Construction and inhibitory effect of MEG3 expression plasmid vector on the proliferation of human pancreatic carcinoma SW1990 cells

Abstract

Objective To construct a maternally expressed gene 3 (MEG3) expression plasmid vector, and to obtain MEG3 over-expressed human pancreatic carcinoma SW1990 cells by transfection, and to analyze 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. Key words: Pancreatic neoplasms; RNA; Gene expression regulation; Maternally expressed gene 3