摘要
目的探讨LMO4基因沉默对Snail诱导的非小细胞肺癌A549细胞上皮-间质转化(EMT)的影响。方法体外培养非小细胞肺癌A549细胞,取对数生长期细胞进行实验,根据转染慢病毒分为sh-NC组(转染sh-NC慢病毒)、sh-LMO4组(转染sh-LMO4慢病毒)、Snail组(转染Snail慢病毒)和Snail+sh-LMO4组(转染Snail慢病毒和sh-LMO4慢病毒)。采用实时定量RT-PCR检测sh-NC组和sh-LMO4组LMO4 mRNA相对表达量,采用Western Bloting法检测sh-NC组和sh-LMO4组LMO4蛋白相对表达量及4组N-cadherin、Vimentin、E-cadherin蛋白相对表达量,采用Transwell实验检测4组细胞侵袭个数,采用细胞划痕实验检测4组细胞迁移距离。结果 sh-LMO4组LMO4 mRNA相对表达量和LMO4蛋白相对表达量均低于sh-NC组(P<0.05)。Snail组侵袭细胞个数多于sh-NC组,sh-LMO4组侵袭细胞个数少于sh-NC组,Snail+sh-LMO4组侵袭细胞个数少于Snail组、多于sh-LMO4组(P<0.05)。培养24 h后Snail组细胞迁移距离长于sh-NC组,sh-LMO4组细胞迁移距离短于sh-NC组,而Snail+sh-LMO4组细胞迁移距离短于Snail组、长于sh-LMO4组(P<0.05)。Snail组N-cadherin和Vimentin蛋白相对表达量高于sh-NC组,E-cadherin蛋白相对表达量低于sh-NC组(P<0.05);sh-LMO4组N-cadherin和Vimentin蛋白相对表达量低于sh-NC组,E-cadherin蛋白相对表达量高于sh-NC组(P<0.05);Snail+sh-LMO4组N-cadherin和Vimentin蛋白相对表达量低于Snail组,E-cadherin蛋白相对表达量高于Snail组(P<0.05);Snail+sh-LMO4组N-cadherin和Vimentin蛋白相对表达量高于sh-LMO4组,E-cadherin蛋白相对表达量低于sh-LMO4组(P<0.05)。结论 LMO4基因沉默可逆转Snail诱导的非小细胞肺癌A549细胞EMT。
Objective To investigate the impact of LMO4 gene silencing on Snail- induced EMT of non- small cell lung cancer A549 cells. Methods Non- small cell lung cancer A549 cells were cultured in vitro,and logarithmic growth phase cells were extracted to carry out the experiment. According to the transfection of slow viruses,the cells were divided into A group( transfected with sh-NC slow virus),B group( transfected with sh-LMO4 slow virus),C group( transfected with Snail slow virus) and D group( transfected with Snail and sh-LMO4 slow virus). Quantitative Real- Time RT-PCR was used to detect the relative expression quantity of LMO4 mRNA of A group and B group; Western Bloting method was used to detect the relative expression quantity of LMO4 protein of A group and B group,the relative expression quantity of N-cadherin protein,Vimentin protein and E-cadherin protein of the four groups; Transwell test was used to detect the number of cell invasion of the four groups;cell scratch test was used to detect the cell migration distance of the four groups. Results The relative expression quantity of LMO4 mRNA and LMO4 protein of B group were statistically significantly lower than those of A group( P < 0. 05). The number of cell invasion of C group was statistically significantly more than that of A group,that of B group was statistically significantly less than that of A group,that of D group was statistically significantly less than that of C group,but was statistically significantly more than that of B group( P < 0. 05). After 24 hours of culture, the cell migration distance of C group was statistically significantly longer than that of A group,that of B group was statistically significantly shorter than that of A group,that of D group was statistically significantly shorter than that of C group,but was statistically significantly longer than that of B group( P< 0. 05). The relative expression quantity of N-cadherin protein and Vimentin protein of C group were statistically significantly higher than those of A group,while that
出处
《实用心脑肺血管病杂志》
2016年第5期54-58,共5页
Practical Journal of Cardiac Cerebral Pneumal and Vascular Disease
基金
国家自然科学基金(81301939)
博士后基金(2015M570697)
