摘要
背景与目的核因子κB(NFκB)参与多种肿瘤的发生发展,抑制其活性可能抑制肿瘤细胞生长。本研究旨在观察抑制NFκB对人肺癌细胞株H460生长的影响,探讨其抑制或杀伤肿瘤的分子机制。方法体外实验:用表达抑制性κB(IκBα)或LacZ的腺病毒(AdIκBα或AdLacZ)转染H460细胞,测定其生长和凋亡情况,并检测细胞培养上清液中caspase3和TNFα表达。体内实验:用预先转染AdIκBα/AdLacZ的H460细胞建立肿瘤模型,以观察基因转染对成瘤性的影响;对未转染的H460肿瘤小鼠,于瘤内注射AdIκBα/AdLacZ/PBS,观察小鼠肿瘤的生长。留取小鼠肿瘤标本进行免疫组化染色,检测p65、p53和VEGF表达。结果①转染AdIκBα可抑制肿瘤细胞生长,促进凋亡发生;②成瘤性抑制实验显示接种30MOIAdIκBα转染的H460细胞的小鼠中77.8%在6周内无肿瘤形成,而30MOIAdLacZ转染组仅12.5%(P=0.012);③对已建立的H460肿瘤,瘤内注射30MOIAdIκBα可明显延缓肿瘤生长(P<0.05);④免疫组化染色提示各实验组的p53表达无显著性差异,而VEGF表达在30MOIAdIκBα转染组明显减少,转染AdIκBα抑制了H460细胞的p65活性。结论AdIκBα转染阻滞人肺癌细胞H460的NFκB活性,抑制肿瘤细胞的成瘤性,并延缓肿瘤生长。
Background and objective Nuclear factor κB (NFκB) is involved in oncogenesis and development. Inhibiting its activity may affect cancer cell growth. The aim of this study is to investigate the effects of inhibitor κB transfection on proliferation and tumorigenesis of human lung cancer cell line H460. Methods H460 cells were transfected with adenovirus expressing inhibitor κBα or LacZ (AdIκBα/AdLacZ) and cell viability was detected. Apoptosis was detected by Annexin-V/PI stain and caspase 3 activity measurement. In (vivo) tumorigenesis test was performed in nude mice inoculated with transfected H460 cells. The therapeutic effect of AdIκBα was evaluated in the inoculated tumors. p65, p53 and VEGF expressions were detected in implanted tumor tissues by immunohistochemical stain. Results AdIκBα transfection significantly inhibited the proliferation of H460 cells. Apoptosis and expression level of caspase 3 in 30 MOI AdIκBα transfection group were remarkably higher than those in control groups (P<0.05). In vivo study showed tumor incidence was significantly lower in 30 MOI AdIκBα transfection group than that in virus control group (P=0.012). For established H460 tumors, intratumoral injection of AdIκBα could significantly inhibit the tumor growth. Immunohistochemical stain showed VEGF expression was decreased in 30 MOI AdIκBα transfection group and p65 activity was blocked in H460 cells by AdIκBα transfection. Conclusion AdIκBα transfection can inhibit the growth of H460 cells, as well as tumorigenesis. It might be due to both blockade of NFκB activity and inhibition of VEGF expression, which would be independent of p53 expression.
出处
《中国肺癌杂志》
CAS
2005年第3期175-180,共6页
Chinese Journal of Lung Cancer
