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Effects of silencing B7-H4 expression by small interference RNA on proliferation, invasion and migration of human lung adenocarcinoma A549 cells
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Abstract:
Objective:To investigate the effects of silencing B7-H4 expression by small interference RNA (siRNA) on proliferation, invasion and migration of human lung adenocarcinoma A549 cells. Methods: Chemically synthesized siRNA targeting B7-H4 (B7-H4-siRNA) was transfected into A549 cells. The proliferation of A549 cells was determined by MTT assay, the cell cycle was detected by flow cytometry, the levels of B7-H4 and Cyclin D1 were verified by Western blotting, and the invasion and migration ability was detected by Transwell assay. Results: B7-H4-siRNA was successfully transfected into A549 cells. Western blotting results showed that B7-H4-siRNA transfection inhibited the expressions of B7-H4 and Cyclin D1 in A549 cells. In the B7-H4-siRNA group, the proliferation of A549 cells was significantly down-regulated, the doubling time was longer than that in the untransfected group, the Ctrl-siRNA group and the empty vector group (\[33.78±0.26\] h vs \[28.69±0.18\], \[27.32±0.13\], \[26.93±0.19\] h，P<0.05), and the cell cycles were arrested in G1 phase. The invasion of A549 cells in the B7-H4-siRNA group was inhibited significantly as compared with the untransfected group (\[89.80±0.99\] vs \[186.20±1.33\], P<0.05), and the migration of A549 cells in the B7-H4-siRNA group was suppressed significantly as compared with the untransfected group, the Ctrl-siRNA group and the empty vector group (\[60.20±0.37\] vs \[102.57±0.52\], \[100.72±0.31\], \[98.65±0.21\], P<0.05). Conclusion: B7-H4-siRNA can silence B7-H4 expression in A549 cells, and inhibit the proliferation, invasion and migration of A549 cells effectively. B7-H4 can be regarded as a candidate gene for lung cancer gene therapy.
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