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作 者:潘洁雪[1] 林凤[2] PAN Jiexue;LIN Feng(Reproductive Medicine Center, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015;Department of Gynecology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015)
机构地区:[1]温州医科大学附属第一医院生殖医学中心,浙江温州325015 [2]温州医科大学附属第一医院妇科,浙江温州325015
出 处:《温州医科大学学报》2017年第11期805-809,共5页Journal of Wenzhou Medical University
基 金:浙江省中医药管理局青年人才基金项目(2017ZQ020);浙江省医药卫生科技计划项目(2017KY462);温州市科技局科研基金资助项目(Y20160037)
摘 要:目的:探索上皮性卵巢癌细胞卡铂耐药的机制,以提高卵巢癌患者化疗的疗效。方法:应用CCK8法检测卡铂处理后的上皮性卵巢癌细胞增殖,Western blot法检测上皮间质化转变相关蛋白E-Cadherin和Vimentin表达情况,流式细胞技术检测细胞表面CD133的变化情况,使用si RNA敲降AMP激活的蛋白激酶家族成员5(ARK5)后再观察卡铂的作用。结果:上皮性卵巢癌细胞系SKOV3与A2780的卡铂IC50分别为2.824 μg/m L 与1.760 μg/m L,并且卡铂能诱导上皮性卵巢癌细胞发生上皮间质化转变;卡铂诱导ARK5显著上调,而不影响SNF1/AMP激活的蛋白激酶表达,敲降ARK5基因能显著减弱卡铂诱导卵巢癌细胞产生上皮间质化转变的效果。结论:ARK5在上皮性卵巢癌细胞卡铂耐药机制中具有重要作用,降低ARK5的表达可以减弱卡铂诱导卵巢癌细胞上皮间质化转变的作用。Objective: To investigate the mechanisms of carboplatin resistance in epithelial ovarian cancer (EOC) cells, and provide preliminary exploration on improving the effect of chemotherapy in ovarian cancer patients. Methods: Cell counting kit-8 was used to evaluate the effect of carboplatin on the proliferation of SKOV3 and A2780 cells, meanwhile the expression of epithelial-mesenchymal transition related proteins E-cadherin and vimentin were detected by Western blot. Flow cytometry assay was used to detect the expression of CD133. In addition, knockdown of ARK5 was to evaluate the role of ARK5 in the carboplatin-induced epithelial-mesenchymal transition. Results: The IC50 of carboplatin in SKOV3 and A2780 was 2.824 μg/mL and 1.760 μg/mL, respectively. Furthermore, we also found that carboplatin could significantly induce epithelial-mesenchymal transition in EOC cells. Carboplatin could increase the expression of ARK5, but not SNARK. Knockdown of ARK5 could attenuate the induction of epithelial-mesenchymal transition. Conclusion: ARK5 plays an important role in carboplatin resistance in EOC cells. Knockdown of ARK5 may reduce the carboplatin-induced epithelial-mesenchymal transition.
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