flow cytometry assay was used to test apoptosis of the SMMC-7721 cell in various groups. Expressions of Cli2, cyclinD1, Bax and Bcl-2 mRNA and their proteins were detected by qPCR and Western blotting. Results: Infection rate of the recombinant lentivirus was about 90%. Silence effect of the recombinant shRNA-Gli2-1 lentivirus was the best. Expressions of Gli2 mRNA and its protein in the SMMC-7721 cell of interference group were remarkably lower than those in the SMMC-7721 cells of control and blank groups (all P<0.05), and differences of Gli2 mRNA and its protein expressions between the SMMC-7721 cells of control group and blank group were not significant (P>0.05). Proliferation and cell colony forming number of the SSMC-7721 cell in interference group were obviously less than those of the SSMC-7721 cell in control and blank groups (all P<0.05). Apoptosis rate of the SMMC-7721 cell in interference group evidently higher than those of the SMMC-7721 cell in control and blank groups (P<0.01). Expressions of cyclinD1 and Bax mRNA and their proteins in the SMMC-7721 cell of interference group were observably lower than those in the SMMC-7721 cells of control and blank groups (all P<0.05), but expressions of Bcl-2 mRNA and protein in the SMMC-7721 cell of interference group were observably higher than those in the SMMC-7721 cells of control and blank groups (all P<0.05). Conclusion:Silencing expression of Gli2 gene could inhibit growth of the liver cancer SMMC-7721 cell, and promote apoptosis of the liver cancer cell, its mechanism might relate to down-regulation of cyclin D1 and Bax as well as upregulation of Bcl-2.
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Project Supported:
Project supported by the Scientific Research Foundation of the Health and Family Planning Commission of Sichuan Province (No.17PJ590)
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