摘要
目的 研究长链非编码RNA MALAT1(lncRNA MALAT1)在卵巢癌中的作用及调控机制。方法 实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction, RT-qPCR)检测MALAT1在人正常卵巢上皮细胞系IOSE80及人卵巢癌细胞系SKOV3、OVCA429和HO-8910PM中的表达,选取SKOV3及HO-8910PM细胞进行后续研究。利用siRNA干扰SKOV3和HO-8910PM细胞中MALAT1表达,CCK-8法检测细胞增殖,划痕及Transwell小室检测细胞迁移及侵袭能力,并通过Western blot法检测上皮-间充质转化(epithelial-mesenchymal transformation, EMT)相关指标E-cadherin、N-cadherin的表达。利用生物信息学分析MALAT1与miRNA-141-3p的靶向关系,并利用双荧光素报告基因验证。MALAT1敲低及miRNA-141-3p抑制剂共同作用细胞,检测其对SKOV3及HO-8910PM细胞增殖、侵袭及迁移的影响,并通过Western blot法分析其对Wnt/β-catenin信号通路相关蛋白的表达调控。结果 与人正常卵巢上皮细胞系IOSE80比较,卵巢癌细胞系内MALAT1表达明显上调(P<0.05);而在SKOV3及HO-8910PM中下调MALAT1表达,细胞增殖、侵袭迁移及EMT均受到抑制,同时miRNA-141-3p表达上调(P<0.05)。双荧光素酶报告基因结果证明MALAT1和miRNA-141-3p具有靶向关系。而在下调MALAT1表达的同时使用miRNA-141-3p抑制剂,则可逆转下调MALAT1的所产生的抑制效应(P<0.05)。进一步的研究发现,MALAT1/miRNA-141-3p轴可能通过调控Wnt/β-catenin信号通路影响卵巢癌进展。结论 MALAT1可能通过海绵吸附miRNA-141-3从而调控Wnt/β-catenin影响卵巢癌的发生和发展。
Objective To study the role and regulatory mechanism of long non-coding RNA MALAT1 in ovarian cancer.Methods Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of MALAT1 in human normal ovarian epithelial cell line IOSE80 and human ovarian cancer cell line SKOV3,OVCA429 and HO-8910PM.SKOV3 and HO-8910PM cells were selected for the next study.The expression of MALAT1 in SKOV3 and HO-8910PM cells were interfered with siRNA,the cell proliferation was detectd by CCK-8 method,the cell migration and invasion were detectd by scratche and Transwell assay respectively,and the expression of epithelial-mesenchymal transformation(EMT)-related indicators E-cadherin and N-cadherin were detected by Western blot.The targeting relationship between MALAT1 and miRNA-141-3p was analyzed by bioinformatics,and was verified by dual luciferase reporter.MALAT1 knockdown and miRNA-141-3p inhibitor were treated the SKOV3 and HO-8910PM cells at the same time,then the cell proliferation,invasion and migration were detected,Western blot was used to detect the expression of Wnt/β-catenin signaling pathway related proteins.Results Compared with human normal ovarian epithelial cell line IOSE80,the expression of MALAT1 in ovarian cancer cell line was significantly up-regulated(P<0.05).When the expression of MALAT1 in SKOV3 and HO-8910PM was down-regulated,the cell proliferation,invasion,migration and EMT were inhibited(P<0.05).At the same time,the expression of miRNA-141-3p was up-regulated(P<0.05),and the results of dual luciferase reporter proved that there was a targeting relationship between MALAT1 and miRNA-141-3p.However,the inhibitory effect of MALAT1down-regulation in the cells can be reversed by miRNA-141-3p inhibitor(P<0.05).Further studies had found that the MALAT1/miRNA-141-3p axis may influence the progression of ovarian cancer by regulating the Wnt/β-catenin signaling pathway.Conclusion MALAT1may influence the occurrence and development of ovarian cancer by sponge adsorption of miRNA-141-3 via regulating Wnt/β-catenin.
作者
金烨
JIN Ye(Department of Oncology,Women′s Hospital,School of Medicine,Zhejiang University,Zhejiang 310006,China)
出处
《医学研究杂志》
2024年第4期115-121,共7页
Journal of Medical Research
基金
浙江省基础公益研究计划项目(LQ20H040009)。