CRABP2通过Wnt/β-catenin通路调控子宫内膜样腺癌细胞的增殖与侵袭

CRABP2 regulates the proliferation and invasion of endometrioid adenocarcinoma cells through the Wnt/β-catenin pathway

目的:探讨细胞视黄酸结合蛋白2(CRABP2)在子宫内膜样腺癌组织和细胞中的表达及其对子宫内膜样腺癌细胞增殖与侵袭的影响与其分子机制。方法:收集2020年6月至2021年4月在衡水市人民医院手术切除的子宫内膜样腺癌组织及配对正常子宫内膜组织,共24对;体外培养人子宫内膜样腺癌细胞An3ca、KLE。采用免疫组化分析及WB法检测子宫内膜样腺癌组织及正常子宫内膜组织中CRABP2的表达情况,采用WB法检测An3ca及KLE细胞中敲低CRABP2表达的效率,并以EDU法及Transwell实验检测敲低CRABP2表达后的An3ca及KLE细胞的增殖及侵袭能力,免疫荧光染色及WB法检测敲低CRABP2表达后的An3ca及KLE细胞中Wnt/β-catenin通路相关关键蛋白(β-catenin、c-Myc、cyclin-D1、MMP7及MMP9)的表达情况。以裸鼠体内成瘤实验观察敲低CRABP2表达对子宫内膜样腺癌细胞移植瘤生长和移植瘤组织中Ki67和β-catenin表达的影响。结果:与正常子宫内膜组织相比,CRABP2在人子宫内膜样腺癌组织中表达上调(P<0.01)。转染靶向CRABP2的shRNA后,An3ca、KLE细胞中CRABP2的表达降低(均P<0.01);敲低CRABP2表达后An3ca及KLE细胞增殖及侵袭能力均降低(均P<0.01),并且Wnt/β-catenin通路受到抑制(P<0.01)。成瘤实验显示敲低CRABP2表达后,裸鼠体内移植瘤体积明显缩小(P<0.01)。结论:CRABP2可通过调节Wnt/β-catenin通路发挥对子宫内膜样腺癌细胞增殖与侵袭的促进作用。

Objective:To investigate the expression of cellular retinoic acid binding protein 2(CRABP2)in endometrial adenocarcinoma tissues and cells and its effects on the proliferation and invasion of endometrial adenocarcinoma cells as well as its molecular mechanism.Methods:A total of 24 pairs of endometrioid adenocarcinoma tissues and matched normal endometrial tissues were collected from Hengshui People’s Hospital from June 2020 to April 2021,and human endometrial adenocarcinoma cells(An3ca and KLE)were cultured in vitro.The expression of CRABP2 in human endometrioid adenocarcinoma tissues and normal endometrial tissues was analyzed by immunohistochemistry and WB.The knockdown efficiency of CRABP2 in An3ca and KLE cells was detected by WB method,and the proliferation and invasion ability of An3ca and KLE cells after CRABP2 knockdown was detected by EDU method and Transwell assay,the expressions of key proteins related to Wnt/β-catenin pathway(β-catenin,c-Myc,cyclin-D1,MMP7and MMP9)in An3ca and KLE cells after CRABP2 knockdown were detected by WB.The effects of CRABP2 on the growth of xenograft tumor and the expression of Ki67 andβ-catenin in xenograft tumor tissues were observed in nude mice tumorigenesis assay.Results:Compared with normal endometrial tissue,the expression of CRABP2 was up-regulated in human endometrioid adenocarcinoma tissue.Expression of CRABP2 in An3ca and KLE cells decreased after transfection of shRNA targeting CRABP2(all P<0.01).The proliferation and invasion ability of An3ca and KLE cells were decreased after CRABP2 knockdown(all P<0.01)and the Wnt/β-catenin pathway was inhibited(P<0.01).The tumor formation experiment showed that the tumor volume of transplanted tumor in nude mice decreased significantly after CRABP2 knockdown(P<0.01).Conclusion:CRABP2 can promote the proliferation and invasion of endometrioid adenocarcinoma cells by regulating the Wnt/β-catenin pathway.