The effect of fragile histidine triad gene on the apoptosis of cholangiocarcinoma cells

Abstract

Objective To discuss the effect of fragile histidine triad (FHIT) gene on the apoptosis of QBC939 cells and its modulatory effects on Cyclin DI. Methods The recombinant eukaryotic expression plasmid was constructed. The result was checked by DNA testing. Then the FHIT/pcDNA3.1 was transfected into QBCV39 cells. The apoptosis was detected by flow cytometer (FCM), and the data was analyzed by chi-square test. The expression change of Cyclin D1 mRNA and protein was detected by real-time reverse tran-scription polyroerase chain reaction (RT-PCR) and Western blotting respectively. All the experimental subjects were divided into three groups: QBC939 was cultured in natural status (nature control group) ; QBC939 was transfected with blank plasmid (blank control group) and QBC939 was transfected with FHIT/pcDNA 3.1 (experimental group). Results The human FHIT recombinant eukaryotic expression plasmid was constructed and transfected successfully. The apoptosis rate after transfection was significantly promoted (25.8% vs 30.9% vs 55.4% ,P<0.05). The expression of Cyclin D1 mRNA was decreased from the original cell line of 0.03-fold. And that of protein was decreased too (0.33±0.02 vs 0.33±0.01 vs 0.14±0.02,P<0.05). Conclusion The human FHIT recombinant eukaryotic expression plasmid provides molecular biology tool for further funda-mental and clinical research of cholangiocarcinoma. FHIT gene can promote apoptesis of QBC939 cells and re-duce the expression level of Cyclin D1. Key words: Cholangiocarcinoma; Transfection; Apoptosis; Cyclin D1