title: [Construction of adenovirus vector expressing IL-24 and E1A and its inhibition of SMMC-7721].
abstract: We constructed the recombinant adenovirus vector expressing IL-24 and E1A (Ad-IL-24-E1A) and investigated the inhibition of Ad-IL-24-E1A on SMMC-7721 hepatocellular carcinoma in vitro. We amplified IL-24 gene by PCR using pAdTrack-IL-24 as template. The IL-24 gene was cloned into pAdTrack-IRES at the Bgl II and Sal I site to form pAdTrack-IL-24-IRES. E1A digested from pAdTrack-E1A was cloned into the pAdTrack-IL-24-IRES at the Xho I and EcoR V site to form the pAdTrack-IL-24-IRES-E1A. We co-transformed both pAdTrack-IL-24-IRES-E1A and pAdeasy-1 digested by Pme I and packaged to obtain Ad-IL-24-E1A. Ad-IL-24-E1A at 50 MOI infected SMMC-7721 cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay determined cell proliferation. Flow cytometry detected Cell apoptosis. The apoptotic rate of SMMC-7721 cells was 52% 48 h after infection with Ad-IL-24-E1A. The result showed that the growth of SMMC-7721 cells was significantly inhibited by Ad-IL-24-E1A at the MOI of 50.
authors.affiliation: No. 1 Affiliated Hospital of Soochow University, Suzhou 215001, China.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Wang
authors.forename: Jingcheng
authors.initials: J
authors.lastname: Miao
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Xie
authors.forename: Weihua
authors.initials: W
authors.lastname: Sheng
authors.forename: Yunbo
authors.initials: Y
authors.lastname: Shan
authors.forename: Jicheng
authors.initials: J
authors.lastname: Yang
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D017500:Adenovirus E1A Proteins / Q000096:biosynthesis / Q000235:genetics*; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007378:Interleukins / Q000096:biosynthesis / Q000235:genetics*; D008113:Liver Neoplasms / Q000473:pathology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D000099180:Interleukin-24
chemical_list: D017500:Adenovirus E1A Proteins; D000970:Antineoplastic Agents; D007378:Interleukins; D011994:Recombinant Proteins; D000099180:Interleukin-24