The Experimental Research On Effect Of AiKang-Fang On Human Lung Adenocarcinoma A549 Cell、Mouse Lewis Cell PI3K/AKT Singnal Pathway

Objective Group preliminary experiments confirmed Ai KangFang serum on A549 cell and Lewis cell growth inhibition and apoptosis effect obviously,based on our experiments following the previous study at best 20% medicated serum on the cell culture,to observe Ai Kang-Fang medicated serum role in human lung adenocarcinoma A549 cells,mice Lewis phosphatidyl inositol 3 kinase(PI3K),protein kinase-B(Akt)and phosphorylated protein kinase-B(p-Akt)protein expression and the influence of the mRNA level of PI3 K and AKT,and then discusses whether the party through PI3K/Akt signaling pathways play a role of apoptosis.Methods The experimental method for the preparation of serum containing drugs was prepared in the previous stage,80 SD rats group: normal saline control group(NS group),Ai Kang-Fang low,medium and high dose group,cyclophosphamide(CTX)therapy group,back Ai Kang-Fang is low,medium and high doses respectively combined cyclophosphamide chemotherapy group(low dose + CTX,dose + CTX,high dose + CTX),10 in each group,the preparation of medicated serum.The cell culture was grouped according to the serum containing the drug: the blank control group(only medium),control group and experimental group and control group for NS group,experimental group for the low dose group,middle dose group,high dose group,CTX group,low dose + CTX group,middle dose + CTX group,high dose +CTX group.Ai Kang-Fang different doses,cyclophosphamide and combined cyclophosphamide drug-containing serum respectively different doses on cultivating A549 and Lewis cells,using Immunocytochemistry and Westorn Blot methods to test PI3 K,Akt,and p-Akt protein expression and RT-qPCR method to detect PI3 K,and Akt mRNA level.Results1.Effect of Ai Kang-Fang on human lung adenocarcinoma A549 cells PI3 K and AKT expression.Immunocytochemistry method and the Westorn Blot combined detection of PI3 K,and AKT protein expression levels: each time the experimental cell PI3 K,and AKT protein expression were lower than the blank control group,NS group(P<0.05),the experimental group protein expression of successive substantially reduced(P<0.05),significantly less than24 hours of protein expression 48 hours,the results of Westorn Blot test for p-AKT 24 showed that,compared with the blank control group and the NS group,the high dose,CTX group and low,middle and high combined CTX group protein were significantly lower than that in the control group(P<0.05);Rt-qPCR detected the levels of PI3 K and Akt mRNA: the mRNA level of the experimental group at each time period was low and the blank control group;The results showed that the overall trend was significantly lower when the results were compared with 24 hours,and the comparison between the experimental groups was generally decreased.2.The influence of Ai Kang-Fang on the expression of PI3 K and AKT in mouse Lewis cells.Immunocytochemistry method and the Westorn Blot combined detection of PI3 K,and AKT protein expression levels: each time the experimental cell PI3 K,and AKT protein expression were lower than the blank control group,NS group(P<0.05),the experimental group protein expression of successive substantially reduced(P< 0.05),significantly less than24 hours of protein expression 48 hours,and then,the results of Westorn Blot test for p-AKT24 hours showed that there was a down-regulation trend of low,medium,high and different doses of Ai Kang-Fang(P<0.05).The low protein expression was most obvious in the medium dose +CTX group;Rt-qPCR detected the levels of PI3 K and Akt mRNA: the mRNA level of the experimental group at each time period was low and the blank control group;Theresults of 48 hours were less than those of 24,and the comparison between the experimental groups was generally decreased.Conclusions1.The synergistic cyclophosphamide of Ai Kang-Fang and Ai Kang-Fang can reduce the expression of PI3 K,AKT and p-Akt protein in the human lung adenocarcinoma A549 cells and mouse Lewis cells,and the effect of the effect is certain time and dose dependence.2.The synergistic cyclophosphamide between Ai Kang-Fang and Ai Kang-Fang decreased the level of PI3 K and AKT mRNA in the human lung adenocarcinoma A549 cells and mouse Lewis cells.3.Ai Kang-Fang may be possible to promote cell apoptosis by inhibiting the PI3K/AKT signal transduction pathway,which may be one of the ways to fight non-small cell lung cancer.