新型 CDK7抑制剂对人胶质瘤细胞生物学功能的影响

摘要

目的：探究新型 CDK7抑制剂 THZ1对人胶质瘤细胞系 U87及 U251增殖、凋亡、侵袭能力的影响及细胞周期的阻滞作用。方法：将 U87及 U251细胞分成对照组及实验组，实验组加入不同浓度的 THZ1处理后，以 CCK －8法及平板克隆形成试验分析细胞增殖；Transwell 侵袭试验观测各组侵袭能力的变化；流式细胞仪检测各组细胞周期分布及凋亡情况；Western blot 实验检测凋亡相关蛋白 Caspase －3的表达量。结果：THZ1抑制 U87及 U251的增殖，根据实验结果用 SPSS 20．0分析出72小时药物 IC50，U87细胞为83．1nmol／L、U251细胞为13．7nmol／L。平板克隆形成试验结果显示低浓度药物即可明显抑制克隆形成。THZ1降低U87细胞的侵袭性。THZ1促进 U251细胞凋亡，Caspase －3表达增加。THZ1阻滞 U87及 U251细胞周期进展，G2期细胞显著增加。结论：THZ1能够抑制胶质瘤细胞 U87及 U251的增殖，促进凋亡，抑制侵袭，并阻滞细胞周期进展。%Objective:To study the effects of a new CDK7 inhibitor THZ1 on the proliferation,apoptosis,invasion and cycle of human glioma U87 and U251 cells in vitro.Methods:The U87 and U251 cells were divided into blank group,control group and experimental group.The blank group was added with DMEM complete culture medium.The control group was added with DMEMcomplete culture medium containing 1 /1 000 DMSO (sulfoxide dimethyl).The experimental group was treated with different concentrations of THZ1.The inhibitory effect of THZ1 on cell prolifera-tion was examined by CCK -8(cell counting kit 8)and colony formation assay.The invasion ability was evaluated by Transwell assay.The cell cycle distribution and apoptosis rate of U87 and U251 cells induced by THZ1 were detected by flow cytometry(FCM).The expression of apoptosis related protein Caspase -3 was detected by Western blot assay. Results:THZ1 inhibits the proliferation of U87 and U251:After treatment in each group for 72 hours,there was no significant difference between the blank group and the control group (P >0.05).Each concentration of experimental group was able to inhibit the proliferation of cells,and with the increase of THZ1 concentration,the inhibition degree is more obvious.According to the experimental results,SPSS 20.0 analysis was used to analyze the drug IC50 (half maximal inhibitory concentration),and IC50 of U87 cells were 83.1nmol/L and U251 cells were 13.7nmol/L.The plate clone formation test results show that even very low concentration of drug could inhibit colony formation rate. THZ1 decreases the invasiveness of U87 cells.THZ1 promotes apoptosis of U251 cells.After THZ1 treatment for 12 hours,apoptosis was detected by flow cytometry.The apoptosis rate of the experimental group was significantly in-creased.The expression of apoptosis related protein Caspase -3 was increased.THZ1 block U87 and U251 cell cycle progression:the proportion of G2 phase cells increased significantly,compared with the control group,the difference was statistically significant (P <0.05).Conclusion:THZ1 can inhibit the proliferation,promote apoptosis,inhibit the ability of invasion and block cell cycle progression of U87 and U251 in glioma cells.