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目的探讨髓样分化因子(myeloid differentiation factor88,MyD88)参与调节卵巢癌细胞耐药性的具体机制。方法在卵巢癌耐药细胞株A2780/Taxol中采用特异性小干扰RNA(short interfering RNA,siRNA)抑制MyD88的表达,通过CCK-8试剂盒检测细胞增殖情况以及细胞在紫杉醇处理后的存活情况。Western blot检测磷酸化蛋白激酶B(p-Akt)、X-连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)以及多药耐药蛋白(multidrug resistance 1,MDR1)的表达情况。进一步在A2780/Taxol细胞中通过LY294002抑制PKB/Akt信号通路,通过小干扰RNA抑制MDR1的表达,CCK-8试剂盒检测细胞在紫杉醇处理后的存活情况。结果在A2780/Taxol细胞株中抑制MyD88的表达,细胞的增殖速度明显低于对照组(P<0.05)。A2780/Taxol细胞对紫杉醇的耐药性也明显降低,半数抑制浓度(IC50)降低至对照组的50%(P<0.01),且p-Akt、XIAP和MDR1在药物刺激下的升高也被明显抑制。同时抑制p-Akt通路的活性或抑制MDR1的表达后A2780/Taxol细胞的耐药性也出现明显下降,IC50分别降低至对照组的80%和50%,且差异具有统计学意义(P<0.05)。结论 MyD88蛋白通过调节PKB/Akt信号通路、XIAP抗凋亡蛋白以及MDR1的表达参与卵巢癌细胞对紫杉醇的耐药性。通过抑制MyD88可望改善卵巢癌治疗中的耐药现状。
Objective To investigate the mechanism of myeloid differentiation factor (MyD88) involved in the regulation of drug resistance in ovarian cancer cells. Methods The expression of MyD88 was inhibited by short interfering RNA (siRNA) in A2780 / Taxol resistant ovarian cancer cell line. The proliferation of cells and the survival of cells after paclitaxel treatment were detected by CCK-8 kit. Western blot was used to detect the expression of phosphorylated protein kinase B (p-Akt), X-linked inhibitor of apoptosis protein (XIAP) and multidrug resistance protein 1 (MDR1) Further, the PKB / Akt signaling pathway was inhibited by LY294002 in A2780 / Taxol cells and the expression of MDR1 was inhibited by small interfering RNA. The survival of cells after paclitaxel treatment was examined by CCK-8 kit. Results The expression of MyD88 was inhibited in A2780 / Taxol cell lines, and the proliferation rate of cells was significantly lower than that of the control (P <0.05). The drug resistance of paclitaxel in A2780 / Taxol cells was also significantly reduced, and the IC50 decreased to 50% (P <0.01), and the increase of p-Akt, XIAP and MDR1 under drug stimulation Obvious inhibition. A2780 / Taxol cells also showed a significant decrease in drug resistance, IC50 were reduced to 80% and 50% of the control group, respectively, and the difference was statistically significant (P <0.05 ). Conclusion MyD88 protein is involved in the resistance of paclitaxel to ovarian cancer cells by regulating the expression of PKA / Akt signaling pathway, XIAP anti-apoptotic protein and MDR1. By inhibiting MyD88 is expected to improve the treatment of ovarian cancer drug resistance status.