复方木鸡冲剂诱导人白血病细胞HL-60凋亡机制研究

目的:探讨复方木鸡冲剂诱导人白血病细胞HL-60细胞凋亡的作用和机制,为相关中药开发提供实验资料.方法:用MTT法检测复方木鸡冲剂对HL-60细胞增殖活性的影响,光镜下观察细胞形态的变化;流式细胞仪(Annexin V/PI双染法)检测细胞凋亡,并分析细胞周期;免疫细胞化学法检测Bcl-2、caspase-3、p21WAF1的表达.结果:MTT法显示复方木鸡冲剂能抑制HL-60细胞的生长,细胞呈凋亡形态学变化.流式细胞仪检测结果为细胞凋亡率明显增高,出现GO/G1期阻滞.Bcl-2表达降低,caspase-3表达增高,p21WAF1表达强阳性.结论:复方木鸡冲剂明显抑制HL-60细胞的生长,其抗肿瘤的机制与诱导肿瘤细胞凋亡、促进细胞分化有关.     

核转录因子相关因子2的活化在肿瘤代谢中的作用

核转录因子(NF-E2)相关因子2（nuclear factor erythroid 2 related factor 2, Nrf2）是细胞应对外界应激的主要调控因子,通过调控多种靶基因的表达,在生理条件下减轻氧化应激,维持细胞稳态。其上游受多种因素调控,包括氧化与亲电应激、外界营养状态、细胞内代谢中间产物和能量状态等。在肿瘤细胞中,异常活跃的Nrf2使其抗氧化能力增强,并且通过介导代谢重编程（metabolic reprogramming）,促进肿瘤细胞增殖和生长。Keap1 (Kelch-like ECH-associated protein 1)是氧化和亲电应激感受器,通过募集泛素降解系统,对Nrf2的活性起主要调控作用。本文介绍Keap1依赖与非依赖条件下Nrf2的活化途径,着重介绍在肿瘤中Nrf2的异常活化,以及如何调控代谢重编程进而调节肿瘤细胞的合成代谢,最终促进肿瘤的进展。     

Adenovirus-mediated transfer of RA538 gene and its antitumor effect

The RA538 cDNA was transferred into human ovarian cancer cell line SK-OV-3 and human melanoma cell line WM-983A by its recombinant adenoviral vector constructed through homologous recombination. It was demonstrated that the recombinant adenovirus could transfer RA538 gene with high efficiency, and could obviously inhibit tumor growth, with the inhibiting rates of 85% and 73% respectively, at the same time greatly repress the colony forming ability of the cells. The therapeutic experiments on transplanted subcutaneous tumor model in nude mice demonstrated that RA538 could significantly inhibit tumor growth. Flow cytometry and DNA fragmentation analysis indicated that RA538 could induce the cell cycle G1 arrest/apoptosis of the tumor cells. The expression of cmyc gene was found pronouncedly reduced by Western blot analysis. These results suggest that the RA538 recombinant adenovirus could be a promising drug in cancer gene therapy.     

腺病毒介导RA538基因转移及其肿瘤抑制作用

采用同源重组方法构建RA5 3 8cDNA重组体腺病毒 ,通过转导人卵巢癌细胞系SK OV 3和人黑色素瘤细胞系WM 983A ,证实腺病毒可高效转导RA5 3 8基因 ,能显著抑制肿瘤细胞的生长 ,其抑制率分别达 85 %和 73 % ;并显著降低两细胞系的集落形成能力 .对两细胞系的裸鼠皮下移植瘤进行治疗实验结果表明 ,RA5 3 8能明显抑制肿瘤的生长 .流式细胞计数和DNA片段化分析证实 ,RA5 3 8可引起肿瘤细胞G1期阻滞和 /或凋亡 .对c myc蛋白的Westernblot分析发现 ,RA5 3 8有明显下调c myc基因表达的作用 .根据这些结果 ,我们认为RA5 3 8重组体腺病毒有可能成为有应用价值的肿瘤基因治疗药物 .