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目的探讨卵巢癌细胞系A2780紫杉醇(PTX)体外化疗后细胞色素P4501B1(CYP1B1)表达的变化,研究CYP1B1基因表达与肿瘤细胞耐药的关系。方法2004年天津医科大学总医院采用四甲基偶氮唑蓝(MTT)比色法观察不同质量浓度PTX(50、25、12.5、6.25、3.13、1.56、0.78mg/L)对A2780细胞体外生长的抑制作用。应用RT-PCR技术检测5mg/L PTX分别处理A2780细胞24h、48h、72h和50mg/L PTX处理A2780细胞24h后存活的卵巢癌细胞中CYP1B1 mRNA的表达。Western blot检测5mg/L PTX处理不同时间后和50mg/L PTX处理A2780细胞后CYP1B1蛋白表达。结果PTX抑制卵巢癌细胞生长,不同浓度PTX作用A2780细胞72h后的抑制率分别为90.45%、78.63%、66.77%、58.38%、45.90%、36.41%、15.38%,随药物质量浓度的下降,细胞受抑制程度明显降低(P<0.05)。PTX处理后存活的卵巢癌细胞中CYP1B1 mRNA及蛋白的表达量增加,高于未加药的对照组;5mg/L处理48h、72h和50mg/L组CYP1B1的表达量高于5mg/L处理24h组(P<0.05)。结论CYP1B1基因在卵巢癌细胞系中呈高表达,在卵巢癌细胞系A2780体外化疗中起抑制作用。
Objective To investigate the change of cytochrome P4501B1 (CYP1B1) expression after chemotherapy in vitro in ovarian cancer cell line A2780 paclitaxel (PTX) and to study the relationship between CYP1B1 gene expression and drug resistance in tumor cells. Methods In 2004, MTT colorimetry was used to observe the growth of A2780 cells in vitro by different concentrations of PTX (50,25,12.5,6.25,3.13,1.56,0.78mg / L) Inhibition. RT-PCR was used to detect the expression of CYP1B1 mRNA in surviving ovarian cancer cells treated with 5 mg / L PTX for 24 h, 48 h, 72 h and 50 mg / L PTX, respectively. The expression of CYP1B1 protein in A2780 cells treated with 5 mg / L PTX at different time and 50 mg / L PTX was detected by Western blot. Results PTX inhibited the growth of ovarian cancer cells. The inhibitory rates of PTX treated with different concentrations of PTX for 72 hours were 90.45%, 78.63%, 66.77%, 58.38%, 45.90%, 36.41% and 15.38%, respectively. With the decrease of drug concentration, Inhibition was significantly reduced (P <0.05). The expression level of CYP1B1 mRNA and protein in ovarian cancer cells survived after PTX treatment was higher than that of the untreated control group. The expression level of CYP1B1 in 5mg / L treatment group was higher than that in 5mg / L treatment group at 24h, 72h and 50mg / L Group (P <0.05). Conclusion The CYP1B1 gene is highly expressed in ovarian cancer cell lines and plays an inhibitory role in the chemotherapy of ovarian cancer cell line A2780 in vitro.