膀胱癌T24顺铂耐药细胞系的建立及生物学特性  

Establishment of Cisplatin-multidrug Resistance Cell Line of Human Bladder Cancer T24 and Its Biological Characteristics

作　　者：赵嫚[1] 张曼[1,2] 马萍[1] 

机构地区：[1]徐州医学院医学技术学院,江苏徐州221004 [2]首都医科大学附属北京世纪坛医院,北京100038

出　　处：《标记免疫分析与临床》2015年第4期338-341,共4页Labeled Immunoassays and Clinical Medicine

基　　金：北京市中医管理局中西医结合项目(2014-ZYJ04)

摘　　要：目的采用浓度梯度法诱导建立人膀胱癌顺铂耐药细胞系(T24/DDP),并观察其生物学特性。方法利用人膀胱癌细胞株T24,采用顺铂(DDP)作为诱导剂在体外建立人膀胱癌顺铂耐药细胞模型T24/DDP(0.6μg/m L)。通过倒置相差显微镜下观察T24、T24/DDP细胞形态,MTT法检测细胞多药耐药性,生长曲线检测细胞的增殖能力,流式细胞术测细胞的周期,q RT-PCR检测耐药基因MRP(multidrug resistance-associated protein)的表达。结果经过顺铂14个月的诱导,成功建立了T24/DDP。倒置相差显微镜下观察与T24细胞相比较,T24/DDP排列不规则,形态出现多形性,出现巨细胞;与T24相比较,T24/DDP对顺铂、阿霉素、长春新碱、甲氨蝶呤均产生不同程度的交叉耐药,比T24细胞倍增时间明显延长,细胞周期中G1期细胞增多,G2、S期细胞减少。q RT-PCR结果显示,耐药细胞MRP基因的表达明显高于亲本细胞。结论人膀胱癌耐顺铂细胞株T24/DDP具有多药耐药性,可以作为研究膀胱癌细胞多药耐药的良好实验模型。Objective To establish a cisplatin resistant bladder cancer cell line and to analysis its biological characteristics. Methods Drug concentration gradient increasing method was adopted to induce drug resistance. T24 and T24/DDP were observed under phase contrast microscope. Multidrug resistance Of the two cell lines was determined by MT-F. Cell proliferation was evaluated according to the growth curve. Flow cytometry was performed to determine cycles of the two cell lines. The expression of multidrug resistance related gene muhidrug resistance-associated protein（MRP） was detected by qRT-PCR. Results After 14 months, cisplatin- resistant human bladder cancer cell line T24/DDP was successfully established from its parental cell line T24. Two kinds of ceils were significantly different from each other by a microscope. T24/DDP cell was 9.51 times more resistant to DDP than T24. T24/DDP cell line exhibited the characteristics of cross-resistance to cisplatin, adriamycin, methotrexate, vincristine. The cell doubling time of T24/DDP was longer than parental cell line T24. Compared with T24, the proportion of G2 and S phase cells in T24/DDP decreased significantly, while that of G1 phase cells increased significantly. The expression of drug-resistance related gene MRP was significantly up-regulated in T24/DDP （P 〈 0. 05 ）. Conclusion With the characteristics of muhidrug resistance, cisplatin-resistant human bladder cancer cell line T24/DDP could be used as a good experimental model for further study on bladder cancer multidrug resistance.

关 键 词：人膀胱癌细胞 顺铂 多药耐药 

分 类 号：R737.14[医药卫生—肿瘤]