膀胱癌T24细胞顺铂与吉西他滨多药耐药细胞株的建立及其生物学特征  

Establishment of multidrug resistance cell line of cisplatin and gemcitabine in bladder cancer cellline T24 and its biological characteristics

作　　者：王鹏 陈明坤 陈子坚 郭晓彬 杨诚 卞军 周其赵 郭文彬 薛康颐 张万松 杨建昆 刘存东 Wang Peng;Chen Mingkun;Chen Zifian;Guo Xiaobin;Yang Cheng;Bian Jun;Zhou Qizhao;Guo Wenbing;Xue Kangyi;Zhang Wansong;Yang Jiankun;Liu Cundong(Department of Urology, the Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China)

机构地区：[1]南方医科大学第三附属医院泌尿外科,广州510630

出　　处：《中华实验外科杂志》2018年第4期716-718,共3页Chinese Journal of Experimental Surgery

基　　金：广东省自然科学基金（2014A030310017）

摘　　要：目的采用浓度梯度法诱导建立顺铂（DDP）及吉西他滨（GEM）多药耐药膀胱癌细胞株（T24/DDP＆GEM），并分析其生物学特征。 方法利用人膀胱癌细胞株T24，采用DDP和GEM作为诱导剂在体外建立DDP及GEM多药耐药的人膀胱癌细胞模型T24/DDP＆GEM。通过倒置相差显微镜下观察T24、T24/DDP＆GEM细胞形态差异，细胞计数试剂盒（CCK-8）方法检测相关细胞的耐药性，生长曲线检测相关膀胱癌细胞的增殖能力，Western blot检测P-糖蛋白（P-gP）的表达。 结果经过DDP和GEM 12个月的交替的诱导，成功建立了T24/DDP＆GEM。倒置相差显微镜下观察并与T24细胞比较，T24DDP/GEM形态不规则，体积变大，细胞内可见空泡和颗粒形成。T24/DDP＆GEM的增殖能力与T24比较显著下降，T24/DDP＆GEM的倍增时间为（39.51±0.36） h，T24的倍增时间为（28.06±0.76） h，倍增时间延长（11.45±0.41） h。T24/DDP＆GEM的耐药性明显提高，其耐药指数对于DDP为4.25，对于GEM为62.97。与T24细胞比较，T24/DDP＆GEM细胞的S和G2期的细胞比例显著减少，而G1期细胞比例显著增多，细胞周期减慢。Western blot结果显示，T24/DDP＆GEM细胞中P-gp的表达水平明显高于T24。 结论人工诱导的DDP及GEM耐药膀胱癌细胞株T24/DDP＆GEM具有多药耐药肿瘤细胞特征。ObjectiveTo establish a human bladder cancer cisplatin （DDP） and gemcitabine （GEM） resistant cell line （T24/DDP＆GEM） by concentration gradient method, and to observe its biological characteristics. MethodsDDP and GEM were used as inducers to establish human bladder cancer cisplatin and gemcitabine resistant model T24/DDP＆GEM in vitro. The morphology of T24 and T24/DDP＆GEM cells was observed by the inverted microscope. The drug resistance and the growth curve of cells were detected by cell counting kit-8 （CCK-8）. The P-glycoprotein （P-gP） expression was detected by Western blotting. ResultsAfter the induction of DDP and GEM for 12 months, T24/DDP＆GEM was successfully established. Under the inverted phase contrast microscopy, as compared with T24 cells, the morphology of T24DDP/GEM cells was irregular, the volume became larger, and vacuoles and granules were formed in the cytoplasm. The proliferation ability of T24/DDP＆GEM cells was significantly lower than that of T24 cells. The doubling time of T24/DDP＆GEM cells was （39.51±0.36） h, and that of T24 was （28.06±0.76） h. The doubling time was extended for （11.45±0.41） h. The drug resistance of T24/DDP＆GEM was significantly improved, and the drug resistance index was 4.25 for DDP and 62.97 for GEM. As compared with T24 cells, the proportion of S and G2 phase cells in T24/DDP＆GEM cells decreased, the proportion of G1 cells increased significantly, and the cell cycle slowed down. The results of Western blotting showed that the expression level of P-gp in T24/DDP＆GEM cells was significantly higher than that in T24 cells. ConclusionThe artificial induced DDP and GEM resistant bladder cancer cell line T24/DDP＆GEM has the characteristics of multi drug resistant tumor cells.

关 键 词：膀胱癌 化疗 顺铂 吉西他滨 多药耐药 

分 类 号：R737.14[医药卫生—肿瘤]