[1]尹昌林,吕胜青,黄轶,等.pSiRNA-Notch1逆转录病毒载体的构建及对恶性脑胶质瘤细胞的作用[J].第三军医大学学报,2007,29(12):1211-1214.
 YIN Chang-lin,LU Sheng-qing,HUANG Yi,et al.Construction of pSiRNA-Notch1 recombinant retroviral vector and its effect on malignant glioma cells in vitro[J].J Third Mil Med Univ,2007,29(12):1211-1214.
点击复制

pSiRNA-Notch1逆转录病毒载体的构建及对恶性脑胶质瘤细胞的作用(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第12期
页码:
1211-1214
栏目:
论著
出版日期:
2007-06-30

文章信息/Info

Title:
Construction of pSiRNA-Notch1 recombinant retroviral vector and its effect on malignant glioma cells in vitro
作者:
尹昌林吕胜青黄轶李光辉唐莉杨辉
第三军医大学:新桥医院神经外科,新桥医院全军肿瘤诊治中心,重庆市肿瘤生物技术研究所,西南医院乳腺疾病中心;重庆医科大学生物化学与分子生物学教研室
Author(s):
YIN Chang-lin LU Sheng-qing HUANG Yi LI Guang-hui TANG Li YANG Hui
Department of Neurosurgery, Institute for Cancer Research, Xinqiao Hospital, Chongqing 400037; Breast Disease Center, Southwest Hospital, Third Military Medical University Chongqing 400038; Department of Biochemistry and Molecular Biology, Chongqing Medical University, Chongqing 400010, China
关键词:
pSiRNA-Notch1逆转录病毒载体恶性脑胶质瘤基因治疗
Keywords:
pSiRNA-Notch1 retroviral vector malignant glioma gene therapy
分类号:
R394-33;R73-362;R739.41
文献标志码:
A
摘要:
目的    建立逆转录病毒介导的人Notch1基因RNA干扰体外表达体系并观察对恶性脑胶质瘤细胞的作用。    方法    将人Notch1基因RNAi双链转录DNA片段重组到逆转录病毒质粒Psilencer 5.1-H1 Retro中,构建成携带人Notch1基因RNAi逆转录病毒载体pSiRNA-Notch1,经PT67细胞包装后,产生的重组逆转录病毒感染U251和CHG-5恶性脑胶质瘤细胞,用WST-8、RT-PCR和Western blot分别检测对转染细胞活性、人Notch1 mRNA和蛋白表达的影响。    结果    重组pSiRNA-Notch1质粒经测序鉴定正确。重组逆转录病毒滴度可达224×104cfu/ml,感染U251和CHG-5恶细胞后3 d能明显抑制细胞生长,RT-PCR和Western blot检测人Notch1 mRNA和蛋白表达水平明显低于阴性对照组和未干扰组。    结论    携带人Notch1基因RNAi双链转录DNA片段的逆转录病毒有明显的抑制恶性脑胶质瘤细胞生长作用,为下一步开展基因治疗恶性脑胶质瘤奠定了基础。
Abstract:
Objective    To construct a retrovirus-mediated expression system containing double strands DNA for RNA interference on human Notch1 gene and study its inhibitory effect on human glioma cell lines U251 and CHG-5 in vitro.      Methods    A recombinant retroviral vector pSiRNA-Notch1 was generated by cloning a double strands DNA for RNA interference on human Notch1 gene into a retroviral vector Psilencer 5.1-H1 Retro. Human glioma cell lines U251 and CHG-5 were infected with the viral supernatant from the PT67 clones. After 3 d, the viability, Notch1 mRNA and protein of the transfected cells were examined by WST-8 assay, RT-PCR and Western blot.     Results    The pSiRNA-Notch1 recombinant retroviral vector had been constructed correctly. The titer assayed on NIH3T3 cells was up to 224×104 cfu/ml. Three days after transfection, the viability, Notch1 mRNA and protein of the transfected cells decreased significantly.     Conclusion    The constructed pSiRNA-Notch1 retroviral vector shows effective inhibition effect on the viability in human malignant glioma cells, with potential utility in the gene therapy for human malignant glioma.

相似文献/References:

[1]樊锐太,胡勇,刘俊启,等.硼替佐米对U87胶质瘤放射增敏及细胞周期分布的影响[J].第三军医大学学报,2012,34(08):758.
 Fan Ruitai,Hu Yong,Liu Junqi,et al.Effect of bortezomib on radiosensitivity of U87 glioma and distribution of cell cycle[J].J Third Mil Med Univ,2012,34(12):758.
[2]姜德福,邹咏文,陈广鑫,等.大剂量化疗、大剂量类固醇并用自体骨髓移植治疗恶性脑胶质瘤(附2例报告及文献复习)[J].第三军医大学学报,1989,11(03):0.[doi:10.16016/j.1000-5404.1989.03.017 ]

更新日期/Last Update: 2008-10-23