野生型P73β转染对胆管癌细胞的生物学影响

Biological effects of P73β gene on human cholangiocarcinoma cell line QBC939 behavior in vitro

目的应用野生型P73β转染胆管癌QBC939细胞,探讨P73β基因转染对胆管癌细胞生长的抑制机制。方法应用野生型P73β转染胆管癌QBC939细胞,并以未作处理的QBC939细胞作为空白对照。用RT-PCR及Western blotting检测细胞中P21、Cyclin D1及Cdk2表达量的变化。使用四甲基偶氮唑盐法(MTT)和流式细胞仪检测细胞增殖及细胞周期的变化。结果转染了野生型P73β重组质粒的胆管癌QBC939细胞,P21相对表达量增高,Cdk2相对表达量明显减少,胆管癌细胞增殖率受到抑制。结论野生型P73β基因转染后对胆管癌细胞的生长有抑制作用,P73β基因可通过增加P21基因的表达,抑制Cdk2基因的表达,进而作用于S期的细胞,使细胞周期停滞,从而抑制胆管癌细胞增殖。

【Objective】 To investigate the effects of P73 on the cell cycle with P21,Cyclin D1 and Cdk2.【Methods】 P73 gene were transfected into QBC939 cells.Cell 1ines of comparison was matched on the same genetic background and passage.The expression of wild-type P73,P21,Cyclin D1 and Cdk2 were detected by RT-PCR and Western blot.The cell growth and the cell cycle were examined by MTT and Flow Cytometry.【Results】 After transfection of QBC939 with transcription factor P73β,the expression of P21 mRNA was enhanced significantly.The expressions of cell cycle regulatory genes including Cyclin D1,cdk2 were decreased,and the inhibitory effects of P73β on ce11 growth were increased and the Cholangiocarcinoma cells grew slow.【Conclusion】 The wild-type P73β could increase the expression of P21 and decrease the expression of cyclin D1,Cdk2 and play an important role in S period.However,inhibit the proliferation of Cholangiocarcinoma QBC939 cells in vitro.