腺病毒介导的Bcl-XL shRNA对结肠癌细胞的体外抗肿瘤性

Anti-tumor effect of adenovirus-mediated BcI-XL shRNA in vitro

目的研究腺病毒介导的Bcl—XL shRNA对结肠癌细胞株的体外抗肿瘤性。方法将已有的Bcl-XL腺病毒进行扩增纯化．并通过反转录反应分析该腺病毒在mRNA水平对Bcl-XL的表达调控。通过MTT检测及克隆形成实验，了解该腺病毒对结直肠癌细胞株（Lovo细胞株）的体外杀伤作用。结果Ad／Bcl．XLshRNA作用于Lovo细胞株导致Bcl—XL mRNA表达水平下降。与对照腺病毒Ad／GFP相比，Ad／Bcl-XL shRNA显著抑制Lovo细胞的增长（P〈0．05），且该抑制效应呈剂量和时间依赖性；显著抑制Lovo细胞的克隆形成（P〈0．05），且该抑制效应呈剂量依赖性。而Ad／Bcl—XL shRNA对正常人成纤维细胞（NHFB）无明显杀伤作用。结论腺病毒介导的Bcl-XL shRNA对Lovo细胞株具有杀伤作用，可能存在潜在的结肠癌治疗价值。

Objective To investigate the anti-tumor effect of adenovirus-mediated Bcl-XL shRNA on colon cancer cells in vitro. Methods A recombinant Bcl-xl adenovirus was constructed, applified, and purified. The effect on mRNA expression of Bcl-XL was assessed by RT-PCR, and the effect on apoptosis-induction of colon cancer （Lovo cell line） in vitro was assessed by MTF assay and cell clonogenic assay. Results RT-PCR showed that Ad/Bcl-XL shRNA significantly down-regulated the mRNA expression of Bcl-XL in Lovo cells. Ad/Bcl-XL shRNA suppressed the proliferation of Lovo cells in a dose-dependent as well as a time-dependent manner compared with Ad/GFP（P〈0.05）. Treatment with Ad/Bcl-XL shRNA dramatically suppressed the colony formation of Lovo cells in a dose- dependent manner （P〈0.05）. Ad/Bel-XL shRNA showed no effect on normal human fibroblast. Conclusion Ad/Bcl-XL shRNA exhibits cytotoxic effect on Lovo cells and may have the potential value in the treatment of colon cancer.