下调人四旋蛋白1表达后奥沙利铂耐药结肠癌细胞雌激素受体相关受体α水平变化  

Expression and change of ERRα inoxaliplatin-resistant colon cancer cells after the downregulation of tetrasoanin

作　　者：朱佳 闫晓菲[2] 李晓曦[3] 张睿[2] 石刚[2] 刘信[2] 陈悦[2] 廉波[2] 马思平[2] 刘放[2] 

机构地区：[1]中国医科大学肿瘤医院辽宁省肿瘤医院内镜中心,沈阳110042 [2]中国医科大学肿瘤医院结直肠外科,沈阳110042 [3]中国医科大学肿瘤医院中心实验室,沈阳110042

出　　处：《中国综合临床》2018年第1期9-14,共6页Clinical Medicine of China

基　　金：国家自然科学基金资助项目（81672427）;辽宁省博士科研启动基金（201601417）

摘　　要：目的观察下调人四旋蛋白1表达后奥沙利铂耐药结肠癌细胞株增殖及凋亡及雌激素受体相关受体（estrogen receptor related receptor，ERR）α水平变化。方法以Western blot法和实时定量（qRT）-PCR法检测结肠癌细胞及奥沙利铂耐药细胞中人四旋蛋白1及ERRα在蛋白和mRNA水平表达，以ERRα抑制剂XCT790下调ERRα表达，Western blot法、流式细胞术及MTT检测L-OHP-SW620细胞人四旋蛋白1、凋亡及增殖。采用siRNA下调人四旋蛋白1表达，Western blot法、qRT-PCR、流式细胞术及MTT检测L-OHP-SW620细胞AKT，p-AKT，人四旋蛋白1及ERRα表达、凋亡及增殖。结果人四旋蛋白1及ERRα蛋白在L-OHP-SW620细胞株中表达高于SW620细胞（t＝6.127，P〈0.01，t＝12.579，P〈0.01），TSPAN1mRNA在L-OHP-SW620细胞株中表达高于SW620细胞（t＝9.085，P〈0.01）。XCT790抑制ERRα表达后XCT790组L-OHP-SW620细胞早期凋亡率高于阴性对照（NC）组（t＝3.297，P〈0.01）；XCT790组细胞培养72 h后生存率为（45.264±6.249）%，低于阴性对照组（63.364±9.472）% （t＝4.537，P〈0.01）。siTSPAN1组与NC组对比，p-AKT蛋白水平上调（t＝8.139，P〈0.01），ERRα蛋白水平下调（t＝6.452，P〈0.01），小干扰人四旋蛋白1组细胞早期凋亡率为（17.541±2.317）%，低于NC组（32.892±3.296）% （t＝4.526，P〈0.01），小干扰人四旋蛋白1组在培养72 h后细胞生存率为（49.653±5.945）%，低于NC组（67.376±7.834）% （t＝3.109，P〈0.05）。结论下调人四旋蛋白1表达上调p-AKT蛋白，降低ERRα蛋白表达，奥沙利铂耐药细胞株增殖降低，凋亡增加，耐药性减弱。ObjectiveTo observe the proliferation and apoptosis of oxaliplatin-resistant colon cancer cell lines and the expression of estrogen receptor related receptor （ERR）α when tetrasoanin was down-regulated.MethodsWestern blot and quantitative real-time polymerase chain reaction（qRT-PCR） were used to detect the expression of tetrasoanin and ERRα of colon cancer cells and oxaliplatin resistant cells in mRNA and protein levels. ERRα inhibitor XCT790 was used to down-regulate ERRα expression, The expression of ERRα was down regulated by ERR α inhibitor XCT790, and the level of tetrasoanin, apoptosis and proliferation of L-OHP-SW620 cells were detected by Western blot, flow cytometry and MTT.Tetrasoanin expression was down regulated by siRNA, the expression, apoptosis and proliferation of L-OHP-SW620 cells AKT, p-AKT, tetrasoanin and ERRα were detected by Western blot, qRT-PCR, flow cytometry and MTT assay.ResultsThe expression of tetrasoanin and ERRα protein in L-OHP-SW620 cell lines were higher than those in SW620 cells （t=6.127, P〈0.01, t=12.579, P〈0.01）, The expression of tetrasoanin mRNA in L-OHP-SW620 cell line was higher than that in SW620 cell line （t=9.085, P〈 0.01）. The early apoptosis rate of L-OHP-SW620 cells in XCT790 group after XCT790 inhibited ERR -αexpression was higher than that in NC group （t=3.297, P〈 0.01）. The survival rate of XCT790 group after 72 h culture was （45.264±6.249）%, lower than that of NC group （ （63.364±9.472）%） （t=4.537, P〈0.01）. Compared with NC group, p-AKT protein was up-regulated（t=8.139, P〈0.01）, ERRα protein was down-regulated（t=6.452, P〈0.01）, the apoptosis rate was（17.541±2.317）%, lower than that in the sitetrasoanin group （（32.892±3.296）%） （t=4.526, P〈0.01）, the survival in sitetrasoanin group after 72 h culture was （49.653±5.945）%, lower than that in NC group（（67.376±7.934）%） （t=3.109, P〈0.05）.ConclusionTetrasoanin down-regulation and p-AKT protein up-regulation decreas

关 键 词：人四旋蛋白1 化疗耐药 结肠癌 小干扰RNA 

分 类 号：R735.35[医药卫生—肿瘤]