摘要
目的探讨siRNA干扰ALDH1A1基因表达对鼻咽癌5-8F和CNE2细胞迁移侵袭能力的影响。方法将鼻咽癌5-8F和CNE2细胞分为5-8F干扰组、5-8F空载组、CNE2干扰组、CNE2空载组,利用已构建的慢病毒pMagic 4.1-ALDH1A1siRNA载体和pMagic 4.1-shRNA空载体分别感染5-8F和CNE2细胞,建立稳定干扰ALDH1A1表达的鼻咽癌5-8F、CNE2细胞株和空载的5-8F和CNE2细胞株。4组采用Western blot法检测细胞ALDH1A1、血管内皮生长因子(vascular endothelial growth factor,VEGF)、基质金属蛋白酶(matrix metalloproteinase,MMP)-2、MMP-9蛋白表达,采用Transwell小室实验检测各组迁移细胞数,采用Boyden小室实验检测各组侵袭细胞数。结果 5-8F干扰组ALDH1A1蛋白表达(0.34±0.01)低于5-8F空载组(0.75±0.03),CNE2干扰组ALDH1A1蛋白表达(0.24±0.03)低于CNE2空载组(0.71±0.03);5-8F干扰组鼻咽癌细胞迁移细胞数(35.0±2.6)、侵袭细胞数(32.7±2.1)均低于5-8F空载组(117.6±3.2,105.6±4.0),CNE2干扰组鼻咽癌细胞迁移细胞数(34.0±3.0)、侵袭细胞数(32.3±1.5)均低于CNE2空载组(121.0±4.0,108.0±1.7)(P<0.01);5-8F干扰组VEGF(0.49±0.03)、MMP-2(0.36±0.04)、MMP-9(0.31±0.04)蛋白表达低于5-8F空载组(0.73±0.02、0.50±0.02、0.89±0.01)(P<0.01),CNE2干扰组VEGF(0.48±0.02)、MMP-2(0.37±0.02)、MMP-9(0.41±0.02)蛋白表达低于CNE2空载组(0.78±0.05、0.67±0.05、0.76±0.04)(P<0.01)。结论下调ALDH1A1基因表达可抑制鼻咽癌细胞迁移侵袭。
Objective To investigate the effect of ALDH1A1 silence on the migration and invasion of nasopharyngeal carcinoma cell lines 5-8F and CNE2. Methods The nasopharyngeal carcinoma 5-8F and CNE2 cell lines were divided into 5-8F shRNA-ALDH1A1 group, 5-8F control group, CNE2 shRNA-ALDHIA1 group, and CNE2 control group. The lentiviral pMagic 4. 1-ALDHIA1 siRNA and pMagic 4. 1-shRNA were used to transfect 5-8F and CNE2 cell lines respectively to construct 5-8F, CNE2, 5-8F vector and CNE2 vector cell lines stably interfering the expression of ALDH1A1. The protein expression of ALDH1A1, vascular endothelial growth factor （VEGF）, matrix metalloproteinase （MMP）-2 and MMP-9 were evaluated with Western blot in four groups. The counts of migration and invasion cells were detected by Transwell chamber and Boyden chamber assay. Results The expression of ALDH1A1 was significantly lower in 5-8F shRNA-ALDH1A1 group （0. 34±0. 01） than that in 5-8F vector group （0. 75 ± 0. 03）, and in CNE2 shRNA-ALDH1A1 group （0.24±0.03） than that in CNE2 vector group （0.71±0.03） （P〈0.05）. The migration and invasion cell counts were significantly lower in 5-8F shRNA-ALDHIA1 group （5.0 ± 2.6, 32.7 ± 2.1） than those in 5-8F vector group （117.6±3.2, 105.6d=4.0）, and in CNE2 shRNA-ALDH1A1 group （34.0±3.0, 32.3±1.5） than those in CNE2 vector group （121. 0±4. 0, 108. 0±1. 7） （P〈0.01）. The expressions of VEGF, MMP-2 and MMP-9 were significantly lower in 5-8F shRNA-ALDH1A1 group （0.49±0.03, 0. 36±0. 04, 0. 31±0. 04） than those in 5-8F vector group （0.73±0.02, 0.50±0.02, 0.89±0.01） （P〈0.01）, in CNE2 shRNA-ALDH1A1 group （0.49±0.03, 0.36±0.04, 0. 31±0. 04） than those in CNE2 vector group （0.78±0.05, 0. 67±0. 05, 0. 76±0. 04） （P〈0.01）. Conclusion The down-regulation of ALDH1A1 could inhibit the migration and invasion of nasopharyngeal carcinoma cells.
作者
吴爱兵
李金媚
罗怡平
麦宗炯
杨志雄
WU Ai-bing;LI Jin-mei;LUO Yi-ping;MAI Zong-jiong;YANG Zhi-xiong(Department of Oncology , the Affiliated Hospital of Guangdong Medical University, Zhanjiang 524000, Chin)
出处
《中华实用诊断与治疗杂志》
2018年第4期327-330,共4页
Journal of Chinese Practical Diagnosis and Therapy
基金
国家自然科学基金青年基金(81201672)
广东省科技计划项目(2016A020215228)
广东医学院博士启动基金(B2012032)
关键词
鼻咽癌
乙醛脱氢酶A1
侵袭迁移
Nasopharyngeal carcinoma, ALDHIAI
migration and invasion
