| Objective: To investigate the antitumor activity of interleukin-18 gene lentiviral expression vector on lung adenocarcinoma cell line A549 in vitro in order to provide a new way for the prevention and treatment of lung cancer. Methods: The transfection of human interleukin-18 gene lentiviral expression vector of human lung adenocarcinoma A549 cells identified as IL-18 intervention group(group A),only transfected with lentiviral expression vector without containing human interleukin-18 gene in human lung adenocarcinoma A549 cells identified as lentiviralempty vector group(group B),without any intervention of human lung adenocarcinoma A549 cells identified as blank control group(group C).The expressing of IL-18 of 3 groups for 24 h and 96 h were tested by western bloting.The growth curve of 3 groups from 24 h to 96 h were described by MTT assay.The apoptosis rate and cell cycle of 3 groups for 24 h was analyzed by flow cytometry.The expressing of the Th1 cell subsets cytokine interferon-γand Th2 cell subsets cytokine interleukin-4 of 3 groups for 24 h and 96 h were tested by ELISA. Resμlts: The resμlts of Western Blotting showed that the expression of IL-18 in group A were significantly increased than group B and group C after 96 h cμlture(P<0.05),but no statistical significance after 24h(P>0.05).According to the resμlt of MTT, group A cell proliferation ability were significantly higher than group B and group C.According to the resμlt of MTT growth curve,group A growth at the same time reduction,compared to group B and group C,however group B and group C were basically the same.After 96 h of cμlture, the apoptosis rate of group A were significantly higher than that of group B and group C(P<0.05),however the differences of the apoptosis rate of group B and group C showed no significance(P>0.05).group A of cells in G0 phase,G1 phase were significantly increased than group B and group C(P <0.05),and group A of cells in S,M phase were significantly reduced than group B and group C(P <0.05),with no significant difference in group B and group C(P>0.05).ELISA resμlts showed that the expression of IFN- γ of group A were higher than group B and group C(all P<0.05),the expression level of IL-4 in group A were significantly less than that group B and group C(all P <0.05),but there were no statistical significance among 3 groups about IFN- γ /IL-4 ratio(P >0.05),and the expression of IFN- γ,IL-4,IFN- γ /IL-4 in group B and group C were no statistical significance(P >0.05),after 24 h of cμlture.And after 96 h of cμlture,the expression of IFN- γ in group A were significantly higher than group B and group C(P<0.05),the expression of IL-4 in group A were significantly reduced than group B and group C(P<0.05),the IFN- γ /IL-4 ratio in group A were significantly higher than group B and group C(P<0.05),the expression of IFN-γ,IL-4,IFN- γ /IL-4 in group B and group C were no statistical significance(P >0.05). Conclusions: The interleukin-18 gene lentiviral expression vector can stable and effective express IL-18,can diminished lung adenocarcinoma A549 cell proliferation ability,inhibit growth of tumor cell, enhanced tumor apoptosis,promoted the secretion of Th1 cytokines IFN- γ,decreased the producing of Th2 cytokines IL-4 correspondly, reverse the imbalance of Th1/Th2 cell subsets,and maintain the balance of Th1/Th2 cell subsets,thus play an active anticancer effect,expected to open up a new way of prevention and control of lung cancer. |
