体内诱导法建立的Tca8113细胞耐卡铂细胞株的耐药性表达研究

Multidrug Resistance of an in vivo Induced Carboplatin-Resistant Tca8113 Cell Line

目的探讨通过体内诱导法得到的耐药细胞株的耐药性表达情况。方法以人舌癌Tca8113细胞系为研究对象,实验组采用荷瘤耐药动物的肿瘤细胞培养获得,对照组为体外连续培养诱导其产生耐药性。用四唑蓝法检测细胞耐药指数,标记生物素链亲和素法免疫细胞化学检测耐药相关蛋白P糖蛋白、谷胱甘肽S转移酶π、多药耐药蛋白和拓扑异构酶Ⅱ的表达,逆转录PCR检测多药耐药基因、多药耐药蛋白、拓扑异构酶Ⅱ和谷胱甘肽S转移酶π的表达。结果免疫细胞化学和逆转录PCR显示2组细胞多药耐药蛋白、谷胱甘肽S转移酶π、拓扑异构酶Ⅱ等表达均升高,但体内诱导较体外诱导可以获得更高、更稳定的耐药性。诱导后的Tca8113细胞对氨甲喋呤、卡铂、平阳霉素、长春新碱耐药性明显升高,而对5-氟尿嘧啶耐药性增加不明显。结论体内诱导得到的细胞株耐药性明显高于体外诱导株,肿瘤细胞的耐药现象受多种基因的调节。体内诱导Tca8113卡铂耐药细胞株Tca8113/CBP-vo可以作为肿瘤耐药研究的平台。

Objective To study the drug resistance changes in TcaS113 cell lines by exposing to Carboplatin in the nude mice. Methods The Tca8113 cells were injected subcutaneously in both sides of armpit of nude mice. Two weeks after injection Carboplatin was used subcutaneously around the tumor. Mice were sacrificed about 13 weeks after drug inducing. The sensibility to drugs of the cells was analysed by MTT method. Immunocytochemistry and RT-PCR were utilized to examine the expression of MDR proteins and genes. Results After exposing to Carboplatin, the Tea8113/CBP- vo cells had higher drug-resistance to CBP, MTX, PYM, VCR and higher expression of MRP, GST-π than Tca8113 cells. Conclusion The study showed MDR phenotype was elevated obviously in the inducing group, and in vivo inducing MDR was a good way to establish investigation platform for MDR mechanisms and reversing study.