摘要
目的 :研究抗Bcl- 2mRNA的锤头型核酶 (RibozymeRZ)基因抑制神经母细胞瘤 (SK -N -SH)生长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Bcl- 2mRNA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 )用免疫细胞染色法定性观察肿瘤细胞内Bcl- 2蛋白的表达 ;用间接免疫荧光染色法和流式细胞仪 (FCM)测定Bcl- 2蛋白含量。 (3)MTT法检测化疗药物顺铂敏感性。结果 :(1)转染RZ组Bcl- 2蛋白合成受到抑制 ,(2 )顺铂增敏试验表明顺铂对转RZ组杀伤作用增强。结论 :将抗Bcl- 2核酶基因转入神经母细胞内 ,(1)可明显抑制肿瘤细胞内Bcl- 2蛋白的合成 ,促进瘤细胞死亡 ;(2 )
Objective: To observe whether tumor cell can be inhibited and the sensitivity of SK-N-SH cell transferred to cisplatin can be increased through transducting a ribozyme against Bcl- 2 into neuroblastoma cell line (SH-N-SH).Methods: pDOR-RZ retroviral vector was transduced into neuroblastoma cell by electroporation. By immunocytochemistry 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. Conclusion:The ribozyme gene against Bcl-2 can inhibit the synthesis of Bcl-2 protein and tumor growth , and increase the sensitivity of chemotherapeutic agent.
出处
《重庆医科大学学报》
CAS
CSCD
2000年第2期123-124,127,共3页
Journal of Chongqing Medical University
