文章摘要

整合素连接激酶对鼻咽癌细胞系迁移及上皮- 间充质转化的影响

作者: 1吴平安, 2赵丹芸, 1梁秀妮, 3唐青来, 3李蒙蒙, 3李仕晟, 1朱洪蕾, 1曾敬贤
1 香港大学深圳医院耳鼻咽喉头颈外科，广东 深圳 518053
2 香港大学深圳医院中心实验室，广东 深圳 518053
3 中南大学湘雅二医院耳鼻咽喉头颈外科，长沙 410011
通讯: 吴平安 Email: wupa@hku-szh.org
DOI: 10.3978/j.issn.2095-6959.2019.06.002
基金: 深圳市科技计划项目(JCYJ20160429190356200)；深圳市卫生与计划生育系统科研项目(201607027)。

摘要

目的：探讨整合素连接激酶(integr in-linked k inase，ILK)对鼻咽癌细胞迁移及上皮-间充质转化(epithelial-mesenchymal transition，EMT)转化的影响及其作用机制。方法：通过脂质体介导将ILK SiRNA转染CNE2鼻咽癌细胞系，细胞分为siILK组、siGAPDH阳性对照组及siControl阴性对照组。 RT-PCR和Western印迹检测鼻咽癌细胞系CNE2中ILK的RNA水平和蛋白水平表达的变化。Transwell实验和划痕试验观察ILK SiRNA对细胞迁移能力及侵袭能力的影响，RT-PCR法检测了与EMT过程中的相关基因mRNA的表达水平。结果：转染ILK SiRNA后，CNE2细胞中ILK mRNA和蛋白表达水平均明显下降；ILK SiRNA可明显抑制CNE2细胞的迁移及侵袭能力；上调E-cadherin的表达，下调OCLN，Vimentin，Twist1，FN1的表达，抑制EMT过程的发生。结论：ILK在鼻咽癌细胞系CNE2中高表达，降低ILK表达可诱导鼻咽癌细胞间充质-上皮细胞转型的发生，并因此而显著降低其侵袭和转移潜力。
关键词： 整合素连接激酶；鼻咽癌；生物标志物；转移；细胞迁移；上皮-间充质转化

Effects of ILK regulating epithelial-mesenchymal transition on invasion and migration in nasopharyngeal cancer

Authors: 1WU Ping’an, 2ZHAO Danyun, 1LIANG Xiuni, 3TANG Qinglai, 3LI Mengmeng, 3LI Shisheng, 1ZHU Honglei, 1Raymond K. Tsang
1 Department of Otolaryngology, Head and Neck Surgery, University of Hong Kong-Shenzhen Hospital, Shenzhen Guangdong 518053, China
2 Central Laboratory, University of Hong Kong-Shenzhen Hospital, Shenzhen Guangdong 518053, China
3 Department of Otolaryngology-Head and Neck Surgery, Second Xiangya Hospital, Central South University, Changsha 410011, China

CorrespondingAuthor: WU Ping’an Email: wupa@hku-szh.org

DOI: 10.3978/j.issn.2095-6959.2019.06.002

Foundation: This work was supported by the Shenzhen Science and Technology Plan Project (JCYJ20160429190356200) and Shenzhen Health and Family Planning System Research Project (201607027), China.

Abstract

Objective: To investigate the effect of integrin-linked kinase (ILK) on migration and epithelial-mesenchymal transition of nasopharyngeal carcinoma cells and its mechanism. Methods: ILK SiRNA was transfected into CNE2 nasopharyngeal carcinoma cell line by liposome. The cells were divided into siILK group, positive control group siGAPDH and siControl negative control group. RT-PCR and Western blot were used to detect the changes of ILK RNA and protein levels in nasopharyngeal carcinoma cell line CNE2. The effects of ILK siRNA on cell migration and invasion were observed by Transwell assay and scratch assay. The expression level of related Marker in epithelial-mesenchymal transition (EMT) was detected by RT-PCR. Results: After transfection of ILK siRNA, the expression of ILK mRNA and protein in CNE2 cells decreased significantly. ILK siRNA could significantly inhibit the migration and invasion of CNE2 cells; up-regulate the expression of E-cadherin and down-regulate the expression of OCLN, Vimentin, Twist1 and FN1, inhibit the occurrence of epithelial-mesenchymal transition process. Conclusion: ILK is highly expressed in the nasopharyngeal carcinoma cell line CNE2. Decreasing ILK expression induces the development of mesenchymal-epithelial cell transition in nasopharyngeal carcinoma cells, and thus significantly reduces its invasion and metastatic potential.
Keywords： integrin-linked kinase; nasopharyngeal carcinoma; biomarkers; metastasis; cell migration; epithelial-mesenchymal transition

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