LncRNA ZEB2-AS1联合亲本基因ZEB2在卵巢癌SKOV3细胞中的作用及其机制研究OACSTPCD
Functional study of LncRNA ZEB2-AS1 and parental gene ZEB2 in promoting epithelial-mesenchymal transition in ovarian cancer SKOV3 cells
目的 探索LncRNAZEB2-AS1联合亲本基因ZEB2在卵巢癌SKOV3细胞中的功能及可能的机制.方法 原位杂交和免疫组织化学分别检测ZEB2-AS1和ZEB2在卵巢癌组织中的表达;建立ZEB2-AS1的过表达和沉默SKOV3细胞模型;CCK-8法检测细胞增殖能力;流式细胞术检测细胞凋亡和周期;细胞免疫荧光和Western blotting检测上皮-间充质相互作用(EMT)相关蛋白的表达;双萤光素酶基因报告实验检测ZEB2-AS1与ZEB2、microRNA-145(miR-145)的结合关系.结果 ZEB2-AS1和ZEB2均表达于卵巢癌组织中的细胞质和细胞核;相比SKOV3细胞,ZEB2-AS1过表达的SKOV3细胞增殖能力增强(P<0.05),凋亡能力减弱(P<0.05),侵袭和迁移能力增强(P<0.05),S期细胞数增加(P<0.05),G1期细胞数减少(P<0.05);EMT相关蛋白 E-cadherin 相对表达量降低(P<0.05),N-cadherin 和 Vimentin 相对表达量升高(P<0.05);而 ZEB2-AS1沉默的SKOV3细胞则相反.双萤光素酶报告实验结果证实,ZEB2-AS1和miR-145具有结合关系,ZEB2-AS1与ZEB2 mRNA具有结合关系.结论 LncRNA ZEB2-AS1联合亲本基因ZEB2在卵巢癌SKOV3细胞中发挥促EMT的功能作用.
Objective To explore the function and potential mechanism of LncRNA ZEB2-AS1 combined with its parental gene ZEB2 in ovarian cancer SKOV3 cells.Methods In situ hybridization and immunohistochemistry were used to detect the expression of ZEB2-AS1 and ZEB2 in ovarian cancer tissues.SKOV3 cell models with ZEB2-AS1 overexpression and silencing were established.Cell proliferation was measured using the CCK-8 assay.Flow cytometry was used to detect cell apoptosis and cycle.Epithelial-mesenchymal transition(EMT)-related protein expression was assessed through immunofluorescence and Western blotting.Dual-luciferase gene reporter assays were performed to confirm the binding relationship between ZEB2-AS1,ZEB2,and microRNA-145(miR-145).Results ZEB2-AS1 and ZEB2 were expressed in the cytoplasm and nucleus of ovarian cancer tissues.Compared with SKOV3 cells,ZEB2-AS1 overexpressing SKOV3 cells showed increased proliferation(P<0.05),reduced apoptosis(P<0.05),enhanced invasion and migration(P<0.05),increased S-phase cells(P<0.05),and decreased G,phase cells(P<0.05).EMT-related protein expression showed a decrease in E-cadherin(P<0.05)and an increase in N-cadherin and Vimentin(P<0.05).Conversely,ZEB2-AS1 silenced SKOV3 cells exhibited opposite trends.Dual-luciferase reporter assay confirmed the binding relationship between ZEB2-AS1 and miR-145,as well as between ZEB2-AS1 and ZEB2 mRNA.Conclusions LncRNA ZEB2-AS1 combined with its parental gene ZEB2 promotes EMT in ovarian cancer SKOV3 cells.
董辉;王晶;杨丹;丁永慧
宁夏医科大学总医院医学科学研究院,宁夏银川 750004宁夏医科大学总医院病理科,宁夏银川 750004宁夏医科大学第一临床医学院,宁夏银川 750004宁夏医科大学总医院妇科,宁夏 银川 750004
临床医学
卵巢癌LncRNA ZEB2-AS1ZEB2上皮-间充质相互作用
ovarian cancerLncRNA ZEB2-AS1ZEB2epithelial-mesenchymal transition
《中国现代医学杂志》 2024 (013)
28-40 / 13
宁夏自然科学基金(NO:2023AAC03601);宁夏回族自治区重点研发计划(NO:2019BEH03005)
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