miR-589-3p/HIF1A轴调控卵巢癌化疗药物的耐药机制

miR-589-3p/HIF1A axis regulates the mechanism of chemotherapeutic drug resistance in ovarian cancer

[目的]探讨miRNA介导的卵巢癌抵抗化疗药物诱导细胞凋亡的潜在机制。[方法]将卵巢癌细胞SK-OV-3进行缺氧或不缺氧处理,随后加入化疗药物OXA或5-Fu,检测SK-OV-3细胞的凋亡水平。通过miRNA高通量测序检测缺氧和不缺氧状态下的差异miRNA。在缺氧状态下的SK-OV-3细胞中过表达差异miRNA后,检测SK-OV-3细胞的凋亡水平。通过miRDB在线分析预测、siRNA敲除实验和荧光素酶报告系统鉴定miRNA的底物。[结果]将SK-OV-3细胞进行缺氧或不缺氧处理并加入化疗药物OXA或5-Fu后,缺氧处理的SK-OV-3细胞的凋亡水平显著下降(P<0.05)。当过表达miR-589-3p时,缺氧状态下SK-OV-3细胞的细胞凋亡水平上升(P<0.05)。miR-589-3p靶向并降解HIF1A。敲低HIF1A时,缺氧状态下SK-OV-3细胞的凋亡水平显著上升(P<0.05)。过表达HIF1A时,缺氧状态下SK-OV-3细胞的凋亡水平显著下降(P<0.05)。[结论]缺氧状态下,SK-OV-3细胞减少了miR-589-3p的表达,同时miR-589-3p的底物HIF1A的表达上升,最终提高了自身的耐药性,减少了细胞凋亡(P<0.05)。

[Objective]To explore the potential mechanism of miRNA-mediated ovarian cancer resistance to chemotherapy drugs-induced apoptosis.[Method]Ovarian cancer cells SK-OV-3 were treated with or without hypoxia,followed by the addition of chemotherapeutic drugs OXA or 5-Fu,and the apoptosis level of ovarian cancer cells SK-OV-3 was detected.Detection of differential miRNAs in hypoxic and non-hypoxic states by miRNA high-throughput sequencing.apoptosis levels of SK-OV-3 cells were detected after overexpression of differential miRNAs in ovarian cancer cells SK-OV-3 under hypoxia.The substrates of miRNAs were identified by miRDB online analysis prediction,siRNA knockdown assay and luciferase reporter system.[Result]After hypoxia or non-hypoxia treatment of SK-OV-3 cells and the addition of chemotherapeutic drugs OXA or 5-Fu,the apoptosis level of hypoxia-treated SK-OV-3 cells decreased significantly(P<0.05).When miR-589-3p was overexpressed,the apoptosis level of SK-OV-3 cells increased under hypoxia(P<0.05).miR-589-3p targets and degrades HIF1A.When HIF1A was knocked down,the apoptosis level of SK-OV-3 cells under hypoxia increased significantly(P<0.05).When HIF1A was overexpressed,the apoptosis level of SK-OV-3 cells under hypoxia decreased significantly(P<0.05).[Conclusion]Under hypoxia,SK-OV-3 cells reduced the expression of miR-589-3p,while the expression of miR-589-3p's substrate HIF1A increased,which ultimately improved their own drug resistance and reduced cell apoptosis(P<0.05).