摘要
目的探讨miR-135b-5p对鼻咽癌细胞增殖、侵袭、转移的影响及其可能的机制。方法使用Lipofectamine-2000将miR-135b-5p siRNA转染至SUNE-1细胞,并将NC siRNA转染至SUNE-1细胞中作为对照组。实时定量PCR检测NP69、CNE-1、SUNE-1、C666-1细胞系及鼻咽癌组织中miR-135b-5p及TIMP3的表达水平。双荧光素酶报告基因检测miR-135b-5p及TIMP3的调控关系,免疫蛋白印迹实验检测TIMP3蛋白的表达含量,CCK-8实验检验SUNE-1细胞的增殖能力,Transwell实验检验SUNE-1细胞的迁移及侵袭能力。miR-135b-5p的表达与鼻咽癌(NPC)患者的临床病理学参数之间的关联。结果 miR-135b-5p在CNE-1、SUNE-1、C666-1细胞系及鼻咽癌组织中表达含量均上升,并且在SUNE-1细胞系中其的表达水平最低;双荧光素酶报告基因结果显示:抑制miR-135b-5p表达可显著增高SUNE-1细胞中TIMP3的荧光素酶活性;抑制miR-135b-5p表达后,TIMP3蛋白的表达相应上调,同时削弱了SUNE-1细胞增殖、迁移、侵袭能力。同时收集整理NPC患者相关临床资料进行对比分析,结果发现miR-135b-5p的表达与NPC的病理分期相关以及淋巴结转移情况呈正相关。结论抑制miR-135b-5p的表达可抑制鼻咽癌细胞的增殖、迁移和侵袭,并靶向上调TIMP3的表达。
Objective To investigate the effect of miR-135b-5p on proliferation, invasion and metastasis of nasopharyngeal carcinoma cells and its possible mechanism. Methods Real-time quantitative PCR was used to detect the expression levels of miR-135b-5p and TIMP3 in NP69, CNE-1, SUNE-1, C666-1 cell lines and nasopharyngeal carcinoma tissues. The dual luciferase reporter gene was used to detect the regulatory relationship between miR-135b-5p and TIMP3. Western blotting assay was used to detect the expression of TIMP3 protein, and CCK-8 assay was used to test the proliferation of SUNE-1 cells. Transwell assay was used to test the migration and invasion ability of SUNE-1 cells. Resluts The expression levels of miR-135b-5p in CNE-1, SUNE-1, C666-1 cell lines and nasopharyngeal carcinoma tissues increased, and the expression level was the lowest in SUNE-1 cell line. Double luciferase reporter gene results showed that inhibition of miR-135b-5p expression significantly increased TIMP3 luciferase activity in SUNE-1 cells. After inhibiting the expression of miR-135b-5p, the expression of TIMP3 protein was up-regulated, and the proliferation, migration and invasion of SUNE-1 cells were decreased. At the same time, we collected and analyzed the clinical data of NPC patients for comparative analysis. The results showed that the expression of miR-135b-5p was positively correlated with the pathological stage of NPC and lymph node metastasis. Conclusion Inhibition of miR-135b-5p expression can inhibit the proliferation, migration and invasion of nasopharyngeal carcinoma cells and up-regulate the expression of TIMP3.
作者
赵辉
董琼娜
金晓杰
ZHAO Hui; DONG Qiong-na; JIN Xiao-jie(Department of Otorhinolaryngology Head and Neck Surgery, South Campus, Ren Ji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 201112, China)
出处
《临床和实验医学杂志》
2018年第21期2280-2284,共5页
Journal of Clinical and Experimental Medicine
基金
上海市闵行区卫生和计划生育委员会科研课题(编号:2016MW63)
