Overexpression Of Kiss-1 Reduces Colorectal Cancer Cell Proliferation, Migrate, Invasion And Promote Cell Apoptosis By Blocking PI3K/AKT/NF-?B-/MMP-9 Signal Pathway

[Object]1.To further evaluate the function of metastasis-suppressor Kiss-1, which regulates cell growth, migrate and invasion in colorectal cancer cells.2.To clarify Kiss-1 control cell mobility, chemotaxis and invasion of colorectal cancer by reducing MMP-9 via blocking PI3K/AKT/NF-kB signal pathway.[Method]1.HCT116 cells were infected with recombinant lentivirus, and expression of Kiss-1 in infected cells was verified by quantitative reverse transcription PCR(qRT-PCR) and immunoblotting. Cell proliferation was analyzed by Cell Counting Kit-8(CCK-8); Flow cytometry was used to determine cell apoptosis and cells was marked by Annexin V-APC/7-AAD. Cell migrate and invasion were studied by Transwell assays(with and without Matrigel). Expression level of MMP-9 in HCT116 cells used immunoblotting.2.Protein expression of molecules in PI3K/AKT/NF-kB/MMP-9 pathway were determined in mock, negative control and high-expressed Kiss-1 group cells by immunoblotting.3.Activation of PI3 K pathway in high-expression of Kiss-1 group cells used 740Y-P(PI3K activator) and expression of PI3 K, AKT, pAKT, NF-kB-P65 and MMP-9 were detected by immunoblotting. Simultaneously, we also detected cell proliferation, cell apoptosis, cell migrate and invasion.4.Activation of AKT pathway in high-expression of Kiss-1 group cells used PDGF(AKT activator) and expression of PI3 K, AKT, pAKT, NF-k B-P65 and MMP9 were detected by immunoblotting. Simultaneously, we also detected cell proliferation, cell apoptosis, cell migrate and invasion.[Results]1.We obtained stable cells with high or low expressed Kiss-1 by lentivirus system in HCT116 cells. Overexpression of Kiss-1 inhibited significantly cell proliferation in HCT116 cells(P<0.01). However, Silencing Kiss-1 in HCT116 cells could not affect cell proliferation. In addition, high-expressed Kiss-1 decreased significantly expression of MMP-9 in HCT116 cells, which compare to control group. But low-expressed Kiss-1 didn't increased expression of MMP-9. Exogenous expression of Kiss-1 promoted cell apoptosis(p<0.01) in HCT116 cells, and knock-out Kiss-1 in HCT116 cells could not improve anti-apoptotic capacity.2.Exogenous expression of Kiss-1 inhibited expression of PI3 K, pAKT, NF-kB-P65 and MMP-9, and expression of AKT maintain unchanged in HCT116 cells.3.After 740Y-P(PI3K activator) treated in high-expressed Kiss-1 HCT116 cells, expression of PI3 K, pAKT, NF-kB-P65 and MMP-9 were increased and expression of AKT maintain unchanged, cell proliferation(p<0.01), anti-apoptotic(p<0.01), cell migrate and invasion(p<0.001) were promoted, which compared to high-expressed Kiss-1 HCT116 cells.4.After PDGF(AKT activator) treated in high-expressed Kiss-1 HCT116 cells, expression of pAKT, NF-kB-P65 and MMP-9 were increased, expression of PI3 K and AKT maintain unchanged, cell proliferation(p<0.01), anti-apoptotic(p<0.01), cell migrate and invasion(p<0.001) were promoted, which compared to high expressed Kiss-1 HCT116 cells.[Conclusion]1.Overexpression of Kiss-1 reduces cell proliferation, and promotes cell apoptosis in colorectal cancer HCT116 cells.2.In colorectal cancer HCT116 cells, overexpression of Kiss-1 suppresses colorectal cancer cell metastasis and invasion by reducing expression of MMP-9 via blocking PI3K/AKT/NF-?B signal pathway.