MEG3对鼻咽癌细胞转移能力的影响及相关机制的初步探究

Effect of MEG3 on metastasis ability of nasopharyngeal carcinoma cells and preliminary study of related mechanisms

目的探究长链非编码RNA母系表达基因3(maternally expressed gene3,MEG3)对鼻咽癌细胞侵袭迁移能力的影响及相关机制。方法慢病毒转染法在人鼻咽癌细胞CNE1中构建过表达MEG3的实验组细胞及转染空白载体的对照组细胞,荧光定量PCR(quantitative real-time PCR,RT-q PCR)技术检测两组细胞MEG3表达水平,Transwell实验检测两组细胞侵袭迁移能力变化,免疫印迹技术(Western Blot)检测两组细胞上皮间充质转化(epithelial-mesenchymal transition,EMT)相关分子E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)及β-链蛋白(β-catenin)表达水平。结果实验组细胞MEG3表达显著高于对照组细胞,差异具有统计学意义(P<0.01)。Transwell侵袭实验结果示实验组细胞穿透基底膜的细胞数显著低于对照组细胞,Transwell迁移实验结果示实验组细胞迁出小孔细胞数显著低于对照组细胞,差异具有统计学意义(P<0.05);Western Blot实验结果示实验组相比对照组细胞E-cadherin表达水平显著上调,而Vimentin及β-catenin表达水平显著下调,差异具有统计学意义(P<0.05)。结论 MEG3可下调人鼻咽癌细胞CNE1的侵袭迁移能力,其机制与抑制细胞EMT进程有关。

Objective To investigate the effect of long non-coding RNA maternal expressed gene 3（ MEG3）on invasion and migration ability of nasopharyngeal carcinoma cells and the related mechanisms. Methods Lentiviral transfection was used to construct overexpressing MEG3 cells as experimental group,and transfected with empty vector as control group in human nasopharyngeal carcinoma CNE1 cells. Quantitative Real-time PCR（ RT-q PCR） technology was used to detect the expression level of MEG3 in two groups of cells. Transwell assay was used to detect the ability of migration and invasion in two groups of cells. Western blot technique was used to detect the expression level of epithelial mesenchymal transition（ EMT） related molecules including E-cadherin,Vimentin and β-catenin. Results The expression of MEG3 in the experimental group was significantly higher than that in the control group,and the difference was statistically significant（ P〈0. 01）. The results of Transwell invasion assay showed that the number of cell through the basement membrane in the experimental group cells was significantly lower than that in the control group,and the results of Transwell migration assay showed that the number of cells through the holes in the experimental group cells was also significantly lower than that in the control group,and the difference was statistically significant（ P〈0. 05）. The results of Western Blot showed the expression of E-cadherin in the experimental group was increased,and the expression of Vimentin and β-catenin in the experimental group was significantly reduced compared to the control group,and the difference was statistically significant（ P〈0. 05）. Conclusion MEG3 can decrease the invasion and migration ability of human nasopharyngeal carcinoma cell CNE1,and its mechanism is related to the inhibition of cell EMT progression.