Expression Of MTOR And Downstream Signal Proteins In Hela Cells Infected With Coxsackie Virus B3

Objective:To explore the protein changes of mTOR, p-mTOR, p70S6K, p-p70S6K,4EBP1, p-4EBP1of mTOR signal pathway in HeLa cells after infecting with Coxsackie virus B3.Methods:The cells were divided into two groups, which virus group was that HeLa cells were infected with CVB3, and the control group was that HeLa cells were uninfected. After the cells were cultured for3,6,9,12and24hours respectively, the Western blotting was employed to check the protein expression of mTOR, p-mTOR, p70S6K, p-p70S6K,4EBP1, p-4EBP1. Immunofluorescence was employed to check the location and expression of4EBP1.Results:1. There were significant differences in the protein expression of mTOR, p-mTOR, p70S6K, p-p70S6K,4EBP1and p-4EBP1respectively between the virus groups the control groups at different time points (all P＜0.05).2.The protein expression of mTOR and p-mTOR in virus groups was largely lower than that in control groups(P<0.05), and decreased with gradually with the prolonging of time; the protein expression of protein in control groups increased gradually with the prolongation of time.The protein expression of p70S6K and p-p70S6K in virus groups was largely lower than that in control groups(P<0.05), and decreased gradually with the prolonging of time; the protein expression of protein in control groups increased gradually with prolongation of time.The protein expression of4EBP1and p-4EBP1in virus groups was higher than that in control groups(P<0.05), and the expression of both proteins in control groups were lowest at the9h time point. The protein expression in virus groups increased gradually with the prolongation of time.There was positive correlationship between the protein expression of mTOR and the protein expression of p70S6K in virus groups, but there was no significant correlationship between these two protein expression in the control groups. And there was negative correlationship between the protein expression of mTOR with the protein expression of4EBP1in virus groups, and there was positive correlationship in the controls groups. There was positive correlationship between the protein expression of mTOR and the protein expression of p-mTOR in both groups. Also there was positive correlationship between the protein expression of p70S6K and that of p-p70S6K in virus groups, but there was no significant correlationship in the control groups. There was no correlationship between the protein expression of4EBP1and that of p-4EBP1, but there was positive correlationship in control groups.3. The expression of4EBP1at different time point could be seen mainly in cytoplasm in control groups, but the the expression of which could be seen both in cell nucleus and cytoplasm in virus groups. But the expression of4EBP1in virus groups was higher than that in control group, and expressed higher in24h time point than that in12h time point.Conclusions:1. CVB3can down-regulate the protein expression of mTOR/p70S6K signal pathway, and up-regulate the protein expression of mTOR/4EBP1signal pathway.2. CVB3can up-regulate protein expression of4EBP1as a negative regulatory growth factor.