SH-6联合CHKA基因沉默对胶质瘤细胞增殖及侵袭的影响

Effects of SH-6 combined with CHKA gene silencing on proliferation and invasion of glioma cells

目的:探讨特异性AKT抑制剂SH-6联合慢病毒沉默CHKA基因在体内外对胶质瘤细胞系U87增殖、侵袭的影响及机制。方法:构建CHKA短发夹CHKA(shCHKA)慢病毒及对照慢病毒(shNC)感染U87细胞系并设置分组。分别采用CCK-8法检测胶质瘤细胞的增殖,流式细胞术检测胶质瘤细胞的周期、凋亡,Transwell实验观察胶质瘤细胞侵袭。胶质瘤动物模型实验评估联合治疗效果并记录肿瘤体积变化,采用免疫组化及Western blot检测胶质瘤细胞及肿瘤组织中CHKA、PI3K/AKT信号通路中相关蛋白的表达。结果:联合治疗能显著抑制胶质瘤细胞的增殖及侵袭,细胞周期主要停滞在G2期,细胞凋亡率显著增加,联合治疗能够使CHKA、PI3K/AKT信号通路中相关蛋白的表达量降低。结论:SH-6联合慢病毒沉默CHKA在体内外能够显著抑制U87细胞的增殖及侵袭。本研究可能为脑胶质瘤的联合治疗提供新的抗肿瘤策略。

Objective:To investigate the effect and mechanism of specific AKT inhibitor SH-6 combined with lentiviral silencing of CHKA gene on proliferation and invasion of human glioma cell line U87 in vivo and in vitro.Methods:CHKA short hairpin CHKA(shCHKA)lentivirus and control lentivirus(SHNC)were constructed to infect U87 cell line and set up for grouping. hairpin CHKA(shCHKA)lentivirus and control lentivirus(SHNC)were constructed to infect U87 cell line and set up for grouping. The proliferation of glioma cells was detected by CCK-8 method,the cell cycle and apoptosis of glioma cells were detected by flow cytometry,and the biological functions of glioma cells were observed by Transwell assay. Glioma animal model experiments were used to evaluate the effect of combined treatment and record the changes in tumor volume. Immunohistochemistry and Western blot were used to detect the expressions of CHKA and PI3K/AKT signaling pathway-related proteins in glioma cells and tumor tissues.Results:The combination treatment significantly inhibited glioma cell proliferation and invasion,cell cycle was mainly arrested in the G2phase,and the apoptosis rate was significantly increased. The combination treatment resulted in a reduction in the expressions of CHKA,related proteins in the PI3K/AKT signalling pathway.Conclusion:Silencing of CHKA by SH-6 combined with lentivirus significantly inhibites U87 cell proliferation and invasion in vitro and in vivo. The present study may provide a new antitumor strategy for the combination therapy of brain gliomas.