ABSTRACT
<p><b>OBJECTIVE</b>To detect the transcriptional level of a novel gene F10 associated with the pathogenesis of hydatidiform mole in human cell lines and screen the cell lines with low F10 expression to construct a stable eukaryotic expression system for F10 gene.</p><p><b>METHODS</b>The expression level of F10 mRNA was detected with fluorescent quantitative PCR in A549, 16HBE, Bel7402, HIC, HepG2, 293, PC and MGC cell lines. A549 cell line was transfected with plasmid pRc-CMV2-F10 via electroporation to allow stable F10 expression, and the positive cell clones were selected by G418. The insertion and expression of F10 gene in the A549 cells was analyzed using fluorescent quantitative PCR.</p><p><b>RESULTS</b>F10 mRNA was expressed differentially in these cells lines, and the Bel7402 cells, PC and MGC cells showed the highest F10 mRNA expression, followed by HepG2 and HIC cells and further by 293 cells, and 16HBE and A594 cells had the lowest expression. After transfection, A594 cells showed genomic integration of F10 gene and high expression level of F10 mRNA.</p><p><b>CONCLUSION</b>The pulmonary carcinoma cell line A549 with stable expression of F10 gene has been established, which may facilitate further study of the biological functions of F10 gene.</p>
Subject(s)
Female , Humans , Cell Line, Tumor , Eukaryotic Cells , Metabolism , Gene Expression Regulation, Neoplastic , Genes, Neoplasm , Genetics , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Transfection , Trophoblastic Neoplasms , Genetics , Pathology , Uterine Neoplasms , Genetics , PathologyABSTRACT
<p><b>OBJECTIVE</b>To study the function of F10 gene, a novel hydaditiform mole-related gene.</p><p><b>METHODS</b>A549 cell line was transfected with the F10 gene of forward or reverse sequence or with the empty vector, respectively. The cellular mRNA was extracted after 24 h of transfection to screen for the differentially expressed genes among the 3 transfected and the control cells using differential display-polymerase chain reaction (ddPCR).</p><p><b>RESULTS</b>The bands representing differentially expressed genes were amplified from the cells, and the products were linked to T-Vector for sequence analysis. Several genes were screened by Blasting and their expressions were confirmed by fluorescent quantitative PCR.</p><p><b>CONCLUSION</b>F10 gene is functionally related to cell proliferation and apoptosis.</p>