lncRNA GHET1表达水平对人胃癌细胞MGC-803增殖迁移和侵袭的影响及其机制研究

A study on the effects of lncRNA GHET1 expression level on proliferation,migration and invasion of human gastric carcinoma cells MGC-803 and their mechanisms

目的探讨长链非编码RNA(lncRNA)胃癌高表达转录本1(GHET1)对人胃癌细胞MGC-803增殖、迁移和侵袭的影响及其机制。方法以MGC-803细胞作为实验对象,通过慢病毒转染的方法构建lncRNA GHET1过表达和lncRNA GHET1沉默的MGC-803细胞模型,分别为过表达组和沉默组,并设置其对应的阴性对照组。通过荧光显微镜观察慢病毒转染效率。通过实时荧光定量PCR检测各组lncRNA GHET1、miR-105表达水平。通过CCK-8实验检测各组细胞增殖能力。通过Transwell实验检测细胞迁移、侵袭能力。结果荧光显微镜下观察见各组细胞病毒转染率均高于90%。实时荧光定量PCR结果显示,过表达组lncRNA GHET1的相对表达量显著高于过表达对照组(P<0.05),沉默组lncRNA GHET1相对表达量显著低于沉默对照组(P<0.05),提示成功建立过表达和沉默lncRNA GHET1的MGC-803细胞模型。CCK-8实验结果显示,过表达组细胞增殖速度较过表达对照组快,而沉默组细胞增殖速度较沉默对照组慢。Transwell实验结果显示,过表达组迁移和侵袭细胞数多于过表达对照组,而沉默组迁移和侵袭细胞数少于沉默对照组,差异有统计学意义(P<0.05)。实时荧光定量PCR结果显示,过表达组miR-105表达水平显著低于过表达对照组(P<0.05),沉默组miR-105表达水平显著高于沉默对照组(P<0.05)。结论lncRNA GHET1可促进人胃癌细胞MGC-803增殖、迁移及侵袭,其机制可能与调控miR-105的表达水平有关。

Objective To investigate the effects of long non-coding ribonucleic acid(lncRNA)gastric carcinoma high expressed transcript 1(GHET1)on the proliferation,migration and invasion of human gastric carcinoma cells MGC-803 and their mechanisms.Methods MGC-803 cells were used as the experimental subjects.The MGC-803 cell models with lncRNA GHET1 overexpression and lncRNA GHET1 silencing were constructed by lentiviral transfection,which were used as the overexpression group and the silence group,respectively,and the corresponding negative control groups were set.Lentiviral transfection efficiency was observed by fluorescence microscopy.The expression levels of lncRNA GHET1 and micro ribonucleic acid-105(miR-105)in each group were detected by real-time fluorescence quantitative polymerase chain reaction(PCR).The cell proliferation ability of each group was detected by plasma cholecystokinin-8(CCK-8)assay.Cell migration and invasion abilities were detected by Transwell assay.Results Under fluorescence microscope,the viral transfection rate in the cells in each group was higher than 90%,and the results of real-time fluorescence quantitative PCR showed that the relative expression of lncRNA GHET1 in the overexpression group was significantly higher than that in the overexpression control group(P<0.05),and the relative expression of lncRNA GHET1 in the silence group was significantly lower than that in the silence control group(P<0.05),indicating that the MGC-803 cell models of the overexpressing lncRNA GHET1 and the silencing lncRNA GHET1 were successfully established.The results of CCK-8 experiment showed that the proliferation rate of the cells in the overexpression group was faster than that in the overexpression control group,while the proliferation rate of the cells in the silence group was slower than that in the silence control group.The results of Transwell experiment showed that the number of migrating and invasive cells in the overexpression group was more than that in the overexpression control group,while the number of migrating and invasive cells in the silence group was less than that in the silence control group,and the differences were statistically significant(P<0.05).The results of real-time fluorescence quantitative PCR showed that the expression level of miR-105 in the overexpression group was significantly lower than that in the overexpression control group(P<0.05),and the expression level of miR-105 in the silence group was significantly higher than that in the silence control group(P<0.05).Conclusion lncRNA GHET1 can promote the proliferation,migration and invasion of human gastric carcinoma cells MGC-803,and their mechanisms may be related to the regulation of the expression level of miR-105.