| 引用本文: | 宋琳琳,孟焕然,周丽娜,刘瑞,殷利军*.LncRNA OIP5-AS1对卵巢上皮IOSE80表型转化的影响[J].中国现代应用药学,2024,41(5):649-656. |
| SONG Linlin,MENG Huanran,ZHOU Lina,LIU Rui,YIN Lijun*.Effect of LncRNA OIP5-AS1 on Phenotypic Transformation of IOSE80 in Ovarian Epithelium[J].Chin J Mod Appl Pharm(中国现代应用药学),2024,41(5):649-656. |
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| 摘要: |
| 目的 探索lncRNA OIP5-AS1对卵巢上皮细胞IOSE80发生增殖、迁移、凋亡、侵袭和周期的表型的变化及可能的机制。方法 患者临床资料数据收集自TCGA数据库及GEO数据库，R包分析后可视化OIP5-AS1在卵巢癌组织中表达上调，生存率与OIP5-AS1的相关性采用Kaplan-Meier分析。以慢病毒载体构建OIP5-AS1过表达和沉默的IOSE80细胞模型，采用RT-qPCR验证OIP5-AS1的表达，CCK-8检测细胞增殖，Transwell检测侵袭，划痕试验检测细胞迁移，流式细胞术检测细胞周期和凋亡，Western blotting检测细胞侵袭相关蛋白E钙黏着蛋白(E-cadherin)和N钙黏着蛋白(N-cadherin)的表达及细胞周期蛋白依赖性激酶(cyclin-dependent kinase，CDK)和细胞周期蛋白G相关激酶(cyclin-G-related kinase，GAK)的表达。结果 RT-qPCR结果显示成功构建OIP5-AS1过表达和沉默的IOSE80细胞株。CCK-8结果显示过表达OIP5-AS1促进IOSE80细胞的增殖。划痕试验结果显示过表达OIP5-AS1促进IOSE80细胞迁移。Transwell结果显示过表达OIP5-AS1会引起IOSE80细胞的侵袭力增强。流式细胞术结果表明OIP5-AS1的过表达使IOSE80细胞凋亡减弱并推动了细胞周期的进展。Western blotting结果显示过表达OIP5-AS1会下调E-cadherin的表达并上调N-cadherin的表达，同时过表达OIP5-AS1可提高CDK及GAK蛋白的表达。结论 lncRNA OIP5-AS1通过上调CDK及GAK的表达进一步干预了IOSE80细胞周期的调控，进而实现对卵巢上皮细胞恶性表型的间接调控作用。 |
| 关键词: 卵巢上皮细胞 lncRNA OIP5-AS1 细胞周期 表型 |
| DOI：10.13748/j.cnki.issn1007-7693.20224213 |
| 分类号:R966 |
| 基金项目:宁夏自然科学基金项目(2021AAC03332) |
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| Effect of LncRNA OIP5-AS1 on Phenotypic Transformation of IOSE80 in Ovarian Epithelium |
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SONG Linlin1, MENG Huanran1, ZHOU Lina1, LIU Rui1, YIN Lijun*2
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1.Department of Gynecology, The General Hospital of Ningxia Medical University, Yinchuan 750001, China;2.Department of General Surgery, People's Hospital of Ningxia Hui Autonomous Region, Yinchuan 750001, China
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| Abstract: |
| OBJECTIVE To explore the phenotypic changes and possible mechanisms of lncRNA OIP5-AS1 on the proliferation, migration, apoptosis, invasion and cycle of ovarian epithelial cells IOSE80. METHODS The clinical data of patients were collected from TCGA database and GEO database. After R package analysis, the differential expression of OIP5-AS1 was visualized in the volcanic map. The correlation between survival rate and OIP5-AS1 was analyzed by Kaplan-Meier. The IOSE80 cell model of OIP5-AS1 over expression and silencing was constructed with lentivirus vector. The expression of OIP5-AS1 was verified by RT-qPCR. Cell proliferation was detected by CCK-8. Invasion was detected by Transwell. Cell migration was detected by scratch test. Cell cycle and apoptosis were detected by flow cytometry. Western blotting was used to detect the expression of E-cadherin and N-cadherin, as well as the expression of cyclin-dependent kinase(CDK) and cyclin-G-related kinase(GAK). RESULTS RT-qPCR results showed that IOSE80 cell lines over expressing and silencing OIP5-AS1 were successfully constructed. CCK-8 results showed that overexpressing OIP5-AS1 promoted the proliferation of IOSE80 cells. Scratch test results showed that overexpressing OIP5-AS1 promoted the migration of IOSE80 cells. Transwell results showed that overexpressing OIP5-AS1 would increase the invasiveness of IOSE80 cells. Flow cytometry results showed that overexpression of OIP5-AS1 weakened the apoptosis of IOSE80 cells and promoted the progress of cell cycle. Western blotting results showed that overexpression of OIP5-AS1 downregulated the expression of E-cadherin and upregulated the expression of N-cadherin, while overexpression of OIP5-AS1 increased the expression of CDK and GAK proteins. CONCLUSION LncRNA OIP5-AS1 further interferes with the regulation of IOSE80 cell cycle by up regulating the expression of CDK and GAK, and then indirectly regulates the malignant phenotype of ovarian epithelial cells. |
| Key words: ovarian epithelial cell lncRNA OIP5-AS1 cell cycle phenotype |
