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Lentivirusmediated shRNA silencing of livin gene promotes apoptosis of SPC-A1 cells
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Abstract:
Objective:To construct a lentiviral livin shRNA vector to silence livin gene expression, and to study its effect on apoptosis of lung carcinoma cells. Methods:Livin expression in human lung adenocarcina SPCA1 cells was silenced by lentiviral livin shRNA infection. The morphology of apoptotic cells was observed by propidine iodide staining and fluoroscope; apoptosis rate and sublipliod apoptotic peak of SPCA1 cells were assessed by flow cytometry; expression of livin and caspase 3 in SPCA1 cells was examined by realtime PCR and Western blotting analysis. Results: Livin was constitutively expressed in SPCA1 cells. After livin expression was silenced by lentiviral livin shRNA infection, SPCA1 cells showed the characteristic morphology of apoptosis under fluoroscope, and the sublipliod apoptotic peak was identified by flow cytometry. Apoptosis rate in livin shRNA infected SPCA1 cells was significantly higher than that in blank and negative control groups (8.3% vs 0.08% and 0.13%, P<0.05). caspase 3 mRNA expression in SPCA1 cells had no change but the expression of cleavedcaspase 3 was greatly upregulated after lentiviral livin shRNA infection as showed by RTPCR and Western blotting analysis. Conclusion: Lentiviral livin shRNA can inhibit livin expression in human lung adenocarcina SPCA1 cells and induce cell apopotosis.
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Project Supported:
Supported by the Natural Science Foundation of Fujian Province（No. 2008J0074 ）
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