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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 )增强其对化疗药物顺铂的敏感性
OBJECTIVE: To investigate the inhibitory effect of RibozymeRZ gene against Bcl-2 mRNA on the growth of neuroblastoma (SK-SH) and the chemotherapeutic drug cisplatin. Methods: (1) The defective retroviral vector pDOR-RZ and empty vector pDOR-neo integrated with anti-Bcl-2 mRNA ribozyme were respectively introduced into SK-SH cells by electroporation. (2) The expression of Bcl-2 protein in tumor cells was qualitatively observed by immunocytochemical staining; Bcl-2 protein content was determined by indirect immunofluorescence staining and flow cytometry (FCM). (3) MTT assay chemosensitivity cisplatin. Results: (1) Bcl-2 protein synthesis was inhibited in transfected RZ group, (2) cisplatin sensitization test showed that cisplatin enhanced the killing effect on RZ group. CONCLUSION: Transfection of anti-Bcl-2 ribozyme gene into neuroblasts results in (1) significant inhibition of Bcl-2 protein synthesis and tumor cell death in tumor cells; (2) increased sensitivity to chemotherapeutic agents cisplatin Sex