Influence of transfection of CD40L gene on cell proliferation and invasion of colon cancer cells

Objective:CD40/CD40 ligand(CD40L) interaction plays an essential role in cell-mediated anti-tumor immune response.However,the role of CD40L in mouse colon cancer gene therapy is not clear.We aimed to investigate the impacts of CD40L gene on the biological features such as proliferation and invasion capabilities of colon26 cells and discuss the feasibility of using CD40L as a molecular target for colon cancer gene therapy.Methods:The pMKITneo-CD40L plasmids were transfected into mouse colon26 cells mediated by LipofactamineTM2000.The mRNA transcription of CD40L was analyzed by RT-PCR.Western blot and laser scanning confocal microscopy were used to detect the CD40L protein expression.MTS method was used to determine the effect of CD40L gene transfection on cell growth;the phenotype of CD40L cells was assessed using flow cytometry;the expression of matrix metalloproteinase 2(MMP-2) mRNA was analyzed by RT-PCR;the invasion of cells was detected with Transwell chamber invasive assay.Results:RT-PCR indicated that the level of CD40L mRNA expression was(1.09±0.12) after transfection with CD40L gene,which was increased by 59.21% and 58.33% compared with the cells transfected with empty vector(0.69±0.08) and the non-transfection group(0.69±0.11,P0.05).The level of MMP-2 mRNA was significantly decreased in colon26/CD40L cells.MTS showed that there was no significant difference in the growth speed between colon26/CD40L cells and parent colon26 cells.The expression levels of CD80 and CD86 were higher in colon26/CD40L cells than those on parent cells.The result of Transwell assays suggested that the invasion ability of colon26/CD40L cells decreased significantly(P0.05).Conclusion:The immunogenicity of colon26/CD40L cells are enhanced and the invasion capability is decreased.The malignant phenotype of colon26 cells is inhibited after CD40L gene transfection.