过表达LncRNA-MEG3对肠癌细胞Lovo增殖活性的影响  

Effect of LncRNA-MEG3 over-expression on the proliferation activity of colorectal cancer Lovo cells

作　　者：章杰兵[1] 徐燕茹[2] 邱彦[3] 霍中华[3] 

机构地区：[1]南京军区联勤部药品仪器检验所,南京210002 [2]南京军区卫生部,南京210002 [3]解放军第四五四医院

出　　处：《海军医学杂志》2014年第5期349-351,367,共4页Journal of Navy Medicine

摘　　要：目的用PCR扩增长链非编码RNA-MEG3,构建MEG3真核表达载体,将重组质粒转染人肠癌细胞Lovo,观察MEG3含量改变对Lovo细胞增殖活性的影响。方法用293细胞提取人总RNA,反转录制备cDNA,PCR扩增MEG3基因并构建克隆,阳离子脂质体法转染重组质粒至Lovo细胞,转染后48 h通过荧光标记物GFP观察细胞转染效率。RealTime-PCR检测转染后细胞内MEG3含量变化。CCK-8检测肿瘤细胞对数生长期基因干预对于增殖活性的影响。结果成功获取了MEG3基因并构建了重组表达载体;成功转染Lovo细胞,转染后48 h,细胞转染效率约为60%;转染后细胞内MEG3含量明显升高,与空质粒转染组比较,相对含量上升约6.8倍,差异有统计学意义(P<0.01);外源MEG3的高表达可抑制Lovo细胞增殖活性,基因干预后48、72 h,与未转染组及转染对照组细胞比较,差异有统计学意义(P<0.01)。结论 LncRNA-MEG3可显著抑制Lovo细胞增殖活性。Objective To investigate the effect of the changes in MEG3 level on proliferation activity in Lovo cells,through the amplification of the non-coding RNA-MEG3 by PCR and the construction of a eukaryotic expression vector for MEG3,which was transfected into a human colon cancer cell line,Lovo. Methods cDNA was prepared from the total RNA extracted from 293 cells by reverse transcription and MEG3 gene was amplified by PCR and was used to construct a recombinant plasmid,which was transfected into Lovo cells by using cationic liposome. Transfection efficiency was evaluated by observation on the expression of GFP marker 48 hours after transfection,and changes in MEG3 content were detected by RT-PCR（ Real-time-PCR）. CCK-8 was used to measure the effect of genetic intervention on proliferative activity in tumor cells in the logarithmic growth phase. Results MEG3 gene was successfully obtained and the recombinant expression vector was constructed. Lovo cells were successfully transfected,and 48 hours after transfection,the transfection efficiency reached as high as 60%. MEG3 level in transfected cells was significantly increased for approximately 6. 8 folds,as compared with that of the transfected control group（ P〈0. 01）. High expression of exogenous MEG3 in Lovo cells could inhibit cell proliferation,and significant differences could be noted at hours 48 and 72 after the genetic intervention,as compared with those of the untransfected group and the transfected control group（ P〈0. 01）. Conclusion LncRNA-MEG3 could obviously inhibit the proliferation of Lovo cells.

关 键 词：LncRNA LOVO MEG3 细胞活性 

分 类 号：R735.3[医药卫生—肿瘤]