BAP18对卵巢颗粒细胞瘤基因调控和细胞迁移的影响

Effect of BAP18 on gene regulation and cell migration in granulosa cell tumor of ovary

目的研究沉默BAP18对重要促癌基因在人卵巢颗粒细胞瘤细胞中表达和细胞迁移能力的影响。方法利用免疫荧光的方法研究BAP18蛋白在卵巢颗粒细胞瘤细胞中的表达及定位。用慢病毒感染KGN和COV434细胞,构建稳定敲低BAP18基因的两种卵巢颗粒细胞细胞系,应用实时定量PCR方法检测细胞中AMH、Notch2、Notch3的mRNA表达,并通过细胞划痕实验,对比沉默BAP18组和对照组细胞的迁移能力。结果BAP18在两种卵巢颗粒细胞瘤细胞中均有表达,主要定位于细胞核。与对照组相比,沉默BAP18组的细胞中AMH、Notch2 mRNA表达水平显著降低,但Notch3 mRNA表达并无影响,且沉默BAP18组的细胞迁移能力显著减弱。结论沉默BAP18基因表达减弱卵巢颗粒细胞瘤细胞迁移能力与抑制促癌基因Notch2和AMH的表达相关。

Objective To study the effect of knockdown BAP18 on important oncogene expression and cell migration ability in granulosa cell tumor.Methods The expression and localization of BAP18 protein in granulosa cells were detected by immunofluorescence.KGN and COV434 cells were infected with lentivirus to construct a stable granulosa cell line that knocked down BAP18 genes.RT quantitative PCR was used to detect the expression of AMH,Notch2 and Notch3 mRNAs.Cell wound scratch was used to determine the migration ability of the cells.Results BAP18 was expressed positively in KGN and COV434 cells,and mainly located in the nucleus.The expression level of AMH and Notch2 mRNAs was significantly decreased,but with not affection for expression of Notch3 mRNA,and the cell migration ability was significantly decreased in the silent BAP18 group than in control group.Conclusion The decreased expression of BAP18 gene inhibite migration ability of granulosa cells,which is related to the expression of oncogene Notch2 and AMH.