TRAIL和ACNU对实验性大鼠脑胶质瘤的治疗作用

Therapeutical Effect of TRAIL and ACNU on Glioma in Experimental Rat

[目的]观察TRAIL和嘧啶亚硝脲(ACNU)对实验性大鼠脑胶质瘤的治疗作用,并探讨TRAIL和ACNU联合在诱导细胞凋亡中的作用机制。[方法]制备Wistar大鼠C6细胞胶质瘤动物模型,并随机分成4组:TRAIL+ACNU治疗组(A组)、ACNU治疗组(B组)、TRAIL治疗组(C组)、生理盐水对照组(D组)。观察动物的生存状况及肿瘤大体情况,测量肿瘤的体积、计算肿瘤的抑制率;观察TRAILR2/DR5免疫组织化学表达变化情况;电镜观察肿瘤细胞变化;流式细胞仪检测肿瘤细胞凋亡情况。[结果]A组与B组的大鼠生存较好,而C组与D组的大鼠生存差。肿瘤体积分别为A组(19.00±2.59)mm3、B组(68.76±5.56)mm3、C组(230.31±13.94)mm3、D组(238.84±10.64)mm3。除C组和D组相比无显著差异外,4组间两两比较均有显著差异(P<0.05)。电子显微镜观察,A组和B组可见明显凋亡小体,C组与D组少见。肿瘤细胞凋亡率A组(20.38%±1.62%)大于B组(14.85%±2.41%),差异有显著性(P<0.05);C组(0.44%±0.21%)与D组(0.35%±0.24%)相比较差异无显著性(P>0.05)。[结论]ACNU引起肿瘤细胞凋亡,并可诱导大鼠脑胶质瘤细胞表达DR5,使其对TRAIL诱导的凋亡敏感,TRAIL和ACNU体内联合应用具有协同作用。

[Purpose] To investigate the therapeutical effect of TRAIL and ACNU on glioma in experimental rat,and to explore the mechanism of their cooperative effect on induced tumor-cell apoptosis. [Methods] Glioma animal model was established by injecting C6 cells into the right caudate putamen of Wistar rats. The rats were randomly divided into 4 groups：TRAIL＋ACNU group（group A）,ACNU group（group B）,TRAIL group（group C） and saline control group（group D）. Survival and tumor gross condition were observed. Tumor volume and inhibition ratio of tumor cells were determined. Expression of TRAIL R2/DR5 with immunohistochemistry,alteration under electron microscope and apoptosis of tumor cells were observed. [Results] The survival in A or B groups was better than that in C or D group. The tumor volume of group A was （19.00±2.59）mm3; group B,（68.76±5.56）mm3; group C,（230.31±13.94）mm3; and group D,（238.84±10.64）mm3. Significant differences were found between the 4 groups （P0.05）,except between group C and D. Apoptotic bodies in A and B groups were obvious electronmicroscopically,and less in group C and D. The apoptosis rate of group A （20.38%±1.62%） was higher than that of group B （14.85%± 2.41%）,with significant difference（P0.05）. No significant difference of apoptosis rate was found between group C（0.44%±0.21%） and group D（0.35%±0.24%） （P0.05）. [Conclusions] ACNU causes tumor cells apoptosis and induces DR5 expression in glioma cells,which was sensitive to apoptosis that induced by TRAIL. Combination of TRAIL and ACNU in vivo has synergic effect to induce cell apoptosis.