Studies Of C6 Glioma Cells Apoptosis Induced By Carmustine And Its Mechanism

Objective:To investigate the apoptosis effect of Carmustine on C6 glioma cells and its mechanism in the subcutaneous glioma model in SD rat.Method:C6 glioma cell line was seeded in vitro and C6 glioma cells suspension were implanted into the inguinal subcutaneous regions in 45 SD rats.The culture of 1×10~7/25μl had been successfully implanted in every rats.The survival of rats was observed.The volumes of the growing tumors were periodically measured.When the subcutaneous tumor had been formed,the rats were divided into 2 groups:the controlled group and the treated group.Carmustine was injected into the abdominal cavity to the treated group rats with the same dose.The rats in the treated group were treated in 2 weeks.It was easy to measure the visible tumors with a sliding caliper. Performed the HE dyeing,the immunohistochemistry in order to detect the expression protein of GFAP,Bax and Bcl-2;do RT-PCR to detect the expression diversity of bax/bcl-2 mRNA.Investigating the mechanism of subcutaneous tumor apoptosis induced by Carmustine.Results:4～5 days after the C6 glioma cells were implanted,the subcutaneous tumor in rats could be seen.2 weeks later,there were 32 rats had successfully formed solid tumor in the inguinal subcutaneous region.The successful ratio was 80%.The weight of the tumor in the controlled group were 1.0775±0.20762(g),and in the treated group were 0.7575±0.10181(g).The differences on weight in the two groups was obvious(P＜0.05). The cellular necrosis and apoptosis in the treated goup were more than that in the controlled group.The expression of Bcl-2 mRNA in the treated group was significantly reduced,and the diversity of Bax mRNA in the controlled group was no obvious difference compared to the treated group.The expression of GFAP was masculine in the two groups;and the expression of Bcl-2 protein was dramatically decreased when treated by B CNU(P＜0.01),whereas there were no obvious influence was found to the expression of Bax protein(P＞0.05).Conclusion:carmustine have the proliferation inhibitory effect of C6 glioma.The mechanism of Carmustine in therapy of glioma may be in association with its down-regulating the expression,of Bcl-2 gene and protein,caused augment of the ratio of Bax / Bcl-2.