Effects of annexin A5 knockdown on cell cycle of human gastric cancer MGC-803 and MKN-45 cells

doi:10.16098/j.issn.0529-1356.2018.03.012

Abstract

Abstract:

Objective To investigate the effects of annexin A5 (ANXA5) konckdown on cell cycle of human gastric cancer cell lines MGC-803, MKN-45 cells. Methods Cells were diveded into interference group, negative control group and blank control group. The siRNA targeted to ANXA5 was transfected into MGC-803, MKN-45 cells by lipofectamine 2000 respectively. At the same time, negative siRNA was also transfected into negative control group and in blank control group nothing was added in. Real-time PCR and Western blotting was used to identify whether the siRNA targeted to ANXA5 can highly suppress annexin A5 expression 48 hours after transfection. When the effect of the siRNA was assured, Real-time PCR and Western blotting were applied to observe the expression of p21^cip1 mRNA and P21^cip1 protein, Western blotting was used to detect the expression of cyclinD1, flow cytometry(FCM) was applied to observe the cell cycle progression. Results The expression of p21^cip1 mRNA, P21^cip1 protein and cyclinD1 decreased significantly when compared with the negative control group and the blank control group(P<0.05). FCM analysis showed that the percentage of cells in G1 phase increased in interference group(P<0.05). Conclusion ANXA5 knockdown can down-regulate the expression of p21^cip1 mRNA, P21^cip1 protein and cyclinD1 in human gastric cancer MGC-803 and MKN-45 cells, it is speculated that ANXA5 may affect the development of gastric cancer by inducing cell cycle arrest by acting on cell cycle related proteins.

Key words: Annexin A5, MGC-803, MKN-45, CyclinD1, Real-time PCR, Western blotting, Flow cytometry, Human

WANG Xiao-jie ZHAO Yi-na LI Xin. Effects of annexin A5 knockdown on cell cycle of human gastric cancer MGC-803 and MKN-45 cells [J]. Acta Anatomica Sinica, 2018, 49(3): 342-346.

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