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XIAP基因与耐紫杉醇卵巢癌细胞A2780/Taxol耐药关系的研究
Study on the Relationship Between XIAP Gene and Resistance of Taxol in Ovarian Cancer
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摘要: 目的 探讨X连锁凋亡抑制蛋白（XIAP）基因与卵巢癌对紫杉醇耐药的相关性。方法 以5、10、20、40、80、160、320 ng/mL的紫杉醇处理卵巢癌紫杉醇敏感细胞A2780和耐药细胞A2780/T,噻唑蓝（MTT）比色法检测各组细胞的抑制率（IR）, 逆转录实时荧光定量PCR（RT-qPCR）、蛋白质印记（Western blot）检测紫杉醇100 ng/mL处理A2780和A2780/T细胞后XIAP基因和蛋白的表达；将A2780/T细胞分为空转染组（转染空载体质粒）、小干扰RNA（siRNA）-XIAP（siRNA-XIAP）组（转染siRNA-XIAP质粒）、非特异性转染组（转染非特异性质粒）、空白对照组（不转染质粒），RT-qPCR和Western blot检测各组细胞XIAP基因和蛋白的表达，并加入含不同质量浓度的紫杉醇（0、1 000、1 500、2 000、2 500 ng/mL），流式细胞仪检测细胞凋亡率。结果 不同质量浓度紫杉醇处理后，A2780细胞IR随紫杉醇质量浓度的增加而增高（P<0.05），A2780/T细胞IR无明显变化。紫杉醇100 ng/mL处理A2780和A2780/T细胞后，A2780细胞XIAP mRNA的表达低于紫杉醇未处理组（P<0.05），A2780/T细胞中XIAP mRNA的表达与紫杉醇未处理组差异无统计学意义(P>0.05)，但A2780/T细胞XIAP mRNA和蛋白表达均高于A2780紫杉醇处理组（P均<0.05）；siRNA-XIAP组XIAP mRNA和蛋白的表达均低于其他各组（P<0.05），在紫杉醇2 000 ng/mL及2 500 ng/mL作用下，siRNA-XIAP组凋亡率高于其他各组（P<0.05）。结论 卵巢癌对紫杉醇耐药与XIAP基因的高表达有关，特异性的siRNA可通过降低XIAP的表达，促进细胞凋亡，增加耐药癌细胞对紫杉醇的敏感性。Abstract: Objective To research the expression of X-linked inhibitor of apoptosis protein gene （XIAP） on paclitaxel resistance in ovarian cancer. Methods A2780 and A2780/T cells were treated with paclitaxel respectively at the concentrations of 5 ng/mL, 10 ng/mL, 20 ng/mL ,40 ng/mL, 80 ng/mL, 160 ng/mL, 320 ng/mL, then the inhibition rate of cells were detected by MTT assay. The expression of XIAP mRNA and protein among the A2780 and A2780/T cells treated respectively with paclitaxel at the concentration of 100 ng/mL was detected by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and Western blot. The A2780/T cells were divided into blank group, empty group, small interfering RNA (siRNA) XIAP group and siRNA-non-specific group. The expression of XIAP mRNA and protein of four groups were detected by RT-qPCR and Western blot. Apoptotic rate of these groups with addition of paclitaxel at the concentrations of 0 ng/mL, 1 000 ng/mL, 1 500 ng/mL, 2 000 ng/mL and 2 500 ng/mL were detected by flow cytometry. Results After the treatments on A2780 and A2780/T cells with the different concentrations of paclitaxel, the inhibition rate of A2780 cells were gradually increased with the increased paclitaxel concentrations (P<0.05), while there were no obvious differences in A2780/T cells (P>0.05). After the treatment on these cells with paclitaxel at the concentration of 100 ng/mL, the expression of XIAP mRNA was lower than that non-treatment with paclitaxelin A2780 cells (P<0.05), and the expression of XIAP mRNA in the A2780/T cells were no statistical significance between the treatment group and non-treatment group with paclitaxel (P>0.05). However, the expression of A2780/T cells’XIAP mRNA and protein treated with paclitaxel were higher than A2780 cells’ (P<0.05). The expression of XIAP mRNA and protein in siRNA-XIAP group was lower than those of other groups (P<0.05). The apoptotic rateof siRNA-XIAP group was higher than those of other groups treated with the paclitaxel at concentrations of 2 000 ng/mL and 2 500 ng/mL (P<0.05). Conclusion XIAP’s high expression on mRNA and protein was correlated with ovarian cancer paclitaxel-resistance, specific siRNA can promote cell apoptosis by reducing the expression of XIAP, and increase the sensitivity of drug-resistant cancer cells to paclitaxel.
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期刊类型引用(9)
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