体外沉默Rce1对舌鳞状细胞癌细胞侵袭和迁移能力的影响
Effects of silencing Rce1 in vitro on the invasion and migration of tongue carcinoma
作者:孙君君,陶匀雅,周元,何宗轩,盛善桂,王奇民,童磊,赵开,王少如,陈正岗
Author:Junjun Sun,Yunya Tao,Yuan Zhou,Zongxuan He,Shangui Sheng,Qimin Wang,Lei Tong,Kai Zhao,Shaoru Wang,Zhenggang Chen
收稿日期:2018-06-11 年卷(期)页码:2019,37(2):143-143-148
期刊名称:华西口腔医学杂志
Journal Name:West China Journal of Stomatology
关键字:Rce1,鳞状细胞癌,侵袭,迁移,
Key words:Rce1,squamous cell carcinoma,invasion,migration,
基金项目:国家自然科学基金(81372908)
中文摘要
目的 通过RNA干扰技术沉默Rce1基因,探讨Rce1对舌癌细胞侵袭和迁移能力的影响。方法 体外培养舌鳞状细胞癌Cal-27和SCC-4细胞,设计合成Rce1基因的小干扰RNA(siRNA),采用脂质体载体瞬时转染沉默Rce1基因。根据转染的siRNA不同,将实验组分为Rce1-siRNA-1组、Rce1-siRNA-2组、Rce1-siRNA-3组,脂质体载体分别转染相对应序列的siRNA。脂质体转染siCON作为阴性对照组,不转染siRNA作为空白对照组。采用实时定量聚合酶链反应检测各组Rce1、RhoA以及K-Ras基因的表达,蛋白质印迹法(Western blot)检测Rce1、RhoA、K-Ras、金属基质蛋白酶(MMP)-2及MMP-9的表达,采用体外侵袭实验检测Cal-27和SCC-4的侵袭能力,细胞划痕实验检测细胞迁移能力。结果 实时定量聚合酶链反应及Western blot检测结果显示,与阴性对照组和空白对照组相比,实验组的Rce1基因及蛋白表达均下调(PP>0.05),MMP-2、MMP-9表达下降(PPP结论 体外沉默Rce1可以有效下调其在舌癌细胞Cal-27和SCC-4中的表达水平,降低迁移和侵袭能力。
英文摘要
ObjectiveThis study aimed to explore the influence of Rce1 on invasion and migration of tongue squamous cell carcinoma cells by silencing the Rce1 gene with RNA interference.MethodsThe tongue squamous cell carcinoma Cal-27 and SCC-4 cells were culturedin vitro. The small interfering RNA (siRNA) of the Rce1 gene was designed, and the Rcel gene expression was silenced vialiposome transfection. According to the siRNA transfected by liposome, the experimental group was divided into three groups, namely, Rce1-siRNA-1, Rce1-siRNA-2, and Rce1-siRNA-3 groups. Negative control group was transfected by siCON, and the blank control group was untransfected by siRNA. The Rce1, RhoA, and K-Ras gene expression levels in each group were analyzed by real-time quantitative polymerase chain reaction. The Rce1, RhoA, K-Ras, MMP-2, and MMP-9 protein expression levels were analyzed by Western blot. The invasiveness of tongue cancer cell Cal-27 and SCC-4 were determined by Transwell invasion assay, and cell migration assay was performed by cell scratch assay.ResultsReal-time quantitative polymerase chain reaction and Western blot results showed that compared with the negative and blank control groups, the Rce1 gene and protein expression levels in three experimental groups decreased (PP>0.05). Meanwhile, the MMP-2 and MMP-9 expression levels decreased (PPPConclusionSilencing Rce1in vitrocan effectively downregulate its expression in tongue squamous cell carcinoma cells Cal-27 and SCC-4 and reduce the migration and invasion abilities of these cells.
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