摘要
目的 探究磷脂酰肌醇3激酶(PI3K)/哺乳动物雷帕霉素靶蛋白(mTOR)抑制剂BEZ235联合顺铂对膀胱癌细胞迁移和侵袭的影响,及其潜在的作用机制。方法 在2020年1月1日至2021年5月30日,应用细胞计数试剂盒(Cell Counting Kit-8,CCK-8)法检测BEZ235和顺铂联合接单药对人膀胱癌细胞(5637细胞)增殖的影响,并计算联合用药指数(CI);在BEZ235和顺铂空白处理(对照组)、联合处理和单药分别处理膀胱癌5637细胞后,通过平板克隆形成实验、划痕愈合实验和细胞侵袭实验分别检测各处理组膀胱癌细胞的集落形成、迁移和侵袭能力;应用蛋白质印迹法检测各处理组的膀胱癌细胞中上皮-间质转化相关蛋白的表达。结果 BEZ235和顺铂对5637细胞均以时间和剂量依赖的方式发挥抑制作用,100 nmol/L BEZ235和0.1μmol/L顺铂联合用药时,CI值最小。BEZ235和顺铂联合处理组的5637细胞的集落形成数[对照组为(207±9)个,BEZ235单药处理组为(146±10)个,顺铂单药处理组为(179±11)个,两药联合处理组为(103±9)个]、相对迁移率[对照组为(70±5)%,BEZ235单药处理组为(52±6)%,顺铂单药处理组为(53±4)%,两药联合处理组为(39±6)%]和侵袭数[对照组为(346±36)个,BEZ235单药处理组为(193±38)个,顺铂单药处理组为(254±27)个,两药联合处理组为(133±23)个]均显著少于对照组和各单药组(均P<0.05)。BEZ235和顺铂联合处理组的5637细胞中上皮-间质转化相关蛋白上皮钙黏素(E-cadherin)的表达水平均显著高于对照组和各单药组(均P<0.05),神经钙黏素(N-cadherin)和波形蛋白的表达水平均显著低于对照组和各单药组(均P<0.05)。结论 BEZ235和顺铂可在体外协同抑制膀胱癌细胞的增殖,并通过上皮-间质转化途径抑制细胞的迁移和侵袭。
Objective To explore the effect of Phosphatidylinositol 3 kinase(PI3K)/mammalian target of rapamycin(mTOR) inhibitor BEZ235 combined with cisplatin on migration and invasion of bladder cancer cells and its potential mechanism. Methods From January 1, 2020 to May 30, 2021, the effects of BEZ235 and cisplatin on the proliferation of bladder cancer 5637 cells were detected by CCK-8, and the combination index(CI) was calculated. After blank treatment(control group), combined treatment and single treatment with BEZ235 and cisplatin, the colony formation, migration and invasion ability of bladder cancer cells in each treatment group were detected by plate clone formation assay, scratch healing assay and cell invasion assay, respectively. Western blotting was used to detect the expression of epithelial-mesenchymal transition-related proteins in bladder cancer cells of each treatment group.Results BEZ235 and cisplatin inhibited 5637 cells in a time-dependent and dose-dependent manner. The CI value was the lowest when 100 nmol/L BEZ235 and 0.1 μmol/L cisplatin were combinedly used. Colony formation number [(207±9) in the control group,(146±10) in the BEZ235 monotherapy group,(179±11) in the cisplatin monotherapy group, and(103±9) in the combined treatment group], relative mobility [(70±5)% in the control group,(52±6)% in the BEZ235 monotherapy group,(53±4)% in the cisplatin monotherapy group, and(39±6)% in the combined treatment group] and the number of invasions [(346±36) in the control group,(193±38) in the BEZ235 monotherapy group,(254±27) in the cisplatin monotherapy group, and(133±23) in the combined treatment group] of 5637 cells in the combined treatment group were significantly lower than those in control group and respective monotherapy groups(all P<0.05). The expression of epithelial-mesenchymal transition-related protein E-cadherin in 5637 cells treated with BEZ235 and cisplatin was significantly higher than that in the control group and each monotherapy group(all P<0.05), while the expression leve
作者
陈康
邢基
张云龙
程帆
CHEN Kang;XING Ji;ZHANG Yunlong;CHENG Fan(Department of Urinary Surgery,Renmin Hospital of Wuhan University,Wuhan,Hubei 430060,China)
出处
《安徽医药》
CAS
2023年第3期443-447,I0001,共6页
Anhui Medical and Pharmaceutical Journal
基金
湖北省科技重大专项项目(2019AEA170)。
关键词
膀胱肿瘤
细胞增殖
BEZ235
顺铂
上皮-间质转化
Urinary bladder neoplasms
Cell proliferationl
BEZ235
Cisplatin
Epithelial-mesenchymal transformation
