着丝粒蛋白K调控YAP1对结直肠癌细胞增殖和侵袭迁移的影响  

Effects of cetromere protein K on proliferation,invasion and migration of colorectal cancer cells by regulating YAP1

作　　者：周琳 龚莲 孔丹丹 谢宇锋[2] 陶敏[3] ZHOU Lin;GONG Lian;KONG Dandan;XIE Yufeng;TAO Min(Department of Oncology, the First Affiliated Hospital of Soochow University, Suzhou 215006, China)

机构地区：[1]苏州大学附属太仓医院血液肿瘤科,215400 [2]苏州大学附属第一医院肿瘤科,江苏苏州215006 [3]苏州大学附属独墅湖医院肿瘤科,215124

出　　处：《临床肿瘤学杂志》2021年第12期1057-1064,共8页Chinese Clinical Oncology

基　　金：国家自然科学基金资助项目(81772645)。

摘　　要：目的探讨着丝粒蛋白K(CENPK)在结直肠癌组织中的表达及临床意义并探索CENPK与结直肠癌细胞生物学功能间的关系。方法采用实时定量PCR(qPCR)法检测43对结直肠癌组织和正常组织及结直肠癌细胞和人正常肠黏膜上皮细胞的CENPK水平,分析CENPK水平与结直肠癌临床病理特征的关系,选择CENPK低表达的结直肠癌细胞转染pcDNA3.1/CENPK质粒进行过表达CENPK处理,选择CENPK高表达的结直肠癌细胞转染si-CENPK进行敲减CENPK处理。采用CCK-8法、EdU实验、Transwell侵袭实验和划痕实验检测细胞增殖、DNA合成及侵袭迁移能力的变化情况,Western blotting检测Yes相关蛋白1(YAP1)和上皮间质转化(EMT)相关指标的蛋白水平。结果结直肠癌组织的CENPK水平高于正常组织(P<0.05),且其水平与肿瘤大小、脉管侵袭和淋巴结转移有关(P<0.05)。体外实验表明,干扰CENPK表达后,结直肠癌细胞的增殖、迁移和侵袭能力减弱;而过表达CENPK则会促进结直肠癌细胞的增殖、迁移和侵袭能力。干扰CENPK可下调YAP1水平而过表达CENPK可上调YAP1水平进而影响EMT相关蛋白的表达。YAP1过表达部分恢复CENPK沉默对细胞增殖、迁移、侵袭和EMT的抑制作用。结论CENPK在结直肠癌组织中异常高表达,可通过调控YAP1来促进人结直肠癌细胞的增殖、EMT、迁移和侵袭,CENPK/YAP1轴在结直肠癌防治中有一定价值。Objective To investigate the expression and clinical significance of cetromere protein K(CENPK)in colorectal cancer tissues,and to explore the relationship between CENPK and the biological function of colorectal cancer cells.Methods CENPK levels in 43 pairs of colorectal cancer tissues and adjacent normal tissues as well as colorectal cancer cells and human normal intestinal mucosal epithelial cells were detected by real-time quantitative PCR(qPCR).Relationship between CENPK expression and clinciopathological features of patients with colorectal cancer was studied.Colorectal cancer cells with low CENPK expression were transfected with pcDNA3.1/CENPK plasmid as to enhance CENPK expression,and colorectal cancer cells with high CENPK expression were transfected with si-CENPK as to silence CENPK expression.CCK-8,EdU assay,Transwell invasion experiment and scratch experiment were used to detect the changes in abilities of cell proliferation,DNA synthesis,invasion and migration.Protein levels of Yes-associated protein 1(YAP1)and epithelial mesenchymal transformation(EMT)related factors were detected by Western blotting.Results CENPK level in colorectal cancer tissues was higher than that in normal tissues(P<0.05),and its level was related to tumor size,vascular invasion and lymph node metastasis(P<0.05).In-vitro experiments showed that the proliferation,migration and invasion of colorectal cancer cells decreased after interfering CENPK expression,while overexpression of CENPK can promote the proliferation,migration and invasion of colorectal cancer cells.Interferencing CENPK can down-regulate the YAP1 level,while overexpression of CENPK can up-regulate the YAP1 level,thus affect the expression of EMT-related proteins.YAP1 overexpression partially restored the inhibitory effects of CENPK silencing on cell proliferation,migration,invasion and EMT.Conclusion The expression of CENPK was upregulated in colorectal cancer tissues,and it may promote colorectal cancer cells proliferation,invasion,migration and EMT in human colo

关 键 词：结直肠癌 着丝粒蛋白K Yes相关蛋白1 上皮间质转化 

分 类 号：R735.3[医药卫生—肿瘤]