RAD5对结肠癌HT29细胞的抗肿瘤活性及相关机制初步探索  

Exploration of the antitumor activity and related mechanisms of RAD5 on colon cancer HT29 Cells

作　　者：韦立群 徐成飞 潘晓杭 阮国添 闫岭 唐双意[1] 甘嘉亮[2] Wei Liqun;Xu Chengfei;Pan Xiaohang;Ruan Guotian;Yan Ling;Tang Shuangyi;Gan Jialiang(Department of Pharmacy,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China;Colorectal&Anal Surgery,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学第一附属医院药学部,广西南宁530021 [2]广西医科大学第一附属医院结直肠肛门外科,广西南宁530021

出　　处：《结直肠肛门外科》2024年第3期322-328,共7页Journal of Colorectal & Anal Surgery

基　　金：广西壮族自治区卫生健康委员会自筹经费科研课题(Z20180959)。

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(0μmol/L、12.5μmol/L、25μmol/L、50μmol/L、100μmol/L、150μmol/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(0μmol/L)、25μmol/L、50μmol/L的RAD5干预36 h,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0μmol/L相比,25μmol/L组、50μmol/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。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(0μmol/L,12.5μmol/L,25μmol/L,50μmol/L,100μmol/L,150μmol/L)for 36 hours,and cell viability was detected using the MTT assay.Colon cancer HT29 cells were divided into 3 groups and treated with RAD5 for 36 hours with 0μmol/L,25μmol/L and 50μmol/L respectively,the cell proliferation ability was assessed using a colony formation assay,apoptosis rate was detected by Hoechst 33258 staining and flow cytometry,and the expression of apoptosisrelated proteins Bax,Bcl-2,and EGFR/MAPK pathway-related proteins was examined by Western blot.Results The results 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(0μmol/L),the clone formation rates of the 25μmol/L and 50μmol/L groups were significantly reduced(P<0.001),apoptosis rates increased(P<0.01),Bax expression increased,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 expression and activating the phosphorylation of ERK1/2 and P38 in the downstream MAPK pathway,providing experimental theoretical support for its development as an anticancer candidate drug.

关 键 词：结肠癌 迷迭香酸衍生物 增殖 凋亡 机制 

分 类 号：R735.3[医药卫生—肿瘤]