BubR1在乳腺癌细胞发生紫杉醇耐药中的作用

Role of BubR1 in Drug Resistance of Breast Cancer Cells to Paclitaxel

目的探讨有丝分裂期检查点蛋白BubR1在乳腺癌细胞发生紫杉醇耐药中的作用。方法设计并合成针对BubR1基因的特异性干扰RNA(siRNA)片段,连接入穿梭质粒后,包装成腺病毒(Ad-BubR1-siRNA),感染MCF-7细胞,采用实时荧光定量RT-PCR和Western blot法评价BubR1干扰对MCF-7细胞BubR1基因和蛋白表达的影响。分别通过cck8试验、染色体计数分析、细胞集落生成试验、Hochest染色和细胞划痕试验评价干扰BubR1后,MCF-7细胞在紫杉醇作用下增殖、染色体稳定性、凋亡及迁移能力等生物学功能。结果重组腺病毒的滴度为1010 pfu/ml,其能有效抑制BubR1基因在mRNA和蛋白水平的表达。干扰BubR1表达后,MCF-7细胞在紫杉醇作用下表现为增殖和迁移能力增强,染色体不稳定性增加,集落生成能力增强,凋亡能力下降。结论下调BubR1基因可能导致乳腺癌细胞MCF-7对紫杉醇的耐药性增强。

Objective To investigate the role of spindle assemble checkpoint protein BubR1 in drug resistanceof breast cancer cells to paclitaxel（PTX）.Methods Specific small interfering RNA（siRNA） fragment to BubR1 gene was designed and synthesized,then inserted into shuttle plasmid and packaged into adenovirus（Ad-BubR1-siRNA）.MCF-7 cells were infected with Ad-BubR1-siRNA,based on which the effect on expression of BubR1 was evaluated by real-time fluorescent quantitative RT-PCR and Western blot.The biological function of MCF-7 cells after BubR1 interference and treatment with PTX,such as proliferation,chromosome stability,apoptosis and migration ability,were evaluated by cck8 test,chromosome counting,cell colony formation test,Hochest staining and cell scarification test.Results Recombinant adenovirus Ad-BubR1-siRNA,at a titer of 1010 pfu / ml,inhibited the expression of BubR1 at mRNA and protein levels effectively.After interference of BubR1 expression,the proliferation and migration abilities as well as chromosome instability of MCF-7 cells treated with PTX increased,while the apoptosis ability decreased.Conclusion The down-regulation of BubR1 gene might increase the resistance of breast cancer MCF-7 cells to PTX.