Construction and inhibitory effect of MEG3 expression plasmid vector on the proliferation of human pancreatic carcinoma SW1990 cells
10.3760/cma.j.issn.1673-422X.2015.07.003
- VernacularTitle:人 MEG3基因质粒载体的构建及其对人胰腺癌SW1990细胞增殖的影响
- Author:
Ruidong CHEN
;
Wen TANG
;
Duanmin HU
- Publication Type:Journal Article
- Keywords:
Pancreatic neoplasms;
RNA;
Gene expression regulation;
Maternally expressed gene 3
- From:
Journal of International Oncology
2015;(7):488-491
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a maternally expressed gene 3(MEG3)expression plasmid vec-tor,and to obtain MEG3 over-expressed human pancreatic carcinoma SW1990 cells by transfection,and to ana-lyze the effect of MEG3 overexpression on the proliferation of human pancreatic carcinoma SW1990 cells. Methods A complete gene sequence based on the sequence of MEG3 in the GenBank was designed and inser-ted into the eukaryotic expression vector pcDNA3. 0 to construct recombinant plasmid pcDNA3. 0-MEG3. It was identified by sequencing and transfected into human pancreatic carcinoma SW1990 cells. The expression of MEG3 in SW1990 cells was confirmed by RT-PCR. The effect of MEG3 on proliferation was evaluated by MTT assay. In this study,the SW1990 cells transfected by plasmid pcDNA3. 0 were named negative control group, and the usual SW1990 cells were named blank control group. Results A MEG3 expression plasmid vector-pcDNA3. 0-MEG3 was constructed successfully. And pcDNA3. 0-MEG3 vector was transfected into SW1990 cells successfully. The expression of MEG3 at mRNA in MEG3-SW1990 cells increased significantly,about 895 times(F = 73. 592,P ﹤ 0. 01). The results of MTT assay indicated that over-expressed MEG3 could obviously inhibit SW1990 cells proliferation in vitro. After SW1990 cells transfected with pcDNA3. 0-MEG3 for 72 hours, the absorbance value was 0. 81 ± 0. 06,with a statistically significance(F = 33. 489,P ﹤ 0. 01)compared with negative control group(1. 17 ± 0. 07)and blank control group(1. 08 ± 0. 03). Conclusion A MEG3 expre-ssion plasmid vector-pcDNA3. 0-MEG3 is constructed successfully. It is confirmed that MEG3 and its product have obvious inhibitory effects for the proliferation of human pancreatic carcinoma SW1990 cells.
