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Archive > Volume 20 Issue 6 > 2013,20(6):706-710. DOI:10.3872/j.issn.1007-385X.2013.06.013 Prev Next
Basic Research
Reduction of cisplatin resistance of lung cancer A549 cells through down-regulating the expression of BAG-1 mediated by shRNA
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Abstract:
Objective : To detect down regulation of the expression of BAG-1 gene in lung cancer A549 by transfected a vector contained shRNA, and to investigate its effects on the cisplatin (DDP) resistance of A549 cells. Methods: Interference vector pGCsi-BAG-1 target BAG-1 was constructed and stable transferred into A549 cells. Cells stable transferred by pGCsi-BAG-1 were used as an experimental group (BAG-1-shRNA), cells transferred by non-sense vector were used as a negative control group (SC-shRNA), and the parent A549 cells were used as a control group. Western blotting was performed to detect the effect of pGCsi-BAG-1 transfection on the BCL-2 and BAG-1 expression of A549 cells. MTT method and flow cytometry was used to detect the influence on proliferation and apoptosis of A549 cells after DDP treatment. Results: An A549 cell line where BAG-1 was stably interfered was successfully constructed. And the expression of BCL-2 protein in cells of the BAG-1- shRNA group was significantly lower than that of SC-shRNA group and the control group (all P<005). With increasing of the concentration of DDP, the proliferation inhibition rate of each cell was increased. When DDP concentration was 2.5 μg/ml, the cell proliferation inhibition rate of the BAG-1-shRNA group was significantly higher than that of SC-shRNA group and the control group (\[8.12±4.09\] % vs \[10.07± 3.82\] %, \[22.26±489\]%; all P<0.05). Compared with the SC-shRNA group and the control group, the apoptosis rate of the BAG-1-shRNA group was significantly increased after 24 h treatment with DDP (\[37.84±3.62\]% vs \[16.80±2.81\]%, \[1710±311\]%, P< 0.05). Conclusion: Down-regulation of BAG-1 expression can inhibit the proliferation of A549 cells after DDP treatment and promote its apoptosis.
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Project Supported:
Project supported by the Science and Technology Plan Projects of Liaoning Province （No. 2007225005-13）
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