中国免疫学杂志2017，Vol.33Issue(2)：196-200,205,6.DOI:10.3969/j.issn.1000-484X.2017.02.007
重组Ser17突变型 hIFN-β腺病毒的构建及其对结肠癌细胞的增殖抑制作用
Inhibited effects of a recombinant adenovirus expressing Ser17 mutant human in-terferon-βon proliferation of a human colon carcinoma cell line in vitro
摘要
Abstract
Objective:To investigate the effect of replication-defective recombinant adenovirus expressing Ser17 mutant human IFN-β on the cell proliferation and cycle regulation of a human colon carcinoma cell line in vitro. Methods:Ser17 mutant human IFN-βgene was inserted into a shuttle plasmid ( pshuttle-CMV ) . The pshuttle-Ser17 hIFNβ was transformed into Ecoli BJ5183 to make a homologous recombination with adenovirus skeleton genome (pAdeasy). And the recombinant plasmid pAd-Ser17hIFNβwas transfected into HEK-293 cell to produce mature adenovirus particles. The change of proliferation and cell cycle of rAd infected HT-29 were analyzed by MTT assay and FACS, respectively. Results: The recombinant adenovirus ( rAd-Ser17 hIFNβ) with the titer of 109. 125 CCID50/ml was produced. HT29 cells could be infected with the rAd-Ser17 hIFNβin 20 MOI. The expression of Ser17 hIFN-βin HT-29 could be detected by Western blot. Significantly, the rAd-Ser17 IFNβ inhibited the growth of HT-29 cells and prevented cell cycle progression in S phage in vitro. Conclusion:The recombinant adenovirus expressing Ser17-mutant human IFN-βcould suppress the pro-liferation of colon carcinoma cell in vitro. It provides an experimental basis to apply the IFN-βgene therapy for colon carcinoma in the future.关键词
Ser17 hIFN-β/结肠癌/重组腺病毒Key words
Ser17 hIFN-β/Colon carcinoma/Recombinant adenovirus分类
医药卫生引用本文复制引用
唐丽媛,谢力,李楠,缪希松,王元,芮丹云,童毅,王波,李臣,句红萍..重组Ser17突变型 hIFN-β腺病毒的构建及其对结肠癌细胞的增殖抑制作用[J].中国免疫学杂志,2017,33(2):196-200,205,6.基金项目
本文受昆明市科技计划重点项目(2015-1-S-00877)、昆明学院人才项目(YJ216002)和昆明学院科学研究项目资助(XJL12026)。 （2015-1-S-00877）
