ABSTRACT
Objective:To explore the effects of KISS1 gene transfected by lentivirus on the proliferation,invasion and migration abilities of the colorectal cancer HCT116 cells,and to clarify their mechanisms.Methods:The human colorectal cancer cells with the lowest expression level of KISS1 gene were selected.The lentiviral vectors were builted and transfected the KISS1 gene,and the cells were divided into control group (treated with PBS),empty vector group (treated with empty vector) and over-expression group(treated with KISS1 gene vector).The multiplicity of infection (MOI) of the cells was detected by fluorescence microscope.Real-time PCR and Western blotting methods were used to detect the expression levels of KISS1 mRNA and protein(metastin);CCK-8 method was used to detect the proliferation ability of the cells;Transwell chambers method was used to detect the invasion and migration abilities of the cells.Results:Among LoVo,SW620,SW480,HCT-116,and HT29 cells,the expression levels of KISS1 mRNA and protein were lowest in HCT116 cells,so they were chosen as the research carrier.After transfected with lentiviral vectors,the HCT116 cells could stably express the enhanced green fluorescent protein(EGFP) gene,and the MOI was over 80%.Compared with control group and empty vector group,the expression levels of KISS1 mRNA and protein in the cells in over-expression group were significantly increased (P0.05).Conclusion:The KISS1 gene transfected by lentivirus vector can over-express KISS1 protein and inhibit the proliferation,invasion and migration abilities of the colorectal cancer cells,and the mechanism may be related to the expression of KISS1 protein.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Kiss-1 gene suppression on the metastatic capacity of HCT116 human colorectal carcinoma cells in vitro and the involvement of nuclear factor-κB (NF-κB) signaling pathway.</p><p><b>METHODS</b>A recombinant lentiviral vector of Kiss-1 gene pGC-LV-Kiss-1-EGFP or the empty vector was transfected in HCT116 cells. Cell Counting Kit-8 (CCK8) and Transwell chamber assay were used to detect the changes in cell proliferation, invasion and migration ability after the transfection. Western blotting was used to detect the expression of I-κB, the inhibitive protein of NF-κB signal pathway, and the expression of the downstream effector MMP-9 before and after transfection.</p><p><b>RESULTS</b>In cells over-expressing Kiss-1, I-κB expression increased and MMP-9 expression decreased significantly compared to those in the blank control and vector-transfected cells (P<0.05). Kiss-1 gene over-expression resulted in significant inhibition of HCT116 cell proliferation, invasion, and migration as compared to the control cells (P<0.05).</p><p><b>CONCLUSION</b>Lentivirus-mediated Kiss-1 gene over-expression can inhibit the proliferation, invasion, and migration of HCT116 cells via the NF-B signaling pathway.</p>