Effect of LncRNA-MEG3 over-expression on the proliferation activity of colorectal cancer Lovo cells
10.3969/j.issn.1009-0754.2014.05.005
- VernacularTitle:过表达 LncRNA-MEG3对肠癌细胞 Lovo增殖活性的影响
- Author:
Jie-Bing ZHANG
1
;
Yan-Ru XU
;
Yan QIU
;
Zhong-Hua HUO
Author Information
1. 南京军区联勤部药品仪器检验所
- Keywords:
LncRNA;
Lovo;
MEG3;
Proliferation activity
- From:
Journal of Navy Medicine
2014;(5):349-351,367
- CountryChina
- Language:Chinese
-
Abstract:
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 transfect-ed into a human colon cancer cell line, Lovo.Methods cDNA was prepared from the total RNA extracted from 293 cells by reverse tran-scription 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.
