Effect Of Decorin On Molecular Biology Behavior Of Human Gastric Cancer Cells

Objective: To construct PEGFP-N1-DCN eukaryotic expression vector and transfect into human gastric cancer cell lines SGC-7901 to observe its effect on cell proliferation, cell cycle, cell apoptosis, migration and invasion.Methods: (1) Total mRNA was extrcted from humam placenta tissue and the DCN cDNA was obtained by RT-PCR and cloned into pEGFP-N1 vetor, and then the pEGFP-N1-DCN vector was constructed, identified and transfected into the Gastric cancer cells SGC-7901. Its expression was detected by by fluorescent microscopy, RT-PCR and Western blot analysis. (2) SGC-7901 transfected by pEGFP-N1-DCN by liposome, the expression of DCN, EGFR and TGF-β1 were detected by Wester blot and RT-PCR. (3) Proliferation of cells detected by MTT; cell cycle and apoptosis detected by flow cytometry. (4) Effective of cell migration and invasion dectected by transwell tests.Results: (1) Enzyme digestion analysis and sequencing showed that the target genes coloned by RT-PCR was found to be 1080bp. (2) Green fluorescence was emitted from transfected cells under fluorescent microscope. RT-PCR and Western blot demonstrated that the recombinant vector pEGFP-N1-DCN was expressed in SGC-7901 cells, and the relatived molecular mass of the GFP-DCN fusion protein was about 67×10~3, while the relatived molecular mass of the GFP protein expressed by pBud-GFP was about 27×10~3. After SGC-7901 transfected by pEGFP-N1-DCN, the expression of EGFR and TGF-β1 were decreased. (3) MTT demonstrated that cell proliferation was inhibited; flow cytometric analysis found that the cells arrested in G0~G1 phase and the rate of apoptosis was increased. (4)The migration and invasion ability of cells were evidently decreased.Conclusion: After pEGFP-N1-DCN transfected into SGC-7901 cells, proliferation was significantly inhibited, cell cycle arrested in G0~ G1 phase, the rate of apoptosis was increased and the ability of migration and invasion were decreased. It can be see that DCN play a role of anti-tumor by inhibiting tumor cell proliferation, migration and invasion and promoting apoptosis.