GSTP1基因对人肝癌细胞系HepG2增殖和侵袭能力的影响

刘润田; 安聪静; 白云; 郑建兴

doi:10.3971/j.issn.1000-8578.2016.12.006

GSTP1基因对人肝癌细胞系HepG2增殖和侵袭能力的影响

Effects of GSTP1 on Proliferation and Invasion of Human HepG2 Cells

摘要

摘要:
目的探讨过表达/沉默GSTP1基因对人肝癌细胞系HepG2增殖及侵袭能力的影响。
方法采用腺病毒载体转染人肝癌细胞系HepG2，获得过表达/沉默GSTP1基因的HepG2细胞。实时荧光定量PCR（qPCR）法检测细胞中GSTP1 mRNA表达水平；CCK-8法和Transwell小室法分别检测细胞的细胞活力和侵袭能力；免疫印迹法（Western blot）检测细胞中Akt、mTOR、p-Akt和p-mTOR的蛋白表达。
结果经腺病毒载体转染后，成功获得过表达/沉默GSTP1基因的HepG2细胞；过表达GSTP1基因后，HepG2细胞的细胞活力和侵袭能力显著降低（P<0.05），而沉默GSTP1基因后，HepG2细胞的细胞活力和侵袭能力则显著增高（P<0.05）；过表达GSTP1基因后，p-Akt和p-mTOR的蛋白表达水平显著降低（P<0.05），而沉默GSTP1基因的结果则相反。
结论过表达GSTP1基因可抑制HepG2细胞的增殖和侵袭能力，沉默GSTP1基因则促进HepG2细胞的增殖和侵袭能力，其作用机制可能与Akt/mTOR信号通路有关。

Abstract:
Objective To investigate the effect of GSPT1 overexpression or knockdown on the proliferation and invasion of human HepG2 cells.
Methods HepG2 cells were infected with adenovirus (Ad)-GSTP- or Ad-shGSTP1 to overexpress or knockdown the expression of GSTP1 in HepG2 cells. The expression level of GSTP1 mRNA was examined by quantitative PCR; CCK-8 and Transwell assay were used to determine the cell viability and invasion, respectively. The expression of Akt, mTOR, p-Akt and p-mTOR were detected by Western blot.
Results The mRNA expression of GSTP1 was up-regulated after infection with Ad-GSTP1, and was down-regulated after infection with Ad-shGSTP1(P<0.05). The overexpression of GSTP1 significantly inhibited the viability and invasion of HepG2 cells(P<0.05), however, the knockdown of GSTP1 enhanced cell viability and invasion(P<0.05). Moreover, the phosphorylation level of Akt and mTOR were increased in the cells infected with Ad-GSTP1(P<0.05). Inverse results were obtained in the cells infected with Ad-shGSTP1.
Conclusion The viability and invasion of HepG2 cells could be inhibited by GSTP1 overexpression while enhanced by GSTP1 knockdown, which may be related with Akt/mTOR pathway.

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