Study On Anti-glioma Effect Of Radiation By Per1,Per2 Genes In Vitro

Objective: To explore the per1,per2 regulate on C6 glioma cells and NIH3T3 cell on radiation in the different time, to research the effect of Per1,Per2 on the C6 glioma cells and NIH3T3 cells on the apoptosis rate and effects of proliferation capacity by radiation.Methods:The culture C6,NIH3T3 cells stimulated by PMA,the expression of per1,per2 at the differented times was examined by Real-time PCR.To Investigate the rate of apoptosis of C6,NIH3T3 cell at differented expression of per1,per2 by Annexin-V/PI.The pcDNA3.1(+)per1,pcDNA3.1(+)per2 were transfected into the C6 and NIH3T3 cell, They were irradiated with X-ray and to investigate the rate of apoptosis and to investigate the capbilty of proliferation.Rseults: Following the treatment with PMA ,the per2 gene of C6 cell displayed a circadian rhythm which then rapidly decreased to basal levels at 16h ,followed by a second peak and trough at approximately 28h and 40h. However,PMA did not seem to affect rPer1 mRNA expression, in contrast to the acute induction and the subsequent repression of rper2.Rper1,in C6 cell,was continuously upregulated for 48h after exposure to PMA.For NIH3T3 cell, after the transient exposure to PMA expression levels of mper1 and mper2 mRNAs oscillated with an approximate period length of 24h which rapidly decreased to basal levels at 12h, followed by a second peak and trough at approximately 24 and 36h in NIH3T3 cell.High expression of rPer2 led to Cell Cycle Arrest at G2/M phase in C6 cells. At 28h,the peak level of rper2,the rate of C6 cell were significantly higher than the cells of low level at 16h. For NIH3T3,apoptosis rate of cells at 1h and 24h were significantly decreased Compared with 12h and control group.At the time, It was found that the expression of rPer2 inhibit the gowth of C6 cells.At 28h after treatment ,proliferation index(PI) of cells was clearly lower than control group and 16 h.It also show no differentiation into rper1 over expression and 28 h,the proliferation index of rper2 over expression was lowest.Conclusion: PMA can induce the circadian oscillation of Per2 in C6 cells.The Per1,Per2 gene protective effect to NIH3T3 when rerrived the X-ray,and Per2 could decrease the rate of apoptosis in C6 cell.