Twist表达增强乳腺癌MCF-7细胞的多药耐药性

Enhanced multidrug resistance by Twist stable expression in MCF-7 cell line of human breast cancer

目的探讨MCF-7/Twist稳定表达细胞株的多药耐药性。方法 MCF-7细胞中转染Twist,G418筛选,建立稳定表达细胞株MCF-7/Twist,Western blotting法检测Twist表达;观察增殖状态;MTT法测阿霉素、紫杉醇、长春新碱和羟喜树碱对细胞株增殖活性的影响,分析稳定转染细胞株对药物的敏感性;应用RT-PCR及Western blotting方法,检测耐药相关糖蛋白P-gp和乳腺癌耐药蛋白BCRP转录水平和蛋白水平的变化。结果 G418筛选获得稳定表达Twist的细胞株,对阿霉素、紫杉醇、长春新碱和羟喜树碱具有明显耐药性,耐药相关分子P-gp和BCRP转录和表达明显升高。结论 Twist表达增强乳腺癌细胞的多药耐药性,多药耐药性相关分子的转录和表达升高与耐药性增强现象一致。

Objective To study multidrug resistance （MDR） enhanced by Twist stable expression in MCF-7 cell line of human breast cancer in vitro. Methods Twist gene was introduced into MCF-7 to establish a stable expression of the cell line MCF-7/Twist. The expression of Twist was detected with Western blotting. The cell proliferation and viability were observed under microscope and measured with MTT assay. Reverse transcription- polymerase chain reaction was used to test transcriptional level of genes involved in MDR. The relevant protein expression level of P-gp and breast cancer resistance protein （BCRP） was detected with Western blotting. Results The results showed low sensitivity of MCF-7/Twist to several chemotherapeutic drugs including adrimycin, taxol, vincristine and hydroxycamptothecin. In addition, MCF-7/Twist, the stable expression cell line, was generated through G418 selection, which was endowed with MDR in responsible to the above drugs. Conclusion The expression of Twist enhances MDR, which accords with the level of transcription and expression of P-gp and BCRP.