核酶诱导神经母细胞瘤凋亡的实验研究

目的 研究抗Bcl-2mRNA的锤头型核酶（RibozymeRZ)基因诱导神经母细胞瘤（SK－N－SH）凋亡以及增强化疗药物顺铂的敏感性，方法（1）应用电穿孔法，分别将整合有抗Bcl02mRNA核酶的缺陷性逆转录病毒载体pDOR-RZ和空载体pDOR-neo导入SK－N－SH细胞，（2）用免疫组化和间接免疫荧光染色法观察Bcl-2蛋白表达。（3）用流式细胞仪（FCM）检测细胞调率及细胞周期的变化。（4）电镜观察转染细胞形态变化。结果 （1）转染RZ组Bcl-2蛋白合成受到抑制，（2）细胞周期显示，转染RZ组可见凋亡峰，发生Go/G1期阻滞，电镜观察呈典型的凋亡形态改变。（3）顺铂增敏试验表明顺铂对转RZ组杀伤作用增强。结论 逆转录病毒介导的抗Bcl-2核酶基因可抑制肿瘤细胞内Bcl-2蛋白的合成，促进瘤细胞凋亡，并可增强其对化疗药物顺铂的敏感性。

The apoptosis of neurolastoma cell induced by ribozyme

Objective Through transducting a ribozyme against Bcl 2 into neuroblastoma cell line (SH N SH), to observe whether tumor cell can be inhibited and the sensitivity of SK N SH cell transferred to cisplatin can be increased. Methods pDOR RZ retroviral vertor was transducted into neuroblastoma cell by electroporation. By immuocytochemisty and indirect immunofluorescence dyeing methods, the changes of Bcl 2 expression were observed; and the sensitivity of cisplatin to transferred cell was quantified by a modified 2, 5 diphenyl tetrazolium bromide (MTT) colorimetric assay. Results (1) The synthesis of BCL 2 protein was inhibited; (2) the sensitivity of the cell with RZ to cisplatin was increased. Conclusions The ribozyme gene against Bcl 2 can inhibite the synthesis of Bcl 2 protein and tumor growth, increase the sensitivity of hemotherapeutant.