miR-578调控长链脂酰辅酶A合成酶4表达对胃癌细胞增殖、侵袭的影响★

Effects of miR-578 regulating the expression of long-chain acyl-CoA synthetase 4 on the proliferation and invasion of gastric cancer cells★

摘要:

目的 探讨miR-578调控长链脂酰辅酶A合成酶4（ACSL4）对胃癌细胞增殖及侵袭的影响。方法 选取人胃癌细胞MKN-45、SNU-1、MKN-7、KTAO3、N87及人正常胃黏膜细胞GES-1为研究对象，将MKN-45细胞分为对照组、miR-578 NC组、miR-578 mimic组、miR-578 mimic+pc-ACSL4组，测定各组MKN-45细胞活力及凋亡、侵袭、迁移能力，测定细胞中miR-578、ACSL4、凋亡相关蛋白（Bax、Caspase-3、Bcl-2）、侵袭相关蛋白（MMP-9、MMP-2）的表达，验证miR-578与ACSL4的靶向关系。结果 miR-578在人胃癌细胞中低表达；与对照组相比，miR-578 mimic组MKN-45细胞活力、侵袭、迁移能力、MMP-2、MMP-9、Bcl-2蛋白表达水平均显著降低（P<0.05），凋亡能力、Caspase-3、Bax蛋白表达水平均显著升高（P<0.05）；与miR-578 mimic组相比，miR-578 mimic+pc-ACSL4组MKN-45细胞活力、侵袭、迁移能力、MMP-2、MMP-9、Bcl-2蛋白表达水平均显著升高（P<0.05），凋亡能力、Caspase-3、Bax蛋白表达水平均显著降低（P<0.05）；荧光素酶实验结果显示，miR-578与ACSL4存在靶向调节关系。结论 miR-578可抑制人胃癌细胞MKN-45的增殖及迁移能力，可能是通过抑制ACSL4表达实现的。

Abstract:

Objective To investigate the effects of miR-578 regulating long-chain acyl-CoA synthetase 4 (ACSL4) on the proliferation and invasion of gastric cancer cells.Methods Human gastric cancer cells MKN-45, SNU-1, MKN-7, KTAO3, N87, and human normal gastric mucosal cells GES-1 were selected as the objects of the study. The MKN-45 cells were divided into control group, miR-578 NC group, miR-578 mimic group, miR-578 mimic + pc-ACSL4 group. Detect the viability, apoptosis ability, invasion ability, migration ability of MKN-45 cells in each group, and determine the expressions of miR-578, ACSL4, apoptosis-related proteins (Bax, Caspase-3, Bcl-2) and invasion-related proteins (MMP-9, MMP-2) in cells, and verify the targeting relationship between miR-578 and ACSL4.Results miR-578 showed low expression in human gastric cancer cells. Compared with the control group, the viability, invasion and migration abilities of MKN-45 cells in the miR-578 mimic group was attenuated significantly, and the expression levels of MMP-2, MMP-9 and Bcl-2 proteins were decreased significantly (P<0.05), while the apoptosis ability and the expression levels of Caspase-3 and Bax proteins were significantly increased (P<0.05). Compared with the miR-578 mimic group, the MKN-45 cell viability, invasion and migration abilities of MKN-45 cells in the miR-578 mimic+pc-ACSL4 group was enhanced significantly, and the expression levels of MMP-2, MMP-9 and Bcl-2 proteins were increased (P<0.05), while the apoptosis ability and the expression levels of Caspase-3 and Bax proteins were decreased (P<0.05). The results of luciferase assay showed that miR-578 and ACSL4 had a targeted regulation relationship.Conclusion miR-578 can inhibit the proliferation and migration of human gastric cancer cell MKN-45 by suppressing the expression of ACSL4.