摘要
目的 探究RAD5对结肠癌HT29细胞的抗肿瘤活性及其可能机制.方法 用不同浓度RAD5(0 μmol/L、12.5μmol/L、25 μmol/L、50 μmol/L、100 µmol/L、150 μmol/L)干预结肠癌HT29细胞36 h,采用MTT法检测细胞存活率.将结肠癌HT29细胞分为3组,分别给予(0 μmol/L)、25 μmol/L、50 μmol/L的RAD5干预36h,采用克隆形成实验检测细胞增殖能力,Hoechst 33258染色和流式细胞术检测细胞凋亡率,Western blot检测细胞凋亡蛋白Bax、Bcl-2以及EGFR/MAPK通路相关蛋白表达.结果细胞实验结果表明,RAD5能显著抑制结肠癌HT29细胞增殖且有浓度依赖性.与对照组0 μmol/L相比,25 μmol/L组、50 μmol/L组的结肠癌HT29细胞克隆形成率明显降低(P＜0.001),细胞凋亡率增加(P＜0.01),Bax表达增加,Bcl-2表达有下降趋势,EGFR/MAPK 通路中 EGFR、ERK1/2 表达明显降低,p-ERK1/2/ERK1/2 和 p-P38/P38 升高(P＜0.001).结论 RAD5 对结肠癌HT29细胞具有抗肿瘤活性,其机制可能与抑制EGFR表达、激活下游MAPK通路中ERK1/2、P38磷酸化有关,为开发其作为抗肿瘤候选药物提供了实验理论参考.
Abstract
Objectives To investigate the antitumor activity and potential mechanisms of rosmarinic acid derivative 5(RAD5)on colon cancer HT29 cells.Methods Colon cancer HT29 cells were treated with different concentrations of RAD5(0 μmol/L,12.5 μmol/L,25 μmol/L,50 μmol/L,100 μmol/L,150 μmol/L)for 36 hours,and cell viability was de-tected using the MTT assay.Colon cancer HT29 cells were divided into 3 groups and treated with RAD5 for 36 hours with 0 μmol/L,25 μmol/L and 50 μmol/L respectively,the cell proliferation ability was assessed using a colony forma-tion assay,apoptosis rate was detected by Hoechst 33258 staining and flow cytometry,and the expression of apoptosis-related proteins Bax,Bcl-2,and EGFR/MAPK pathway-related proteins was examined by Western blot.Results The re-sults of cell experiments showed that RAD5 could significantly inhibit the proliferation of colon cancer HT29 cells in a concentration-dependent manner.Compared with the control group(0 μmol/L),the clone formation rates of the 25 μmol/L and 50 μmol/L groups were significantly reduced(P＜0.001),apoptosis rates increased(P＜0.01),Bax expression in-creased,Bcl-2 expression showed a downward trend,and the expression of EGFR and ERK1/2 proteins in the EGFR/MAPK pathway decreased,while the ratios of p-ERK1/2/ERK1/2 and p-P38/P38 were elevated(P＜0.001).Conclusion RAD5 exhibits antitumor activity against colon cancer HT29 cell,and its mechanism may be related to inhibiting EGFR ex-pression and activating the phosphorylation of ERK1/2 and P38 in the downstream MAPK pathway,providing experimen-tal theoretical support for its development as an anticancer candidate drug.
基金项目
广西壮族自治区卫生健康委员会自筹经费科研课题(Z20180959)
维普
万方数据