title: [Construction of subtracted cDNA library in human Jurkat T cell line induced by arsenic trioxide in vitro].
abstract: OBJECTIVE
  To understand the differentially expressed genes in human T lymphocytes induced by arsenic trioxide (As(2)O(3)) and to explore mechanism of its immunotoxicity and immune suppression.
  
  
  METHODS
  Human Jurkat T cell line was treated by arsenic trioxide (5 micromol/L, 24 h) in vitro, as a sample model. Then, the differentially expressed genes were cloned and the subtractive cDNA library from Jurkat T cell line was constructed by suppression subtractive hybridization (SSH). Polymerase chain reaction (PCR) and sequencing techniques were applied to identify positive clones.
  
  
  RESULTS
  The forward subtracted cDNA library contained differentially expressed genes from Jurkat T cell line induced by arsenic trioxide was constructed, including 29 different gene fragments and only replicated one in the subtracted cDNA library identified by PCR and sequencing analysis. These gene sequences were 95%-100% analogous to the genes in public database (GenBank/EMBL). The cDNA library contained oxidative metabolic genes in mitochondria (triose phosphate dehydrogenase, NADH4, pyrophosphate synthase, 16S rRNA ribosome, succinate-CoA ligase and ATP synthase 6); transcriptional and translation genes poly (A) binding protein, t-RNA-guanine transglycoslase, ribosomal protein L23, ribosomal protein S15A, eukaryotic translation initiation factor 3, Rab interaction protein 5, splicing factor-arginine serine rich 5, and ADP-ribosylation factor-like 6 interacting protein), oxide stress related genes (ferritin high chain and high-mobility group protein 2); protein activating and signaling pathway related genes (casein kinase, serine kinase 2 and phosphatidylinositol-four-phosphate adaptor protein-1-associated protein); cell differentiation and apoptosis associated genes (NB4 cell apoptosis related protein and myeloid differentiation primary response protein) and five genes with unknown function (KIAA0092, CGI-147protein, GCI-35, nucleolar phosphoprotein Nopp34 and Mus muscular partial mRNA for hypothetical protein), as well as a novel gene unmatched to the sequence in GenBank.
  
  
  CONCLUSIONS
  The forward subtracted cDNA library contained differentially expressed genes from Jurkat T cell line induced by arsenic trioxide was successfully constructed. And, genes not involved in previous research on arsenic were found. Results of analysis for these genetic function suggested that there should be many genes involved in process of T lymphocytes apoptosis or injury induced by arsenic trioxide and that there should still be many genes associated with arsenic that were not reported in the past.
authors.affiliation: Department of Infectious Diseases, First Affiliated Hospital, Xinjiang Medical University, Urumuqi, Xinjiang 830054, China.
authors.forename: Yue-xin
authors.initials: YX
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: Kai-tai
authors.initials: KT
authors.lastname: Liu
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
authors.forename: Xin-hua
authors.initials: XH
authors.lastname: Chen
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.forename: Guo-quan
authors.initials: GQ
authors.lastname: Wang
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D000077237:Arsenic Trioxide; D001152:Arsenicals / Q000494:pharmacology*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D015723:Gene Library; D006801:Humans; D019169:Jurkat Cells / Q000187:drug effects* / Q000378:metabolism; D009693:Nucleic Acid Hybridization / Q000379:methods; D010087:Oxides / Q000494:pharmacology*; D017422:Sequence Analysis, DNA
chemical_list: D000970:Antineoplastic Agents; D001152:Arsenicals; D018076:DNA, Complementary; D010087:Oxides; D000077237:Arsenic Trioxide