逆转录病毒载体介导的反义c-myc抗白血病作用的研究  

Effects of anti-leukemia by antisense c-myc by retrovirus vector

作　　者：赵新永[1] 殷莲华[1] 孔宪寿[1] 杨轶群[1] 

机构地区：[1]上海医科大学病理生理教研室,上海200032

出　　处：《中国病理生理杂志》2001年第11期1043-1047,共5页Chinese Journal of Pathophysiology

基　　金：上海市教委高教科技发展基金资助项目 (No .97JG0 5 0 5 3)

摘　　要：目的 :研究c-myc基因在L6 5 6 5小鼠白血病发生中的作用以及抑制其表达所起的治疗作用。方法 :用逆转录病毒载体pGNC构建一个含有小鼠反义c -myc基因的逆转录病毒表达系统pGNCas。用磷酸钙沉淀法将pGNCas导入包装细胞PA317,筛选获得病毒生产细胞。用pGNCas病毒感染L6 5 6 5白血病克隆细胞 ,筛选后 ,获得抗G418阳性克隆细胞L6 5 6 5as。PCR方法检测反义c -myc序列是否稳定整合入L6 5 6 5as细胞中。最后从细胞生长、形态、周期、软琼脂集落形成能力以及c -myc基因表达方面观察其恶性和表型的改变。结果 :L6 5 6 5as细胞形态变圆 ,大小较一致。生长曲线显示 :L6 5 6 5as细胞生长被抑制 ;流式细胞仪检测细胞周期 :L6 5 6 5as阻滞于G0 /G1期 ,S期细胞减少 ;软琼脂集落形成试验发现L6 5 6 5as细胞形成克隆数明显减少。免疫组化检测 :L6 5 6 5as细胞c-myc蛋白表达呈弱阳性 ,而对照组细胞呈强阳性。结论 :c -myc基因在L6 5 6 5小鼠白血病的发生中起着重要的作用 ;抑制c-myc的表达 ,可部分逆转L6 5 6 5白血病的恶性表型和恶性行为。AIM: To study the role of c-myc oncogene in L6565 leukemia oncogenesis and the effects of therapy by inhibition of its expression with antisense c-myc. METHODS: A recombinant retroviral vector containing antisense c-myc of the murine (pGNCas)was constructed and then transfected into PA317 cells by the method of calcium phosphate precipitation. L6565 clone cells were infected with retrovirus particles. Stable integretion of antisense c-myc was shown by PCR. The change of the malignance and phenotype of L6565as were detected by the examination of the growth, morphology, cells cycle, agar assay and expression of c-myc. RESULTS: The shape of most L6565as cells became spherical. The growth of L6565as was inhibited compared to control cells. The analysis of cells cycle: L6565as cells were arrest in G 0/G 1 phase, decreased in S phase. The ability of L6565as cells to form colony in soft agarose was significantly suppressed. c-myc in L6565as cells was lowly expressed. CONCLUSION: (1)c-myc plays a critical role in L6565 leukemia oncogenesis; (2)Inhibition of expression of c-myc makes partly reversion of malignant phenotype of L6565 murine leukemia clone cells.

关 键 词：逆转录病毒科 癌基因 反义寡核苷核类 白血病 C-MYC 

分 类 号：R733.7[医药卫生—肿瘤]