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

Functional study of LncRNA ZEB2-AS1 and parental gene ZEB2 in promoting epithelial-mesenchymal transition in ovarian cancer SKOV3 cells

摘要:

目的 探索LncRNA ZEB2-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。

Abstract:

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 G₁ 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.