力达霉素对人肝癌bel-7402细胞凋亡基因表达和细胞骨架的影响  

Effects of Lidamycin on Apoptotic Gene Expressions and Cytoskeleton in Human Hepatoma bel-7402 Cells

作　　者：何琪杨[1] 姜兵[1] 李电东[1] 甄永苏[1] 

机构地区：[1]中国医学科学院中国协和医科大学医药生物技术研究所,北京100050

出　　处：《癌症》2002年第4期351-355,共5页Chinese Journal of Cancer

基　　金：国家自然科学基金项目(编号30070878)

摘　　要：背景与目的:力达霉素是我所分离的烯二炔类抗肿瘤抗生素,因其独特的化学结构和对体内外肿瘤的强烈抑制作用,而倍受关注。除了断裂DNA外,力达霉素对细胞凋亡基因表达和细胞骨架的影响,可能与其高活性有关。本文通过观察力达霉素对人肝癌bel-7402细胞多指标的影响,进一步阐明其分子机制。方法:用Northernblot印迹法和dot印迹法检测癌基因表达,间接免疫荧光法检测微丝和微管,线粒体特异染料MitosensorTM检测细胞凋亡。结果:低浓度的力达霉素(0.1nmol/L)处理人肝癌bel-7402细胞8h,明显促进c-myc、c-fos基因表达,而抑制N-ras表达。10nmol/L力达霉素处理细胞8h后,细胞伸展,微丝排列整齐,向非恶性细胞转变,但对微管没有影响。用线粒体特异凋亡染料MitosensorTM检测发现,力达霉素处理8h后的细胞未出现凋亡,说明力达霉素引起肿瘤细胞凋亡首先需要诱导c-myc基因表达。结论:低浓度的力达霉素明显影响人肝癌bel-7402细胞的有关凋亡基因表达和细胞骨架的分布,这些结果有助于解释力达霉素高活性地作用肿瘤细胞的分子机制。Background &Objective:Lidamycin, one of enediyne antitumor antibiotics, was isolated by our institute. It is the focus of intense due to its unique chemical structure and potent cytotoxicities to tumor cells in vivo and in vitro. In addition to cleavage of DNA, effect of lidamycin on apoptotic gene expressions and cytoskeleton may involve in its high activities. To further elucidate its mechanism, we have observed the effect of lidamycin on the above mentioned factors in human hepatoma bel 7402 cells. Methods:The gene expressions were determined by Northern blot and dot blotanalysis. The changes of microfilament and microtubule were detected by indirect immunofluorescent method and the apoptotic cells were detected with mitochondria specific apoptotic dye MitosensorTM. Results:The obvious increment of c myc and c fos gene expressions and the inhibition of N ras gene expression in bel 7402 cells were observed following lidamycin treatment (0.1-10 nmol/L) for 8 h. The arrangement of microfilament in the cells became regular and was similar to nonmalignant normal cells, but there was no effect on microtubule when the bel 7402 cells were treated with lidamycin 10 nmol/L for 8 h. No apoptotic cells were detected with MitosensorTM in the lidamycin treated cells for 8 h. Conclusion:Lidamycin at lower concentrations can markedly affect the expression of the apoptosis related genes and the distribution of the cytoskeleton in the hepatoma bel 7402 cells. The results are helpful to elucidate the molecular mechanism of potent cytotoxicities of lidamycin to tumor cells.

关 键 词：力达霉素 基因表达 细胞骨架 凋亡 人肝癌细胞 

分 类 号：R735.7[医药卫生—肿瘤]