FAM83H-AS1促进子宫内膜癌细胞系Ishikawa上皮-间质转化和血管生成OA
FAM83H-AS1 promotes epithelial-mesenchymal transition and angiogenesis in the endometrial cancer cell line Ishikawa
目的 探究FAM83H-AS1 对子宫内膜癌细胞上皮间质转化及血管生成的作用及机制.方法 人子宫内膜癌细胞Ishikawa培养后,分为空白组、FAM83H-ASsiRNA组(转染 FAM83H-ASsiRNA)、miR-4684-5p mimics组(转染 miR-4684-5p mimics)、FAM83H-AS siRNA+miR-4684-5p mimics 组(共转染 FAM83H-ASsiRNA 与miR-4684-5p mimics).Transwell小室法检测细胞的侵袭;Matrigel基质胶法检测细胞管腔形成;Western blot检测上皮间质转化标志蛋白E-cadherin和vimentin,双荧光素酶报告实验检测 FAM83H-AS1 与 miR-4684-5p靶向关系;qPCR检测FAM83H-AS1 及 miR-4684-5p表达.结果 与空白组比较,FAM83H-ASsiRNA组及 miR-4684-5p mimics组 Ishikawa 细胞中 FAM83H-AS1 降低、miR-4684-5p 升高(P<0.05);与 FAM83H-ASsiRNA 组及miR-4684-5p mimics组比较,FAM83H-AS siRNA+miR-4684-5p mimics 组 Ishikawa 细胞中 FAM83H-AS1 降低、miR-4684-5p升高(P<0.05),其中FAM83H-ASsiRNA组与miR-4684-5p mimics组的差异无统计学意义.与空白组比较,FAM83H-AS siRNA组及 miR-4684-5p mimics组细胞侵袭数量、管腔形成数量及蛋白 vimentin均降低,E-cadherin升高(P<0.05);与 FAM83H-ASsiRNA 组及 miR-4684-5p mimics 组比较,FAM83H-ASsiRNA+miR-4684-5p mimics组细胞侵袭数量、管腔形成数量及蛋白 vimentin 均降低,E-cadherin 升高(P<0.05),其中FAM83H-AS siRNA组与miR-4684-5p mimics 组的差异无统计学意义.双荧光素酶报告实验结果显示,转染miR-4684-5p mimics,FAM83H-AS1 3′UTR-WT组活性显著降低(P<0.05),但FAM83H-AS1 3′UTR-MUT组的影响并不显著.结论 在子宫内膜癌细胞系Ishikawa中,敲低FAM83H-AS1 及上调miR-4684-5p,抑制上皮-间质转化和血管生成,可能通过靶向调节miR-4684-5p发挥作用.
Objective To investigate the effect and mechanism of FAM83H-AS1 on epithelial-mesenchymal transi-tion(EMT)and angiogenesis in endometrial cancer cells.Methods Human endometrial cancer Ishikawa cells were divided into four groups:Blank group,FAM83H-AS1 siRNA group(transfected with FAM83H-AS1 siRNA),miR-4684-5p mimics group(transfected with miR-4684-5p mimics),and FAM83H-AS1 siRNA+miR-4684-5p mimics group(co-transfected with FAM83H-AS1 siRNA and miR-4684-5p mimics).Cell invasion was detected by Transwell assay;tube formation was detected by Matrigel assay;the EMT marker proteins E-cadherin and Vimentin were detected by Western blot;the targeted relationship between FAM83H-AS1 and miR-4684-5p was detected by dual-luciferase reporter assay;and the expression levels of FAM83H-AS1 and miR-4684-5p were detected by qPCR.Results Compared with the Blank group,the expression of FAM83H-AS1 was decreased and the expression of miR-4684-5p was increased in FAM83H-AS1 siRNA group and the miR-4684-5p mimics group(P<0.05).Compared with the FAM83H-AS1 siRNA group or the miR-4684-5p mimics group,the expression of FAM83H-AS1 was decreased and the expression of miR-4684-5p was increased in the FAM83H-AS1 siRNA+miR-4684-5p mimics group(P<0.05),among which there was no statistically significant difference between the FAM83H-AS1 siRNA group and the miR-4684-5p mimics group.Compared with the blank group,the number of cell invasions,tube formations,and the protein level of Vimentin were all decreased,while E-cadherin was in-creased in the FAM83H-AS1 siRNA group and the miR-4684-5p mimics group(P<0.05).Compared with the FAM83H-AS1 siRNA group or the miR-4684-5p mimics group,the number of cell invasions,tube formations,and the protein level of vimentin were all decreased,while E-cadherin was increased in the FAM83H-AS1 siRNA+miR-4684-5p mimics group(P<0.05),among which there was no statistically significant difference between the FAM83H-AS1 siRNA group and the miR-4684-5p mimics group.The dual-luciferase reporter assay showed that after transfection with miR-4684-5p mimics,the activity in the FAM83H-AS1 3′UTR-WT group was significantly reduced(P<0.05),whereas the effect on theFAM83H-AS1-3′UTR-MUTgroup was not significant.Conclusions In the endometrial cancer cell line Ishikawa,knockdown of FAM83H-AS1 or overexpression of miR-4684-5p inhib-its epithelial-mesenchymal transition and angiogenesis,an effect mediated by the targeted regulation of miR-4684-5p.
严鲁豫;王超;封硕
山东第二医科大学临床医学院 山东第二医科大学附属医院 妇科,山东 潍坊 261053山东第二医科大学临床医学院 山东第二医科大学附属医院 妇科,山东 潍坊 261053山东第二医科大学临床医学院 山东第二医科大学附属医院 妇科,山东 潍坊 261053
医药卫生
子宫内膜癌FAM83H-AS1miR-4684-5p上皮-间质转化血管生成
FAM83H-AS1miR-4684-5pendometrial cancerepithelial-mesenchymal transition(EMT)angiogenesis
《基础医学与临床》 2026 (4)
517-523,7
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