Akt抑制剂MK-2206在缺氧环境下对结肠癌SW480细胞增殖、侵袭的影响

The effects of Akt inhibitor MK-2206 on proliferation and invasion of colon cancer SW480 cells under hypoxia condition

目的:探讨Akt抑制剂MK-2206在缺氧环境下对人结肠癌SW480细胞增殖、侵袭的影响。方法:根据CCK-8实验结果选择CoCl_(2)和MK-2206的浓度,最后分成空白组、MK-2206组、CoCl_(2)组、MK-2206+CoCl_(2)组;Transwell小室实验检测四组细胞的迁移和侵袭能力;RT-PCR法检测各细胞组中Akt、mTOR、HIF-1α的mRNA表达水平;Western blot技术检测各细胞组中Akt、p-Akt、mTOR、p-mTOR、HIF-1α蛋白表达的差异性情况。结果:CoCl_(2)诱导的缺氧环境可以促进SW480细胞的侵袭、迁移(P<0.05),同时能够促进HIF-1α的mRNA和蛋白表达(P<0.05),但是低浓度范围内的CoCl_(2)对SW480细胞增殖活性无明显影响(P>0.05);MK-2206可以在体外的常氧和缺氧环境中抑制SW480细胞的增殖活性和侵袭、迁移能力(P<0.05);MK-2206能在体外的常氧环境下抑制SW480细胞的Akt、mTOR、HIF-1α的mRNA表达(P<0.05),缺氧环境无明显作用;同时MK-2206能够在常氧和缺氧环境中显著降低p-Akt、p-mTOR、HIF-1α的蛋白表达水平(P<0.05)。结论:Akt抑制剂MK-2206可以在体外的常氧和缺氧环境下抑制SW480细胞的增殖活性和侵袭能力,但缺氧环境可能会弱化MK-2206对SW480细胞迁移的抑制作用。

Objective:To investigate the effects of Akt inhibitor MK-2206 on the proliferation and invasion of human colon cancer SW480 cells under hypoxia condition.Methods:According to the results of CCK-8 experiment and the concentrations of CoCl_(2) and MK-2206,there were blank group,MK-2206 group,CoCl_(2) group and MK-2206+CoCl_(2) group.Transwell chamber test was used to detect the ability of cell migration and invasion,and RT-PCR technique was used to detect the mRNA expression of Akt,mTOR and HIF-1αin each cell group.Western blot was used to detect the difference of Akt,p-Akt,mTOR,p-mTOR and HIF-1αprotein expression in different cell groups.Results:CoCl_(2)-induced anoxic environment could not only promote the invasion and migration of SW480 cells(P<0.05),but also promote the expression of mRNA and protein of HIF-1α(P<0.05).However,CoCl_(2) at low concentration had no significant effect on the proliferation of SW480 cells(P>0.05).MK-2206 can inhibit the proliferation,invasion and migration of SW480 cells in normoxic and hypoxic environment(P<0.05).MK-2206 could inhibit the mRNA expression of Akt,mTOR and HIF-1αin SW480 cells under normoxic environment in vitro(P<0.05),but had no effect in hypoxic environment.At the same time,MK-2206 could significantly reduce the protein expression levels of p-Akt,p-mTOR and HIF-1αin normoxic and hypoxic environment(P<0.05).Conclusion:MK-2206 of Akt inhibitor can significantly inhibit the proliferation and invasion of SW480 cells under normoxic and hypoxic conditions in vitro.However,hypoxia may weaken the inhibitory effect of MK-2206 on the migration of SW480 cells.