摘要
目的探究细胞周期蛋白依赖性激酶7(Cyclin-Dependent Kinase 7,CDK7)抑制剂THZ1(CDK7共价抑制剂)对人结直肠癌SW480细胞增殖、迁移及凋亡的影响。方法通过体外培养人结直肠癌细胞系SW480细胞,CCK8法测定细胞增殖活力以筛选THZ1对SW480细胞的作用浓度。将SW480细胞分为对照组(加入相应体积的DMSO)、低、中、高浓度组(处理后THZ1药物浓度为20、80、160 nmol/L)。平板克隆实验法检测细胞克隆能力测定THZ1对SW480细胞克隆形成的影响;划痕实验分别测定THZ1对SW480细胞迁移能力的影响,TUNEL法检测细胞凋亡情况;蛋白质印迹法检测CDK-7、p-CDK7、Bax、Bcl-2、Mcl-1、Parp蛋白表达水平。结果SW480细胞增殖抑制率在药物浓度大于80 nmol/L时显著升高(P<0.05),低、中、高浓度组SW480细胞克隆能力低于对照组,差异有统计学意义(P<0.05),低浓度组的划痕愈合率低于对照组,中浓度组的划痕愈合率低于对照组,高浓度组的划痕愈合率低于对照组,差异有统计学意义(P<0.05),低、中、高浓度组的凋亡率高于对照组,差异有统计学意义(P<0.05),高浓度组CDK-7、p-CDK7、Bcl-2、Mcl-1蛋白表达水平低于对照组,高浓度组Bax、Parp蛋白表达水平高于对照组,差异均有统计学意义(P<0.05)。说明THZ1可促进SW480细胞凋亡,抑制其增殖和迁移。结论CDK7抑制剂THZ1可以抑制人结直肠癌SW480细胞增殖和迁移能力,并诱导细胞凋亡。
Objective To explore the effects of CDK7 inhibitor THZ1 on the proliferation,migration and apoptosis of human colorectal cancer SW480 cells.Methods Human colorectal cancer cell line SW480 was cultured in vitro,and the cell proliferation activity was determined by CCK8 method to screen the effect concentration of THZ1 on SW480 cells.SW480 cells were divided into control group(adding corresponding volume of DMSO),low concentration and high concentration groups with the concentration of THZ120,80,160 nmol/L,respectively.The effect of THZ1 on colony formation of SW480 cells was determined by experimental method of plate cloning.The effect of THZ1 on the migration ability of SW480 cells was determined by test of scratches.Cell apoptosis was detected by TUNEL method.The protein expression levels of CDK-7,p-CDK7,Bax,Bcl-2,Mcl-1 and Parp were detected by western blot method.Results The inhibition rate of SW480 cell proliferation was significantly increased when the drug concentration was greater than 80 nmol/L(P<0.05).The cloning abilities of SW480 cells in low and high concentration groups were significantly lower than those in control group(P<0.05).The scratch healing rate of the low concentration group was lower than that of the control group.The scratch healing rate of the medium concentration group was lower than that of the control group,and the scratch healing rate of the high concentration group was lower than that of the control group,the difference was statistically significant(P<0.05).The apoptosis rates of the low and high concentration groups were higher than those of the control group,and the difference was statistically significant(P<0.05).The expression levels of CDK-7,p-CDK7,Bcl-2 and Mcl-1 protein in the high concentration group were lower than those in the control group,and the expression levels of Bax and Parp protein in the high concentration group were higher than those in the control group,with statistical significance(P<0.05).These results indicated that THZ1 could promote apoptosis and inhibit pr
作者
王建哲
赵奇
于伟光
贾信奇
赵岩松
李敏捷
范东
姜浩
WANG Jian-zhe(Graduate School of Mudanjiang Medical College,Mudanjiang 157011,China)
出处
《牡丹江医学院学报》
2023年第2期29-33,4,共6页
Journal of Mudanjiang Medical University
基金
牡丹江市级科技指导性计划项目(HT2022NS082)。
