title: Transforming growth factor beta1 gene variation Leu10Pro affects secretion and function in hepatic cells.
abstract: BACKGROUND
  Our previous work revealed transforming growth factor beta1 (TGFβ1) gene polymorphisms are associated with susceptibility to hepatocellular carcinoma and liver cirrhosis. However, no further study of functional substitution in hepatic cells has yet been reported.
  
  
  OBJECTIVE
  This study was designed to uncover the functional mechanisms of TGFβ1 gene polymorphisms in the pathogenesis of liver diseases.
  
  
  METHODS
  Two recombinant TGFβ1 expression plasmids containing TGFβ1 codon 10 Leu/Pro variation were constructed with CMV promoter and transfected into human hepatoma cell lines (HepG2 and SMMU 7721), hepatic stellate cells (LX-2), and immortalized hepatocytes (L02). The secretion capacities of TGFβ1 protein in the transfected cells were determined by ELISA. Apoptosis, proliferative activity, and expression of CD 105, CD83, and CD80 were also measured by use of flow cytometry.
  
  
  RESULTS
  The ELISA results showed that cells transfected with CMV-Pro10 were more capable of TGFβ1 secretion than those transfected with CMV-Leu10. Functionally, CMV-Pro10 was more apoptosis-protective and induced more proliferation than CMV-Leu10 in transfected hepatic cells. Pro10 up-regulated expression of CD105 and down-regulated expression of CD83.
  
  
  CONCLUSIONS
  TGFβ1 gene Leu10Pro variation in signal peptide has significant effects on TGFβ1 secretion and functions in hepatic cells.
authors.affiliation: Department of Laboratory Medicine, Eastern Hepatobiliary Hospital, Second Military Medical University, 225 Changhai Road, Shanghai 200438, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Gu
authors.forename: Xin
authors.initials: X
authors.lastname: Ji
authors.forename: Le-Hua
authors.initials: LH
authors.lastname: Shi
authors.forename: Chang-Hong
authors.initials: CH
authors.lastname: Yi
authors.forename: Yun-Peng
authors.initials: YP
authors.lastname: Zhao
authors.forename: Ai-Hua
authors.initials: AH
authors.lastname: Wang
authors.forename: Lun-Gen
authors.initials: LG
authors.lastname: Lu
authors.forename: Wen-Bo
authors.initials: WB
authors.lastname: Yu
authors.forename: Chun-Fang
authors.initials: CF
authors.lastname: Gao
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D017209:Apoptosis; D002460:Cell Line; D045744:Cell Line, Tumor; D003062:Codon; D017931:DNA Primers; D015536:Down-Regulation; D000071063:Endoglin; D004797:Enzyme-Linked Immunosorbent Assay; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D010957:Plasmids; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic; D011956:Receptors, Cell Surface / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000235:genetics* / Q000378:metabolism; D015854:Up-Regulation; D000097802:CD83 Antigen
chemical_list: D015703:Antigens, CD; D003062:Codon; D017931:DNA Primers; D000071063:Endoglin; D007136:Immunoglobulins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D053773:Transforming Growth Factor beta1; D000097802:CD83 Antigen; C064206:ENG protein, human