Mobile website
Basic Research
Inhibitory effects of BRCAA1 gene silencing on gastric cancer MGC803 cells and its possible mechanism
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective:To investigate the inhibitory effect of breast cancerassociated antigen 1(BRCAA1) gene silencing on gastric cancer MGC803 cells and the related mechanism. Methods: Plasmid shRNABRCAA1 and shRNAN were constructed and transfected with FuGene HD into gastric cancer cell line MGC803. The transfection efficiency was examined using fluorescent microscope 24 h later. The total RNAs was extracted 48 h after transfection and the expression of BRCAA1 and GAPDH gene were analyzed by realtime PCR. The cell proliferation was assessed by MTT assay 24 h, 48 h, and 72 h after transfection. The cell apoptosis was determined by Annexin VPE/7AAD. The expression of Rb, Bax, Bcl2 and BRCAA1 proteins was analyzed by Western blotting 48 h after transfection. Results:We found that the transfection efficiency of shRNABRCAA1 was （81.2±2.6）％ 24 h after transfection. Fortyeight hours after transfection with shRNABRCAA1 the expression of BRCAA1 mRNA decreased by 61.4%; the inhibition rate of MGC803 cells growth was 45.0%. The cell apoptosis rate of shRNABRCAA1 transfection group was significantly higher than those of untransfected group and mock plasmid transfected group （\[14.4±1.6\]% vs \[5.4±2.0\]％，\[4.4±2.5\]％，P<0.05\]. Cells transfected with shRNABRCAA1 had significantly increased expression of Rb and Bax proteins（P<0.05）, and decreased expression of Bcl2 protein（P<005）. Conclusion: BRCAA1 gene silencing can effectively inhibit the proliferation of MGC803 cells and induce apoptosis, which might be related to the promotion of Rb and Bax proteins, and suppression of Bcl2 protein.
Keywords:
Project Supported:
Supported by the Major State Basic Research Development Program (973) of China (No.2005CB723400G); the National High Technology Research and Development Program (863) of China (No.2007AA022004); the National Natural Science Foundation of China (No.307710
Clc Number:
