摘要
结直肠癌(colorectal cancer,CRC)是一种危害人类健康的恶性肿瘤。跨膜糖蛋白Embigin(EMB)是免疫球蛋白超家族成员之一,参与调控肿瘤细胞的增殖和迁移,然而,其在结直肠癌进展中的作用和机制仍不清楚。本研究旨在分析EMB在结直肠癌中的表达及其临床意义,研究其对结直肠癌细胞增殖的影响及其作用机制。免疫组织化学、实时荧光定量PCR和蛋白质印迹法结果表明,EMB在结直肠癌组织和细胞中表达下调(P<0.01),EMB的低表达与结直肠癌的不良分化、TNM分期和患者的不良预后呈显著正相关关系(P<0.05)。CCK-8细胞增殖实验、平板克隆形成实验和裸鼠皮下成瘤实验结果表明,抑制EMB可促进结直肠癌细胞的增殖(P<0.01)。RNA-seq和生物信息学分析的结果表明,EMB参与了细胞周期进程。流式细胞术、蛋白质印迹法和回复实验结果表明,干扰EMB可促进结直肠癌细胞细胞周期G_(1)/S期转变(P<0.05)。干扰EMB对细胞增殖的影响可被CDK4/6抑制剂Palbociclib(PD-0332991)HCl逆转(P<0.001)。综上所述,EMB低表达能够通过促进细胞周期G_(1)/S期转变促进结直肠癌细胞的增殖。本研究可能为结直肠癌的治疗提供一个新的靶点。
Colorectal cancer(CRC)is a common malignancy that is harmful to people worldwide.The transmembrane glycoprotein embigin(EMB)is a member of the immunoglobulin superfamily which has been demonstrated to be involved in the proliferation and metastasis of tumor cell.However,the role and mechanism of EMB in the progression of CRC remain unclear.This study analyzes the expression and clinical significance of EMB in CRC,investigates its effect on the proliferation of CRC cells and explores its mechanism.The results of immunohistochemistry,real-time quantitative PCR(RT-qPCR)and Western blotting showed that the expression of EMB was down-regulated in CRC tissues and cells(P<0.01),and the down-regulation of EMB was positively correlated with poor differentiation,TNM staging and poor prognosis of the patients(P<0.05).The results of the CCK-8 cell proliferation assay,colony formation assay and nude mice tumorigenecity assay showed that inhibition of EMB could promote the proliferation of CRC cells(P<0.01).RNA-seq and bioinformatics analysis showed that EMB participated in the cell cycle process.The results of flow cytometry,Western blotting and rescue experiments showed that EMB knockdown could promote G_(1)/S cell cycle transition in CRC cells(P<0.05)and the effect of EMB knockdown on CRC cell proliferation could be reversed by CDK4/6 inhibitor Palbociclib(PD-0332991)HCl(P<0.05).In summary,down-regulation of EMB could promote the proliferation of CRC cells by facilitating G_(1)/S transition,and this study might provide a new therapeutic target for CRC patients.
作者
胡育菡
范文艳
马帅
钟加滕
王海军
HU Yu-Han;FAN Wen-Yan;MA Shuai;ZHONG Jia-Teng;WANG Hai-Jun(Department of Pathology,School of Basic Medical Sciences,Xinxiang Medical University,Xinxiang 453003,Henan,China;Micromorphology Laboratory,School of Basic Medical Sciences,Xinxiang Medical University,Xinxiang 453003,Henan,China;Department of Pathology,The First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,Henan,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2021年第12期1611-1620,共10页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金项目(No.U1904133)
河南省自然科学基金项目(No.212300410223)
河南省科技攻关项目(No.202102310091)
新乡医学院博士科研启动基金项目(No.XYBSKYZZ201820)
新乡医学院教学改革研究项目(No.2019-XYJG-24)资助
河南省高等学校青年骨干教师培养计划项目(No.2020GGJS150)。
作者简介
通讯作者:王海军,Tel:0373-3029123,E-mail:wnavy200299@163.com。
