[Effect of recombinant plasmid of Helicobacter pylori ureB gene on gastric epithelial cell].
To investigate the effect of recombinant plasmid of Helicobacter pylori ureB gene on gastric epithelial cell. The full length sequence of ureB gene from NCTC11637 was amplified by PCR. The recombinant plasmid was constructed by cloning the open reading frame (ORF) of ureB into the eukaryotic expression vector pcDNA3.1, and was transfected SGC-7901 cells, then the clones resisting Hygromycin were screened. mRNA expression of ureB of transfected cells was detected by RT-PCR. The effect of recombinant plasmid of Hp ureB gene on cell phenotype was observed by fluorescence strain, on proliferation by MTT, on apoptosis and cell cycles by flow cytometry, respectively. The positive clones of ureB (SureB) appeared cell membrane budding and cell shrinkage. MTT assay showed there was no statistic significance between the SureB and SpcDNA3.1 which were transfected only by pcDNA3.1 (P > 0.05), suggesting that the growth of SureB were not inhibited. The apoptosis rate of SureB was higher than that of SpcDNA3.1 (P = 0.007). Analysis for cell cycle showed that in SureB cells the proportion of S phase increased, the proportion of both G2/M and G0/G1 phase decreased. Positive transfection of ureB gene into SGC-7901 can change cell phenotype and induce cell apoptosis.
Zhang
Jing
J
Fu Jian Medical University, Department of Etiological Biology, Fuzhou 350004, China. zj-78@sohu.com
She
Fei-fei
FF
Chen
Yue-xiu
YX
Chen
Hao
H
D017209:Apoptosis; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004847:Epithelial Cells / Q000166:cytology; D005434:Flow Cytometry; D005753:Gastric Mucosa / Q000166:cytology*; D016480:Helicobacter pylori / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D010641:Phenotype; D010957:Plasmids; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014510:Urease / Q000235:genetics* / Q000378:metabolism
D014510:Urease