PROM2调控卵巢癌细胞增殖、迁移和凋亡的机制研究

Mechanism of PROM2 in regulating the proliferation,migration,and apoptosis of ovarian cancer cells

目的探讨PROM2基因下调在卵巢癌中的作用机制及其与卵巢癌细胞增殖、迁移侵袭和凋亡的关系。方法利用GEPIA数据库、TCGA数据库、Ualcan数据库分析PROM2于卵巢癌和正常组织中的表达差异以及相关临床意义。合成siRNA敲低SKOV3和ES-2细胞中PROM2的表达,采用CCK-8实验、平板克隆和划痕法检测细胞增殖和迁移,Transwell实验观察细胞迁移和侵袭能力,流式细胞术检测细胞凋亡,以验证PROM2对卵巢癌细胞功能的影响。Western blot检测敲低PROM2对上皮间质转化(EMT)相关蛋白、信号通路蛋白及凋亡相关蛋白的影响。结果敲低PROM2后,与si-NC#组相比,si-PROM2#1组和si-PROM2#2组的SKOV3和ES-2人卵巢癌细胞的增殖、迁移、侵袭、克隆能力均明显减弱,而细胞凋亡率明显增加,差异有统计学意义(P<0.05);Western blot检测结果显示,si-PROM2#1组和si-PROM2#2组与si-NC#组比较,凋亡相关蛋白Bcl-2表达降低,Bax表达升高,上皮间质转化(EMT)相关蛋白E-cadherin表达升高,N-cadherin和Vimentin表达降低,差异均有统计学意义(P<0.05);WNT/β-catenin信号转导通路相关蛋白β-catenin、TCF-4、c-myc及CyclinD1表达均降低,差异有统计学意义(P<0.05)。结论PROM2敲低可抑制SKOV3和ES-2人卵巢癌细胞的增殖侵袭、迁移以及卵巢癌上皮间质转化,促进细胞凋亡,其机制可能与WNT/β-catenin信号转导通路受到抑制有关。

Objective To investigate the mechanism of PROM2 gene down-regulation in ovarian cancer and its relationship with proliferation,migration,invasion,and apoptosis of ovarian cancer cells.Methods Differential expression of PROM2 in ovarian cancer and normal tissues and its clinical significance were analyzed using the GEPIA database,TCGA database,and Ualcan database.siRNA was synthesized to knock down the expression of PROM2 in SKOV3 and ES-2 cells.CCK-8 assay,plate cloning,and scratch assay were used to detect cell proliferation and migration,Transwell assay to observe the cell migration and invasion ability,and flow cytometry to detect apoptosis,in order to verify the effect of PROM2 on the function of ovarian cancer cells.Effects of knockdown of PROM2 on epithelial mesenchymal transition(EMT)-related proteins,signaling pathway proteins,and apoptosis-related proteins were analyzed using Western blot.Results After knocking down PROM2,the proliferation,migration,invasion,and clonogenic ability of SKOV3 and ES-2 human ovarian cancer cells in the si-PROM2#1 and si-PROM2#2 groups were significantly reduced,while the apoptosis rate was significantly increased,compared with those in the si-NC#group(P<0.05).Western blot results showed that si-PROM2#1 and si-PROM2#2 groups,compared with the si-NC#group,showed decreased expression of apoptosis-related protein Bcl-2,increased expression of Bax,increased expression of EMT-related protein E-cadherin,and decreased expression of N-cadherin and Vimentin,and the differences were all statistically significant(P<0.05).WNT/β-catenin signaling pathway-related proteinsβ-catenin,TCF-4,c-myc,and CyclinD1 expression were all reduced,and the differences were all statistically significant(P<0.05).Conclusion PROM2 knockdown inhibited the proliferation,invasion,and migration of SKOV3 and ES-2 human ovarian cancer cells,inhibited the epithelial mesenchymal transformation of ovarian cancer,and promoted apoptosis of ovarian cancer cells.The mechanism may be related to the inhibition of the WNT/β-catenin signaling pathway.