LncRNA AC130710通过miR-129-5P/WNT4轴促进子宫内膜癌细胞增殖和上皮间质转化  

LncRNA AC130710 promotes endometrial cancer cell proliferation and epithelial mesenchymal transformation through miR-129-5p/WNT4 axis

作　　者：梁芳 刘广申 徐艳 Liang Fang;Liu Guangshen;Xu Yan(Changsha Maternal and Child Health Care Hospital Obstetrics and Gynecology,Changsha 410000,China;Department of Gynecology,the Fourth Hospital of Changsha,Changsha 410000,China)

机构地区：[1]湖南省长沙市第四医院妇产科,长沙410000 [2]湖南省长沙市妇幼保健院妇产科,长沙410000

出　　处：《中华细胞与干细胞杂志（电子版）》2022年第4期206-214,共9页Chinese Journal of Cell and Stem Cell（Electronic Edition）

摘　　要：目的探讨LncRNAAC130710通过miR-129-5P/WNT4轴对子宫内膜癌细胞(HEC-1A细胞)增殖、凋亡及上皮间质转化(EMT)的影响及机制研究。方法通过实时荧光定量PCR检测LncRNAAC130710、miR-129-5P和WNT4在子宫内膜癌细胞(HEC-1A细胞)和人子宫内膜上皮细胞(HEEC)中的表达。细胞分别转染(1)siRNANC、AC130710siRNA、WNT4 siRNA;(2))inhibitor NC、miR-129-5P inhibitor;(3)pcDNA-3.1(+)+mimics NC、pcDNAAC130710+mimicsNC、pcDNA-3.1(+)+miR-129-5Pmimics、pcDNA-AC130710+miR-129-5P mimics。MTT实验检测LncRNAAC130710、miR-129-5P和WNT4的表达对HEC-1A细胞增殖能力的影响;Westernblot检测LncRNAAC130710、miR-129-5P和WNT4的表达对HEC-1A细胞凋亡相关蛋白B淋巴细胞瘤-2基因相关蛋白X(Bax)、剪切的半胱氨酰天冬氨酸特异性蛋白酶-3(cleavedcaspase-3)、cleaved caspase-9和B淋巴细胞瘤-2基因(Bcl-2)表达的影响;Westernblot检测LncRNAAC130710、miR-129-5P和WNT4的表达对HEC-1A细胞EMT的影响。miRanda和双荧光素酶报告基因实验分析LncRNAAC130710和miR-129-5P之间的关系,TargetScan数据库分析miR-129-5P与WNT4的相关性,双荧光素酶报告基因检测miR-129-5P与WNT4的相互作用;RT-qPCR法检测LncRNAAC130710通过miR-129-5P对WNT4表达的影响。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析,两两比较采用LSD-t检验。结果与HEEC细胞比较,HEC-1A细胞中AC130710表达水平(1.86土0.21比0.85±0.06)、WNT4表达水平(1.88土0.26比1.08±0.12)升高;HEC-1A细胞中miR-129-5P表达水平(0.89土0.16比1.76土0.08)降低。与转染siRNANC比较,转染AC130710siRNA细胞内Bax、cleaved caspase-3、cleaved caspase-9、E-cadherin蛋白相对表达水平[(1.37±0.14比0.84±0.21),(1.08±0.16比0.37±0.07),(1.26±0.24比0.39±0.06),(1.87±0.17比1.32±0.26))上升,Bcl-2、N-cadherin、Snail和Vimentin蛋白相对表达水平[(0.38±0.08比1.18±0.14),(0.36±0.04比0.85±0.24),(0.35±0.09比1.12±0.18),(0.42±0.10比1.26±0.27)]下降;与转染inhibitorNCObjective To investigate the effect of LncRNA AC130710 on proliferation,apoptosis and epithelial mesenchymal transformation(EMT)of endometrial cancer cells(HEC-1A cells)through miR-129-5p/WNT4 axis and its mechanism.MethodsThe expressions of LncRNA AC130710,miR-129-5PandWNT4inHEC-1AandHEEC cellsweredetectedbyreal-time quantitative PCR(RT-qPCR),then the cells were transfected with(1)siRNANC,AC130710 siRNA and WNT4 siRNA;(2)inhibitor NC,miR-129-5p inhibitor;(3)pcDNA-3.1(+)+mimics NC,pcDNA-AC130710+mimics NC,pcDNA-3.1(+)+miR-129-5pmimics,pcDNA-AC130710+miR-129-5p mimics.The effects of LncRNA AC130710,miR-129-5P and WNT4 expressions on the proliferation ability of HEC-1A cells were detected by MTT experiment.Western blot was used to detect the effects of LncRNA AC130710,miR-129-5p and WNT4 on the expression of apoptosis-related proteins Bax,cleaved-caspase-3,cleaved-caspase-9 and Bcl-2 in HEC-1A cells.Western blot assay was used to detect the effects of LncRNA AC130710,miR-129-5P and WNT4 expressions on EMT of HEC-1A cells.The targeted relationship between LncRNA AC130710 and miR-129-5P was analyzed by miRanda and double luciferase reporter gene assay.The correlation between miR-129-5P and WNT4 was analyzed by the TargetScan database,and the interaction between miR-129-5P and WNT4 was detected by the double luciferase reporter gene.RT-qPCR detected the effect of LncRNA AC130710 on WNT4 expression through miR-129-5P.ResultsCompared with HEEC cells,AC130710 expression was increased in HEC-1A cells(1.86±0.21 vs 0.85±0.06).The expression level of miR-129-5p in HEC-1A cells was decreased(0.89±0.16 vs 1.76±0.08);Compared with HEEC cells,the expression of WNT4 in HEC-1A cells was increased(1.88±0.26 vs 1.08±0.12).Compared with the siRNA NC transfection group,the relative expression levels of Bax(1.37±0.14 vs 0.84±0.21),cleaved caspase-3(1.08±0.16vs 0.37±0.07),cleaved caspase-9(1.26±0.24vs 0.39±0.06),E-cadherin(1.87±0.17 vs 1.32±0.26)in AC130710 siRNA transfection group increased,the relative expression levels

关 键 词：Lnc RNA miR-129-5P WNT4 HEC-1A 细胞增殖 EMT 

分 类 号：R737.33[医药卫生—肿瘤]