多药耐药口腔腺样囊性癌细胞株(Acc-3/CDDP)的建立及其细胞中相关耐药基因表达的变化

目的:采用细胞毒性化疗药物顺铂(Cisplatin,CDDP)诱导口腔腺样囊性癌Acc-3细胞系,得到耐药表型的细胞亚系Ace-3/CDDP,检测其多药耐药性、探讨与其亲代Ace-3细胞系的耐药相关基冈变化.方法:用浓度递增法建立Ace-3/CDDP耐药细胞系;用MTT法检测Ace-3/CDDP细胞的耐药指数,RT-PCR检测多药耐药基因MDR-1、MRP-1、GST-π和Bcl-2基因的mRNA表达水平.结果:Acc-3/cDDP对CDDP、5-氟尿嘧啶(5-Fu)、长春新碱(VCR)、氨甲喋呤(MTX)和平阳霉素(PYM)的耐药指数(Resistance factor,RF)分别为8.9、1.97、2.83、2.42、1.1;RT-PCR显示:Aec-3/CDDP中MRP-1、GST-π、Bcl-2较Acc-3升高,Acc-3/CDDP中有MDR-1基因表达,而Acc-3中无MDR-l基因表达.结论:MRP-1、GST-π、Bcl-2基因的表达水平在多药耐药口腔腺样囊性癌Aee-3/CDDP细胞中显著升高,这为进一步探讨肿瘤多药耐药的形成机制提供了科学基础.

Establishment and drug-resistance gene expression of cisplatin-resistant cell line Acc-3/CDDP

Objective: To establish a CDDP-resistant cell line from human oral adenoid cystic carcinoma cell line Ace-3 and compare the drag-resistance gene expression between them. Method: The CDDP-resistant cell line Ace-3/CDDP was established by increasing the concentration of CDDP added to Ace-3 cells step wisely. The index of drug-resistance was estimated by MTT assay. The expression of MDR-1,MRP-l,GST-π,and Bcl-2 were measured by RT-PCR optical density OD.Result: The resistance factors (RF=IC_(50)(Acc-3/CDDP)/IC_(50)(Acc-3))for CDDP,5-Fu,VCR,MTX and PYM were 8.9,1.97,2.83,2.42,1.1 respectively. The expression of drug-resistance gene MRP-1,GST-π and Bcl-2 increased in Acc-3/CDDP cells.MDR-1 was expressed in Acc-3/CDDP,but none in Acc-3.Conclusion: MDR-1,MRP-1,GST-πand Bcl-2 were significantly over expressed in CDDP-resistant cell line Ace-3/CDDP,which can provide effective models for the study on multi-drug resistance of oral carcinoma.