| Objective We transfected a recombinant plasmid of p EGFP-N1-Kiss-1 into SW480 cells. Cells were screened with G418 to get stable expression cell lines. To investigate the relationship between Kiss-1 gene and the proliferation and migration of colon cancer cells and analyze the effect of Kiss-1 on biological chararcteristics of colon cancer, and to provide theoretic foundation for prevention and treatment of Kiss-1on colon cancer.Methods To build p EGFP-N1-Kiss-1 recombinant plasmid. The p EGFP-N1-Kiss-1 recombinant plasmid was transfected to SW480 cell by lipofectamine2000 and screened with G418. Four weeks later, cells were observed under a fluorescence microscope. The normal SW480 cells were control group, SW480 cells transfected with the p EGFP-N1 were negative control group, and SW480 cells transfected with p EGFP-N1-Kiss-1 recombinant plasmid were desired to be experimental group. Western blotting was to detect the expression of Kiss-1(Kisspeptin)in every group, CCK8 test was to test effect of Kiss-1 on proliferation of SW480 cells, migration was measured by scratch wound healing assay.Results 1. To build recombinant plasmid p EGFP-N1-Kiss-1. BLAST DNA sequencing showed that the sequence was anastomotic with Kiss-1 c DNA in Gen Bank. 2. The SW480 with stable expression of Kiss-1 was selected by G418 transfected with Kiss-1 by Lipofectamine2000. Cells were observed under a fluorescence microscope, and fluorescin was stable expression in negative control group and experimental group. The result showed that stable transfected cell lines were established. 3. Collecting protein of each group after stable transfection. The expression of Kisspeptin in experimental group was significantly higher than the control group and negative control group(P < 0.05). 4. CCK8 showed the proliferation rate of cells in the experimental group was lower than the control group and negative control group(P < 0.05). 5. Cell wound scrach assay showed: In 24 h after scracth, the healing rate of experimental group(8.1%±5.5%) was lower than that of control group(26.6%±11.4%) and negative control group(33.6%±8.7%), and the difference was significant(P < 0.05). In 48 h after scracth, the healing rate of experimental group(13.6%±5.7%) was lower than that of control group (55.5%±14.2%) and negative control group(50.5%±10.0%), and the difference was significant(P < 0.05).Conclusion Kiss-1 can inhibit the proliferation of coloretal carcinoma cells and reduce the cell migration velocity. Kiss-1 participates in control of colorectal carcinoma. |
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