【作者】 田佳川；

【导师】 费晓方；

【作者基本信息】 吉林大学 ， 药物分析学， 2008， 硕士

【摘要】 本课题的研究目的是筛选具有抗肿瘤活性的药物,并在此基础上,进一步研究肿瘤细胞生长抑制的分子机制。通过体外生长抑制实验,结晶紫染色法测定细胞死亡率,我们发现从萝藦科(A sclep iadaceae)牛奶菜属(Marsdenia R.Br.)植物乌骨藤[M arsde ia tenacissin a (Roxb) .W ight etAm ]的藤茎中提取出的多种成分均可影响几种肿瘤细胞的生长。实验证明其含有的成分之一白桦酯酸(betulinic acid,BA)可以诱导HeLa细胞凋亡,且具有浓度和时间依赖性。通过以100μg/mLBA作用HeLa细胞48h后,对HeLa细胞总RNA进行提取后,进行RT-PCR反应,经琼脂糖凝胶电泳检测证明,BA可以诱导P21 mRNA和P53 mRNA表达的增高,细胞周期蛋白cyclinA mRNA变化不大,cyclinB1 mRNA表达降低。研究结果显示,白桦酯酸诱导肿瘤细胞凋亡可能与P53,P21基因改变有关,白桦酯酸可能进一步影响细胞周期的进程。更多还原

【Abstract】 Cell apoptosis is also named cell programmed death, which is one of the newest researches in biology. It is one of considerable vital phenomena in living nature. Apoptosis is closely associated with nemathelminth and high mammalia animal, embryo and adult, pathological and physiological conditions,life and die. It plays an important role in human health. As far as 100 years ago, Carl Vogt found this process. Kerr et al. have named it as“apoptosis”at 1972. Apoptosis is regulatoried by different genes, such as Ced gene family, bcl - 2 gene family, Fas/Apo gene family, P53 gene and ICE gene family etc.As one of Chinese traditional medicine, M arsdenia tenacissim a (Roxb) .Wight et Arn has been used for curing disease for thousands years in China. The factions and components were isolated from the fruits of M arsdenia tenacissim a (Roxb) .Wight et Arn. In our laboratory, those separate were screen for suppression activity of tumor cells. The result was shown that betulinic acid and betulin induced tumor cell died.This passage sieves medicine for curing tumor from constituents and chemical compounds are extracted from M arsdenia tenacissim a (Roxb) .Wight et Arn.This study on the apoptotic molecule mechanism in HeLa cell induced by betulinic acid. Do it with methods of crystal violet dying, RT-PCR were operated. Indicate the growth of tumor cells were significantly inhibited by betulin and betulinic acid in a time- and dose-dependent fashion. The furthers study indicated that betulinic acid induced HeLa cells apoptosis. The expression of P53 mRNA and P21 mRNA was increased and the expression of cyclinB1 mRNA was reduced in HeLa cells by Betulinic acid. But the expression of cyclinA mRNA was no change. The effect of Betulinic acid is mainly associated with apoptosis and cell cycle arrest.更多还原