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目的:探讨shRNA介导的神经生长因子β(NGF-β)下调对人胆管癌细胞系QBC939体外增殖与凋亡的影响。方法:构建稳定转染NGF-βshRNA的QBC939细胞系,将细胞分为干扰组(转染NGF-βshRNA组)和对照组(转染空质粒组)。以Western blot方法检测细胞转染效果;细胞增殖实验(CCK-8实验)检测细胞增殖能力,单克隆形成实验检测细胞克隆形成能力;流式细胞术检测细胞周期及细胞凋亡。结果:干扰组细胞较之对照组的细胞增殖、克隆形成能力明显降低,S期细胞明显减少(P<0.05),凋亡率明显增加(P<0.05)。结论:抑制NGF-β能降低QBC939细胞系的增殖和克隆形成能力,该作用与其阻滞细胞进入S期,促进细胞凋亡有关。
AIM: To investigate the effect of shRNA-mediated down-regulation of nerve growth factor β (NGF-β) on proliferation and apoptosis of human cholangiocarcinoma cell line QBC939 in vitro. Methods: QBC939 cells stably transfected with NGF-βshRNA were constructed and divided into interference group (transfected with NGF-βshRNA group) and control group (transfected with empty plasmid group). Cell transfection was detected by Western blot. Cell proliferation assay (CCK-8 assay) was used to detect cell proliferation ability. Monoclonal formation assay was used to detect cell clonality. Cell cycle and apoptosis were detected by flow cytometry. Results: Compared with the control group, the clonal formation ability of the cells in the interference group was significantly decreased, the cells in the S phase were significantly decreased (P <0.05), and the apoptosis rate was significantly increased (P <0.05). Conclusion: Inhibition of NGF-β can reduce the proliferation and colony formation ability of QBC939 cell line, which may be related to arresting cells entering S phase and promoting cell apoptosis.