哌立福新抑制肝癌HePG2细胞增殖的机制研究

The mechanism of inhibits the proliferation of hepatocellular carcinoma HepG2 cells

目的探讨哌立福新对肝癌HepG2细胞增殖、侵袭和迁移的影响及其机制。方法以不同浓度(0、7.5、15和30μmol/L)的哌立福新处理肝癌HepG2细胞后,MTT法和集落形成实验检测细胞的增殖能力,Transwell小室实验检测细胞侵袭能力,细胞划痕愈合实验检测细胞的迁移能力,葡萄糖和ATP检测试剂盒检测HepG2细胞内葡萄糖和ATP含量变化,Western blot检测增殖细胞核抗原(PCNA)、基质金属蛋白酶9(MMP⁃9)、葡萄糖转运蛋白1(GLUT1)、Akt和mTOR蛋白的表达。结果哌立福新能够呈浓度-时间依赖性抑制肝癌HepG2细胞增殖。随着哌立福新浓度的增加,HepG2细胞的侵袭和迁移能力逐渐减弱,细胞内葡萄糖和ATP含量逐渐减少,PCNA、MMP⁃9、GLUT1、Akt和mTOR蛋白的表达逐渐降低,与对照组(0μmol/L)相比,差异均有统计学意义(P<0.05)。结论哌立福新能够抑制肝癌HepG2细胞的增殖,其作用机制可能与抑制Akt/mTOR途径减弱糖代谢过程有关。

Objective To investigate the effects of perifosine on proliferation,invasion and migration of HepG2 cells and its mechanism.Methods After treating the liver cancer HepG2 cells with different concentrations(0,7.5,15 and 30μmol/L)of perifosine,the proliferation ability of the cells was detected by MTT method and colony formation test.The invasive ability of the cells was detected by the transwell chamber test.The cell migration ability was detected by the scar healing test.The changes of intracellular glucose and ATP content were detected by the glucose and ATP detection kit,and the expression of proliferating cell nuclear antigen(PCNA),matrix metalloprotein 9(MMP⁃9),gulcose transporter protein 1(GLUT1),Akt and mTOR proteins were detected by Western blot.Results The proliferation of HepG2 cells could be inhibited by perifosine in a concentration time dependent manner.With the increase of the concentration of perifosine,the invasion and migration of HepG2 cells were weakened,and the contents of glucose and ATP in the cells were decreased,and the expressions of PCNA,MMP⁃9,GLUT1,Akt and mTOR were decreased gradually,compared with the control group(0μmol/L),the differences were statistically significant(P<0.05).Conclusion Perifosine could inhibit the proliferation of HepG2 cells,which might be related to the reduction of glucose metabolism by the inhibition of Akt/mTOR pathway.