人舌鳞状细胞癌顺铂耐药细胞系的建立及其耐药基因表达

目的建立耐顺铂(cisplatin,CDDP)人舌鳞状细胞癌细胞系Tca8113/CDDP,探讨其与亲代细胞耐药基因表达水平的差异。方法以人舌鳞癌Tca8113细胞系为实验对象,采用间歇性加药,逐步递增剂量,体外连续诱导培养,诱导其耐药性。用MTT法测定细胞的耐药指数和IC50。RT-PCR检测MDR-1、MRP、GST-π和Bcl-2的表达状况,测其光密度比值。结果建立顺铂耐药细胞系Tca8113/CDDP,耐药指数为9.2倍。RT-PCR显示:其MDR-1、MRP、GST-π和Bcl-2的表达均升高,光密度比值MRP:Tca8113/CDDP∶Tca8113=6.667∶1.111=6倍,Bcl:Tca8113/CDDP∶Tca8113=0.912∶0.549=1.6倍,GST-π∶Tca8113/CDDP∶Tca8113=2∶1=2倍。结论耐CDDP人舌鳞状细胞癌细胞系Tca8113/CDDP有多种耐药基因的表达上升,提示肿瘤细胞的耐药现象受到多种基因的调节。

Establishment and Drug-Resistance Gene Expression of Cisplatin-Resistant Cell Line Tca8113/CDDP

OBJECTIVE: To establish a CDDP-resistant cell line from human tongue squamous cell carcinoma cell line Tca8113 and to compare the drug-resistance gene expression between them. METHODS: The concentration of CDDP added to Tca8113 cells was increased stepwise to induce the CDDP-resistance. The index of drug-resistance was estimated by MTT assay. The expression of MDR-1, MRP, GST-pi and Bcl-2 was measured by RT-PCR optical density OD. RESULTS: The CDDP-resistant cell line, Tca8113/CDDP was established, whose drug-resistance index was 9.2. The expression of MDR-1, MRP, cyclin D1 and Bcl-2 increased respectively. CONCLUSION: The expression of drug-resistance gene in Tca8113/CDDP was significantly higher than that in Tca8113.