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Objective To investigate the effect of expression of pituitary tumor transforming gene(PTTG) on the expression of inhibitory of vascular endothelial growth factor(VEGF).Methods The constructed(recomibinant) vector,pcDNA3.1-PTTGas that contained full-length antisense PTTG,was transfected into the cholangiocarcinoma cell line QBC939 in different quantity.After 48h incubation,the variation of expressions of PTTG and VEGF mRNAs and proteins were observed by RT-PCP and Western-blot method.Results(After) successful transfecting the recomibinant vector into QBC939,the mRNAs and proteins of PTTG and VEGF were all inhibited.Linear correlation analysis showed positive correlation between descendent extents of PTTG and VEGF(P
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Objective To investigate the effct of PTTG gene on the growth and sensitivity to 5-FU of cholangiocarcinoma cell.Methods The pcDNA3.1-PTTGas that contained full-length antisense PTTG,and pcDNA3.1(+) was transfected into the cholangiocarcinoma cell line QBC939.We successfully established and identified cell sublines tQBC939(PTTG-),tQBC939(pcDNA3.1).Cell growth curve was mapped out,cell proliferation rate was assessed by MTT assay and cell cycle percentage was counted by flow cytometry.After treatment with 5-FU,cell survival rate was assessed by MTT assay and IC50 was calculated.Flow cytometry and Hoechst staining were used to demonstrate discrepancy of cell apoptosis rates.Results Compared with other two groups,tQBC939(PTTG-) grew more rapidly and had a higher proportion of cells in S phase,whereas less cells in G2/M phase(P
ABSTRACT
Objective:To investigate the different killing effects on human hilar cholangiocarinoma cells FRH with cytosine deaminase(CD) and herpes simplex virus thymidine kinase(HSV tk)double suicide genes coexpression compared with single gene mediated by retrovirus.To find a more efficient and low toxicity suicide gene therapy for hilar cholangiocarinoma.Methods:CD and HSV tk double suicide genes were transfected into PA317 cells using lipofectamine.The positive clones were picked out and cultured after G418 selected.The viral supernatant was collected.The FRH cells were infected with the virus containing the double suicide genes.After G418 selection,RT PCR was resorted to demonstrated the successful transcription of CD and HSV tk genes.The FRH/CD+tk and FRH cells in culture were respectively treated with 5 Fc and /or GCV.The cytoxicity efficacy was evaluated by microculture tetrajolium test (MTT) method.Results:The virus containing double suicide gene was produced in PA317 cells.Double suicide genes were stably expressed in RFH cells after being infected with the virus.The killing effect of combination 5 Fc with GCV on FRH/CD+tk cells is more effective than that of using 5 Fc or GCV alone.Conclusion:The CD+TK/5 Fc+GCV co expression system is more effective for killing effects on FRH cells than that of CD/5 Fc or tk/GCV system alone.