雷公藤红素对人结直肠癌细胞HCT-8增殖和侵袭迁移的影响及作用机制  

Effects of celastrol on proliferation,invasion and migration of human colorectal cancer HCT-8 cells and its mechanism

作　　者：张宇涵 李香 张玲 马燕春 ZHANG Yuhan;LI Xiang;ZHANG Ling;MA Yanchun(Translational Medicine Research Center,Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西医科大学转化医学研究中心,太原030001

出　　处：《山西医科大学学报》2023年第9期1176-1182,共7页Journal of Shanxi Medical University

基　　金：中国博士后科学基金资助项目(2019M651071);国家自然科学基金资助项目(81802440);山西省研究生教育创新项目(2022Y400)。

摘　　要：目的探究雷公藤红素(celastrol,Cel)对结直肠癌细胞株HCT-8增殖和转移能力的影响及相关分子机制。方法分别以不同浓度的Cel(0,0.5,1.0,2.0μmol/L)处理HCT-8细胞24 h。采用MTT法检测细胞的存活率;细胞集落克隆形成法检测细胞增殖;Transwell法检测细胞侵袭迁移能力;活性氧检测试剂盒检测细胞内ROS水平;Western blot检测细胞凋亡相关蛋白(cleaved PARP、Bcl-2、Bax)、丝裂原活化蛋白激酶(MAPK)信号通路及上皮间质转化(EMT)相关蛋白的相对表达量。结果MTT结果显示,细胞的存活率随Cel浓度增加而下降(P<0.01);Transwell结果显示,细胞侵袭和迁移能力随Cel浓度增加而下降(P<0.001);细胞内ROS含量随Cel浓度增加而增多,并且Western blot结果显示Bax、cleaved PARP、p-JNK、p-p38 MAPK、E-cadherin蛋白表达随Cel浓度增加而升高(P<0.01),Bcl-2、p-ERK1/2、Vimentin、Twist、Slug、Snail蛋白表达均显著降低并呈浓度依赖性(P<0.01)。结论雷公藤红素可以抑制结直肠癌HCT-8细胞的增殖、侵袭和转移,其作用机制可能与MAPK信号通路引起细胞凋亡及上皮间质转化进程(EMT)有关。Objective To explore the effects of celastrol(Cel)on the proliferation and metastasis of HCT-8 cells and the related molecular mechanism.Methods HCT-8 cells were treated with Cel at different concentrations(0,0.5,1.0,2.0μmol/L)for 24 h.The survival rate of HCT-8 cells treated with celastrol was detected by MTT assay,and its effect on the cell proliferation was detected by colony cloning assays.Transwell method was used to detect the cell invasion and migration.The reactive oxygen species detection kit was used to detect the level of intracellular ROS.Western blot was used to measure the relative expression of apoptosis-related proteins(cleaved PARP,Bcl-2,Bax),the mitogen-activated protein kinase(MAPK)signaling pathway-related and EMT-associated marker proteins in HCT-8 cells.Results MTT and Transwell results showed that the cell survival rate and the cell invasion and migration abilities were statistically decreased by Cel in a concentration-dependent manner(P<0.01).The intracellular ROS content and and the relative expression levels of Bax,cleaved PARP,p-JNK/p-p38 MAPK and E-cadherin were increased by Cel in a concentration-dependent manner(P<0.01),and the relative expression levels of Bcl-2,p-ERK1/2,Vimentin,Twist,Slug and Snail were decreased significantly in a concentration-dependent manner(P<0.01).Conclusion Celastrol can inhibit the proliferation,the invasion and the metastasis of HCT-8 cells,which is related to the apoptosis and EMT through MAPK signaling pathway.

关 键 词：雷公藤红素 HCT-8细胞 MAPK信号通路 细胞增殖 细胞凋亡 

分 类 号：R735.3[医药卫生—肿瘤]