人卵巢癌顺铂耐药细胞株OVCAR-3/DDP的建立及相关基因表达水平的研究

ESTABLISHMENT OF CISPLATIN-RESISTANT HUMAN OVARIAN CELL LINE OVCAR-3/DDP AND DETECTION OF RELATED GENE EXPRESSION LEVEL

目的建立人卵巢癌顺铂耐药细胞株OVCAR-3/DDP,初步探讨其耐药机制。方法采用顺铂(DDP)浓度梯度递增诱导法,建立人卵巢癌OVCAR-3细胞株的顺铂耐药细胞株OVCAR-3/DDP;通过细胞计数和MTT法评估细胞增殖情况,流式细胞仪检测细胞周期情况,以评价OVCAR-3/DDP的生物学特性;采用RT-PCR法检测Mfn2、EGFR及MMP2 mRNA水平。结果成功建立了OVCAR-3/DDP顺铂耐药细胞株,其对DDP耐药指数是1.92;与亲本细胞株OVCAR-3相比,OVCAR-3/DDP生长速度减慢,倍增期是OVCAR-3细胞的1.36倍(P<0.05);流式细胞术检测结果显示细胞主要停滞于G2/M期(P<0.05);与OVCAR-3细胞相比,OVCAR-3/DDP细胞中Mfn2、EGFR及MMP2 mRNA水平均明显升高(P<0.05)。结论建立的OVCAR-3/DDP细胞株对顺铂耐药性稳定;OVCAR-3/DDP细胞对顺铂耐药可能与Mfn2、EGFR、MMP2基因转录水平上调有关。

Objectives To establish cisplatin-resistant human ovarian cancer cell line OVCAR-3/DDP, and discuss its resistance mechanism.Methods Stepwise selection in increasing concentration of cisplatin was adopted to establish cisplatin-resistant OVCAR-3/DDP cell line.Cell count and MTT method were used to evaluate cell proliferation;Flow cytometry was used to detect the cell cycle;RT-PCR was used to analyze the mRNA levels of Mfn2,EGFR and MMP2.Results Cisplatin-resistance cell line OVCAR-3/DDP was successfully established,DDP resistance index of which was 1.92;Cell growth of OVCAR-3/DDP cells was slow,its doubling time was 1.36-fold longer than that of OVCAR-3 cells （P 〈0.05）;Flow cytometry results show that the cells were main stagnation in G2/M phase （P 〈0.05）;The mRNA levels of Mfn2,EGFR and MMP2 were increased significantly in OVCAR-3/DDP compared with those in OVCAR-3 cells （P 〈0.05）.Conclusion The cisplatin resistance of OVCAR-3/DDP cell line is stable, which probably is related to the increased expression of Mfn2,EGFR and MMP2 genes.