摘要
目的应用人乳腺癌细胞制备紫杉醇耐药细胞模型,探讨紫杉醇对乳腺癌细胞中紫杉醇耐药基因Txr1表达的影响。方法比较紫杉醇预处理前后不同浓度紫杉醇对乳腺癌细胞的增殖和细胞周期的改变;用RT-PCR方法检测Txr1、TSP1和MDR1的mRNA水平变化;用Western blot检测Txr1和TSP1的蛋白质水平的变化。结果紫杉醇预处理后,紫杉醇抑制乳腺癌细胞(MCF-7细胞)的生长作用明显减弱。流式细胞仪分析表明,MCF-7细胞经过紫杉醇作用后被阻滞于细胞周期的G2M期。紫杉醇作用于MCF-7细胞后,Txr1的mRNA水平上调,TSP1下调,而MDR1表达无明显改变。类似的结果在蛋白的水平被Western blot进一步证实。结论紫杉醇可能通过Txr1上调抑制TSP1的表达从而诱导肿瘤细胞的耐药性。
Objective To investigate the effect of Taxol resistance gene 1（Txr1） on Taxol-resistance in human breast cancer cell line in vitro. Methods Human breast cancer cell line,MCF-7 was pre-treated with Taxol,and then cell proliferation and cell cycle of MCF-7 were respectively analyzed by MTT assay and flow cytometry in response to different concentration of Taxol. The expression of Txr1,TSP1 and MDR1 in mRNA and protein levels were detected by RT-PCR and western blot,respectively. Results In response to Taxol,the inhibition of MCF-7 proliferation was reduced in Taxol pre-treated group as compared with the control group. The flow cytometry analysis indicated that MCF-7 cells were arrested at G2M phase. After Taxol pre-treatment,the expression of Txr1 mRNA are increase,expression of TSP1 mRNA was decrease,while expression of MDR1 mRNA was no significant change in MCF-7 as compared with the control. The similar changes of Txr1,TSP1 and MDR1 were also conformed in MCF-7 by western blot in protein level after Taxol pre-treatment. Conclusion Taxol may induce taxol-resistance in cancer cells by inhibiting TSP1 expression via up-regulating of Txr1.
出处
《中国医药生物技术》
CSCD
2010年第6期429-433,共5页
Chinese Medicinal Biotechnology
