放线菌素D和替尼泊甙对大鼠胶质瘤细胞增殖影响的实验研究

The influences of Actinomycin D and VM-26 on proliferation in rat glioma cell line C_6

目的研究放线菌素D(Actinomycin D,ACTD)和替尼泊甙(Teniposide,VM-26)对大鼠胶质瘤细胞增殖的影响。方法应用免疫细胞化学方法,观察C6细胞经不同剂量ACTD和VM-26作用48h后PCNA、CyclinD1表达的改变,FCM法分析细胞凋亡;流式细胞术分析细胞周期变化,3H-TdR掺入法分析细胞增殖。结果对照组PCNA和CyclinD1均为强阳性;经放线菌素与VM-26作用后,PCNA和CyclinD1表达均受到显著的抑制,尤以放线菌素D组效果最佳,与对照组比较差别显著,FCM法发现C6组几乎没有凋亡细胞,高浓度的ACTD和VM-26组的凋亡率明显增加(χ2=63.714,P<0.01);流式细胞分析发现ACTD和VM-26组S期指数较对照细胞明显减少,3H-TdR掺入法发现与C6组比较,ACTD和VM-26组48h存活率均明显下降,且两者之间存活率也有明显差异(χ2=52.223,P<0.01);以ACTD组下降最显著。结论放线菌素D可显著抑制胶质瘤细胞恶性增殖并诱导细胞凋亡,可以成为胶质瘤化疗的一种新策略,值得进一步进行体内实验。

Objective To study the effects of actinomycin D （ACTD） and VM-26 on proliferation in rat glioma ceils C6. Methods The inhibition effect of actinomycin D or VM-26 was compared in treatment of rat glioma cell line by 3H-TdR method in vitro. The cell apoptosis and cell cycle were analysed by flow cytometry. The expression of PCNA and cyclin D1 was observed by immunohistochemistry. Results There were nearly no apoptotic cells found in parental cells as indicated by FCM assay, however, apoptosis was increased in cells treated by high level ACTD or by VM-26 （x^2 = 63. 714 ,P 〈 0.01 ）. The flow cytometric analysis showed that the S phase fraction （SPF） was lowered in ACTD treated cells than that in parental cells. Compared to parental cells, the 3 H-TdR method assay indicated the survival rates of treated cells dramatically dropped down trerated by treated with ACTD or VM-26 after 48hours late. The ACTD treated cells demonstrated much lower survival rate than VM-26 treated cells （x^2 = 52. 223, P 〈 0.01 ）. Meanwhile, protein levels of PCNA and CyclinD1 decreased sginificantely after treated in C6 cells compared to C6 cells, still much lower in ACTD treated groups than that in VM-26. Conclusions Compared with the two drags, ACTD could more strongely inhibit glioma cell growth than VM-26. The results demonstrate that ACTD can significantly inhibit proliferation, decrease the degree of malignancy. Whereas the study in vivo should be explored in the future.