The Effect Of PTH On Cbfα1 Expression In MC3T3-E1 Cell Differentiation Process

Objective:To study the Cbfα1 expression when stimulating MC3T3-E1 cells with hPTH1-34 Intermittencely and continually.To discuss the molecular mechanisms of hPTH1-34 on bone formation;Finally,to provide the experimental basis for the hPTH1-34 treatment of osteoporosis.Method:(1)Cell's transfer of generation raise(2)Use the classical method to induce the MC3T3-E1 cell,in the 1,3,6,9,12,15 days MC3T3-E1 cell,carries on semi-quantitative analysis the expression of cbfa1 through the RT-PCR method(3 bottles totally every day).Observe the strongest time of expression of Cbfα1 mRNA.(3)The MC3T3-E1 cells were divided into two groups:intermittent exposed to PTH:the cells were cultured with PTH at the first 6h,but without PTH at the subsequent 18h;continuous exposed to PTH:the cells were cultured with PTH in an incubation cycle(24h).Each group was divided into three concentration groups(10^-8mol/L,10^-7mol/ L,10^-6mol/L) and a control group(without PTH in an incubation cycle).the expression of cbfa1mRNA was determined At the time the strongest expression of Cbfα1mRNA in every group(3 bottles in every group).compared on the impact of Cbfα1mRNA expression in the M with different methods,different concentrations of PTH C3T3E1 cell differentiation process.(4)statistical methods:Measurement data was presented as(?),to carry on factorial design analysis of variance,the same group used linear correlation analysis by the statistical analysis software package SPSS 13.0,differences were considered significant at a value of P＜0.05.Results:(1)Semi-quantitative RT-PCR showed that Cbfα1mRNA expression began in the first day of MC3T3-E1 cells induced differentiation process.With MC3T3-E1 cell differentiation,Cbfα1mRNA expression was gradually increased,to the peak in the ninth day.(2)Factorial analysis of variance test results showed that the different stimulation way of hPTH1-34,different concentration,the interaction between the different stimulation way and different concentration influented the Cbfα1mRNA expression(P＜0.05).hPTH1-34 with the concentration of 10^-6mol/L,intermittent stimulation way was strongest to the Cbfα1mRNA expression stimulation.Scatter showed that there was a linear trend between the different concentration and the Cbfα1mRNA expression in the same group. linear correlation analysis showed that there was a positive linear between the two variables in intermittent stimulation group,a negative linear correlation between the two variables in the continuous stimulation group.Conclusions:(1)Cbfα1mRNA expression began in the early period of MC3T3-E1 cells induced differentiation process and expressed continually in MC3T3-E1 cell differentiation process.(2)The expression of Cbfα1mRNA could be up-regulated by intermittent exposed to hPTH1-34,continuous exposed to hPTH1-34 could be reduced Cbfα1mRNA expression, and there was a dose-dependent manner at concentration range of 10^-8-10^-6mol/L.