IL-17A对卵巢癌顺铂耐药的影响及其机制的研究

Study of the effects and underlying mechanisms of IL-17A on the cisplatin-based resistance of ovarian cancer

目的:探究IL-17A对卵巢癌(OVCA)顺铂(DDP)耐药的影响及其可能的作用机制。方法:应用MTT法检测IL-17A(0.1、1、10、100 ng/m L,处理12、24、48、72 h)对A2780(顺铂敏感卵巢癌细胞株)和OVCAR3(顺铂耐药卵巢癌细胞株)增殖的影响;应用MTT法检测IL-17A(1 ng/m L,处理24 h)对A2780和OVCAR3细胞DDP耐药的影响,并以rh IL-17RA中和抗体(IL-17RA m Ab)和Gli1抑制剂Gant61进行相应的阻断实验;应用Western blotting法检测IL-17A处理A2780和OVCAR3细胞后,细胞中耐药相关分子ABCG2、MDR1和Hedgehog(Hh)信号通路核转录因子Gli1的蛋白表达,并分别以IL-17RA m Ab和Gant61进行相应的阻断实验。结果:IL-17A对A2780和OVCAR3的细胞增殖无显著性影响,但可显著增加DDP降低的A2780和OVCAR3细胞活性(P<0.05),分别以IL-17RA m Ab和Gant61进行阻断实验均可消除由IL-17A加剧的A2780和OVCAR3细胞DDP的耐药性;IL-17A可上调A2780和OVCAR3细胞耐药相关分子ABCG2,MDR1和Gli1的蛋白表达,分别以IL-17RA m Ab和Gant61进行阻断实验均可抑制IL-17A引起的A2780和OVCAR3细胞中ABCG2,MDR1和Gli1蛋白表达。结论:IL-17A可作用于IL-17RA并激活Gli1介导的Hh信号通路,进而促进耐药相关分子ABCG2和MDR1表达,从而加剧以DDP为基础的OVCA化疗耐药性。

Objective： To study the effects and underlying mechanisms of IL-17 A on the cisplatin-based resistance of ovarian cancer.Methods：MTT assay was used to detect the effect of IL-17 A on cell growth, A2780 and OVCAR3 cells were treated with 0, 0.1, 1, 10 or100 ng/m L IL-17 A for 12, 24, 48 or 72 h. MTT assay was applied to detect the effect of viability of IL-17A（1ng/m L, 24 h） on A2780 and OVCAR3 cells treated with DDP（A2780, 10 μmol/L; OVCAR3, 100 μmol/L）, and the neutralizing monoclonal antibody against IL-17 A receptor（IL-17 RA m Ab） and the widely used chemical inhibitor for Gli1（Gant61） were used to do the blocking test respectively. Western blotting assay was used carried out to detect the protein levels of ABCG2, MDR1 and Gli1 on IL-17 A treated A2780 and OVCAR3 cells,and then neutralizing IL-17 RA m Ab and Gant61 were obtained to do the blocking test respectively. Results：IL-17 A alone had no effect on the viability of A2780 and OVCAR3 cells, but it could increase the viability of A2780 and OVCAR3 cells with DDP treatment（P〈0.05）.Furthermore, neutralizing IL-17 RA m Ab and Gant 61 could respectively block the above effect of IL-17 A on DDP-sensitivity significantly.IL-17 A could increase the expressions of ABCG2, MDR1 and Gli1 in A2780 and OVCAR3 cells. Furthermore, neutralizing IL-17 RA m Ab and Gant 61 could respectively block the enhanced effects of IL-17 A on ABCG2, MDR1 and Gli1 levels in A2780 and OVCAR3 cells.Conclusion：IL-17 A can exacerbate cisplatin-based resistance of ovarian cancer cells via up-regulating the expression of ABCG2 and MDR1,partly through IL-17RA-Gli1-mediated Hh signal pathway, which may provide a novel strategy to improve the chemo-resistance of OVCA.