Calreticulin通过诱导细胞EMT促进鼻咽癌迁移和侵袭  

Calreticulin promotes migration and invasion of nasopharyngeal carcinoma cells by inducing cell EMT

作　　者：朱军辉[1] 金明[1] 邱浩[1] 张和良[1] 马为 肖玄 李承松 罗招阳[1] 张志伟[1] ZHU Jun-hui;JIN Ming;QIU Hao;ZHANG He-liang;MA Wei;XIAO Xuan;LI Chert -song;LUO Zhao-yang;ZHANG Zhi-wei(Cancer Institute of Medical College, Key Laboratory of Cancer Cellular and Molecular Pathology in Hunan Province;Department of Clinical Medicine of Medical College, University of Southern China, Hengyang 421001, China)

机构地区：[1]南华大学医学院肿瘤研究所,肿瘤细胞与分子病理学湖南省重点实验室,湖南衡阳421001 [2]南华大学医学院临床医学系,湖南衡阳421001

出　　处：《中国病理生理杂志》2018年第5期925-929,共5页Chinese Journal of Pathophysiology

基　　金：湖南省高校创新平台基金资助项目(No.12 K094;No.13 K083); 湖南省教育厅课题(No.11 C1112); 湖南省卫生厅课题(No.B2013-048;No.2014-163); 湖南省研究生科研创新项目(No.CX2016B478); 南华大学博士科研启动基金(No.2016 XQD21)

摘　　要：目的:观察钙网蛋白(calreticulin,CRT)在鼻咽癌组织中的表达并分析其临床病理学意义,揭示其对鼻咽癌CNE2细胞上皮-间充质转化(EMT)的影响。方法:S-P免疫组化法检测CRT在52例鼻咽癌组织和57例鼻咽部良性病变组织中的表达情况,并分析其临床病理学意义;构建特异性干扰CRT基因的小干扰RNA干扰载体,瞬时转染鼻咽癌CNE2细胞,观察CRT低表达对CNE2细胞形态的影响;Transwell迁移和侵袭实验检测CRT低表达对CNE2细胞迁移和侵袭能力的影响;Western blot检测CRT低表达对上皮型钙黏蛋白(E-cadherin)、波形蛋白(vimentin)、转化生长因子-β(TGF-β)及基质金属蛋白酶9(MMP-9)等EMT相关蛋白表达的影响。结果:CRT在鼻咽癌组织中的阳性表达率为82.69%(43/52),在鼻咽部良性病变组织的阳性表达率为19.29%(11/57),CRT在鼻咽癌组织中表达显著增高(P<0.05);CRT的阳性表达率与鼻咽癌分期及淋巴结转移呈正相关(P<0.05)。敲降CRT呈低表达后,CNE2细胞由梭形演变为扁平和鹅卵石样,且细胞排列紧密,细胞的迁移侵袭能力减弱(P<0.05)。敲降CRT呈低表达后,E-cadherin表达显著增高,vimentin、TGF-β以及MMP-9蛋白的表达降低(P<0.05)。结论:Calreticulin在鼻咽癌组织中表达显著增高,且与鼻咽癌临床分期及淋巴结转移呈正相关。Calreticulin可诱导鼻咽癌CNE2细胞发生EMT,进而促进鼻咽癌的迁移和侵袭。AIM： To observe the expression of calreticulin（ CRT） in nasopharyngeal carcinoma tissues,analyze the significance of clinical pathology and the influence on epithelial-mesencymal transition（ EMT） of CNE2 cells.METHODS： The expression of calreticulin was detected by immunohistochemistry in 52 nasopharyngeal carcinoma and 57 nasopharyngeal benign tissues,and the significance of clinical pathology was evaluated. The calreticulin gene-specific small interfering RNA was constructed,and then was transfected into the NPC cell line CNE2 using the cationic liposome method. The effect of CRT on the morphological changes of the CNE2 cells was observed under light microscope. The effect of CRT on the cell migration and invasion abilities of the CNE2 cells was detected by Transwell migration and invasion assays.The expression of EMT-related proteins E-cadherin,vimentin,transforming growth factor（ TGF）-β and matrix metalloproteinase（ MMP）-9 in the CNE2 cells was determined by Western blot. RESULTS： The positive expression rate of CRT in the benign lesion tissues was 19. 29 %（ 11/57）,which was significantly increased in the nasopharyngeal carcinoma tissues as 82. 69 %（ 43/52）. The expression rate of CRT was positively correlated with the stage of nasopharyngeal carcinoma and lymph node metastasis（ P〈0. 05）. Knockdown of CRT expression made the CNE2 cells showing a spindle shape to a flat,cobblestone-like epithelial state change,arranged more compact,and the migration and invasion abilities were significantly decreased（ P〈0. 05）. Knockdown of CRT expression resulted in significant increase in the protein expression of E-cadherin,and the decreases in the protein expression of vimentin,TGF-β and MMP-9 in the CNE2 cells（ P〈0. 05）. CONCLUSION： Calreticulin expression in nasopharyngeal carcinoma is significantly higher and positively correlated with nasopharyngeal carcinoma stage and lymph node metastasis. Calreticulin promotes cell migration and invasion of nasopharyngeal carcinoma

关 键 词：鼻咽癌 钙网蛋白 上皮-间充质转化 细胞迁移 细胞侵袭 

分 类 号：R739.63[医药卫生—肿瘤] R730.23[医药卫生—临床医学]