title: Transfection of thymidine phosphorylase cDNA to human hepatocellular carcinoma cells enhances sensitivity to fluoropyrimidine but augments endothelial cell migration.
abstract: OBJECTIVE
  To investigate the effects on sensitivity to fluoropyrimidine and endothelial cell (EC) migration by transfection with thymidine phosphorylase (TP) cDNA to a hepatocellular carcinoma (HCC) cell line SMMC-7721.
  
  
  METHODS
  SMMC-7721 was transfected with pcDNA3.1/zeo (+) with human TP cDNA. TP mRNA expression was determined by RT-PCR. Sensitivity to fluoropyrimidine was determined by 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazolium bromide (MTT) assay. Induction of EC migration was detected by Boyden chamber assay.
  
  
  RESULTS
  The construction of pcDNA3.1/zeo(+)-TP was verified by digestion with restriction endonuclease Apa1. When comparison was made between SMMC-7721 cell clone transfected with pcDNA3.1/zeo(+)-TP (S-TP) and control clone transfected with pcDNA3.1/zeo(+) (S-vector), we found that TP mRNA expression level was much higher in S-TP, being 2.09+/-0.16 vs 0.48+/-0.06 in S-vector (P < 0.01), sensitivity to 5'-deoxy-5-fluorouridine (5'-dFUrd, a prodrug of 5-fluorouracil) in S-TP was significantly enhanced compared with that in S-vector (IC(50); 56.81+/-9.85 micromol/l vs 162.25+/-11.03 micromol/l, P < 0.01), and the culture medium of S-TP possessed more potential to induce EC migration than that of S-vector (the number of ECs appearing on the outer surfaces of the membrane was 275+/-29 vs 122+/-35 per field, P < 0.01).
  
  
  CONCLUSIONS
  Sensitivity to 5'-dFUrd could be enhanced by transfection with TP cDNA for SMMC-7721 cells. However, EC migration was also promoted at the same time. Therefore, transfection with TP alone might have no potential to enhance anti-tumoral effects of fluoropyrimidine in HCC.
authors.affiliation: Liver Cancer Institute and Zhongshan Hospital, Fudan University, Shanghai, 200032, People's Republic of China.
authors.forename: Jian
authors.initials: J
authors.lastname: Zhou
authors.forename: Yong-Sheng
authors.initials: YS
authors.lastname: Xiao
authors.forename: Zhao-You
authors.initials: ZY
authors.lastname: Tang
authors.forename: Jia
authors.initials: J
authors.lastname: Fan
authors.forename: Zhi-Quan
authors.initials: ZQ
authors.lastname: Wu
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhao
authors.forename: Qiong
authors.initials: Q
authors.lastname: Xue
authors.forename: Zao-Zhuo
authors.initials: ZZ
authors.lastname: Shen
authors.forename: Yin-Kun
authors.initials: YK
authors.lastname: Liu
authors.forename: Sheng-Long
authors.initials: SL
authors.lastname: Ye
mesh_terms: D000951:Antigens, Neoplasm / Q000032:analysis; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D002465:Cell Movement / Q000187:drug effects; D004250:DNA Topoisomerases, Type II / Q000032:analysis; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000032:analysis; D018572:Disease-Free Survival; D005467:Floxuridine / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016019:Survival Analysis; D013939:Thymidine Phosphorylase / Q000235:genetics*; D014162:Transfection
chemical_list: D000951:Antigens, Neoplasm; D000964:Antimetabolites, Antineoplastic; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D005467:Floxuridine; D013939:Thymidine Phosphorylase; D004250:DNA Topoisomerases, Type II; C025034:doxifluridine