MMP2调节高级别浆液性卵巢癌细胞对PARP抑制剂Olaparib敏感性的机制研究OA
Effect of MMP2 Expression on Cell Sensitivity to PARP Inhibitor Olaparib in High-grade Serous Ovarian Cancer Cells
目的 探讨基质金属蛋白酶 2(matrix metalloproteinase-2,MMP2)对高级别浆液性卵巢癌(high-grade serous ovarian cancer,HGSOC)细胞聚-ADP核糖聚合酶抑制剂(poly-ADP ribose polymerase in-hibitor,PARPi)敏感性的影响及机制.方法 培养 HGSOC 细胞株 HeyA8,分为对照组、sh-NC 组、sh-MMP2 组,RT-qPCR和蛋白质印迹检测MMP2 表达,CCK-8 法检测不同浓度奥拉帕尼(olaparib)处理下细胞存活率.再将HeyA8 细胞分为sh-NC组、sh-MMP2 组、sh-NC+PARPi组、sh-MMP2+PARPi组,细胞划痕实验、Annexin V-FITC/PI法、Hochest 33258 染色分别检测细胞迁移与凋亡,蛋白质印迹测定血管内皮生长因子(vascular endothelial growth factor,VEGF)/磷脂酰肌醇 3-激酶(phosphoinositide 3-kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)信号通路相关蛋白表达.结果 与对照组、sh-NC组比较,sh-MMP2组HeyA8 细胞中MMP2 mRNA及蛋白相对表达量下调,Olaparib处理后的细胞存活率降低(P<0.05).与sh-NC组比较,sh-MMP2 组细胞划痕愈合率降低,凋亡率增加,可见核浓缩、碎裂的凋亡形态,血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)、血管内皮细胞生长因子受体2(vascular endothe-lial growth factor receptor 2,VEGFR2)、磷酸化PI3K(Tyr458)[p-PI3K(Tyr458)]、磷酸化Akt(Ser473)[p-Akt(Ser473)]蛋白相对表达水平均下调(P<0.05);与sh-NC+PARPi组比较,sh-MMP2+PARPi组细胞划痕愈合率降低,凋亡率增加,核浓缩、碎裂的凋亡细胞更多,VEGFA、VEGFR2、p-PI3K(Tyr458)、p-Akt(Ser473)蛋白相对表达水平也均下调(P<0.05).结论 沉默MMP2 表达能够促进PARPi诱导的HGSOC细胞凋亡,增加PARPi敏感性.
Objective To investigate the effect of matrix metalloproteinase 2(MMP2)on the sensitivity of poly-ADP ribose polymerase inhibitor(PARPi)in high-grade serous ovarian cancer(HGSOC)cells and the underlying mechanism.Methods The HGSOC cell line HeyA8 cells were divided into 3 groups:control group,sh-NC group,sh-MMP2 group.RT-qPCR and Western blot-ting were used to detect the expression of MMP2.CCK-8 method was used to detect the cell survival rate under different concentrations of Olaparib.Then HeyA8 cells were divided into 4 groups:sh-NC group,sh-MMP2 group,sh-NC+PARPi group,sh-MMP2+PARPi group.Wound-healing assay,Annexin V-FITC/PI method and Hochest 33258 staining were used to detect cell migration and ap-optosis,respectively.Western blotting was used to determine the expression levels of proteins related to the vascular endothelial growth factor(VEGF)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt).Results Compared with those in the control group and the sh-NC group,the relative expression levels of MMP2 mRNA and protein of HeyA8 cells in the sh-MMP2 group were downregulated,and the cell survival rate after Olaparib treatment was reduced(P<0.05).Com-pared with those in the sh-NC group,the wound-healing rate of HeyA8 cells in the sh-MMP2 group was decreased,the apoptosis rate was increased,nuclear condensation and fragmentation of apop-totic cells were observed,the relative protein expression levels of vascular endothelial growth factor A(VEGFA),vascular endothelial growth factor receptor 2(VEGFR-2),phosphorylated PI3K(Tyr458)[p-PI3K(Tyr458)]and phosphorylated Akt(Ser473)[p-Akt(Ser473)]were all downregulated(P<0.05).Compared with those in the sh-NC+PARPi group,the wound-healing rate of HeyA8 cells in the sh-MMP2+PARPi group was decreased,the apoptosis rate was in-creased,more apoptotic cells with nuclear condensation and fragmentation were observed,the rela-tive protein expression levels of VEGFA,VEGFR2,p-PI3K(Tyr458)and p-Akt(Ser473)were downregulated(P<0.05).Conclusion Silencing MMP2 expression can promote PARPi-induced apoptosis in HGSOC cells,and increase PARPi sensitivity.
邵萍;邓青春
海南医科大学第二附属医院妇科 海口市,570102海南医科大学第二附属医院妇科 海口市,570102
医药卫生
高级别浆液性卵巢癌基质金属蛋白酶2奥拉帕尼药物敏感性血管内皮生长因子/磷脂酰肌醇3-激酶/蛋白激酶B通路
high-grade serous ovarian cancermatrix metalloproteinase 2olaparibdrug sensitivityvascular endothelial growth factor/phosphatidylinositol 3-kinase/pro-tein kinase B pathway
《医学分子生物学杂志》 2026 (2)
164-170,7
海南省自然科学基金高层次人才项目(No.821RC712) This work was supported by a grant from the High-level Talent Program of Natural Science Foundation in Hunan(No.821RC712)
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