Regulation Effects Of CVB3 On Hela Cell MTOR/70S6K Signal Pathway Expression

Objectives:This study employed CVB3 to infect "no serum starvation" and "serum starvation" Hela cell to observe cytopathinc effection and detect the mRNA content of coxsackie virus, mammalian target of rapamycin, ribosomal protein S6 kinase at different time. To explore the effect of cell cycle to mTOR signal pathway and detect the expression of mTOR, p70S6K mRNA on Hela cell.Methods:The study included virus group and control group. Virus group employed CVB3 to infect "no serum starvation" and "serum starvation" Hela cell at 3 h,6 h,9 h,12 h,24 h. Control group was cultured by 2% fetal bovine serum. Cell morphology change was observed by phasecontrast microscope and the expression of mTOR, p70S6K mRNA was detectd with RT-PCR and Real-Time PCR on Hela cell at different time. SPSS 18.0 software was used for the statistical analysis. P<0.05 was considered to be statistically significant.Results:1. Observed cell morphology change that cell was cultured with two different methods:The virus group has cytopathinc at the 6th h on"no serum starvation" and "serum starvation" Hela cell; and the longer time effected, the more cytopathinc effection.2. The expression of CVB3 mRNA on Hela by RT-PCR and Real-Time PCR:The results of RT-PCR and Real-Time PCR all suggested that the virus group on "no serum starvation" and "serum starvation" Hela cell of the expression of CVB3 mRNA were increased in time dependence. The "no serum starvation" Hela cell compared with "serum starvation" Hela cell had no statistically significant (P>0.05).3. The expression of mTOR, p70S6K mRNA on Hela cell by RT-PCR and Real-Time PCR:The virus group on "no serum starvation" Hela cell of the expression of mTOR, p70S6K mRNA were noticeable down-regulation at the 12th h, which compared with the control group had statistically significant (P<0.05). However, the virus group on "serum starvation" Hela cell of the expression of mTOR, p70S6K mRNA were all lower than that of the control group (all P<0.05); The virus group in two different cultured Hela cell had no expression of mTOR and p70S6K mRNA at the 24th h by RT-PCR, but extremely expression of mTOR and p70S6K mRNA was observed by Real-Time PCR.4. "No serum starvation" Hela cell compared with "serum starvation" Hela cell in the expression of mTOR, p70S6K mRNA:The results of RT-PCR and Real-Time PCR all suggested that the "serum starvation" virus group and the control group on Hela cell of the expression of mTOR mRNA were all higher than that of "no serum starvation" virus group (all P<0.05); However, the "serum starvation" virus group and the control group on Hela cell of the expression of p70S6K mRNA were all higher than "no serum starvation" virus group, but they had no statistically significant (all P<0.05)Conclusions:1. The "no serum starvation" and the "no serum starvation" virus group on Hela cell of the expression of mTOR, p70S6K mRNA were all lower than that of the control group, which corresponded that CVB3 maybe down-regulate the expression of mTOR, p70S6K mRNA of Hela cell.2. The expression of CVB3 mRNA was up-regulated when CVB3 infected Hela cell at the 12th and the 24th h, but the expression of mTOR, p70S6K mRNA were down-regulated. CVB3 mRNA had high expression when CVB3 infected Hela cell at the 24th h, however, had no or extremely the expression of mTOR, p70S6K mRNA, suggested that mTOR signal pathway might be take part in the CVB3-induced apoptosis on Hela cell.3. The "no serum starvation" virus group on Hela cell of the expression of mTOR, p70S6K mRNA which compared with the control group had statistically significant only at the 12th and the 24th h, however, The "serum starvation" virus group had statistically significant at all different time. And the "serum starvation" virus group and the control group on Hela cell of the expression of mTOR mRNA were all higher than "no serum starvation" virus group, which suggested that the different cell cycle had taken effect on mTOR signal pathway, and "serum starvation" maybe reduce the experiment influence because of cell cycle and better than "no serum starvation".