为了探讨金雀异黄素(Genistein)对不同来源肿瘤生长的抑制作用以及主要抑癌作用机制,实验选择了体外培养的人神经母细胞瘤细胞SH-SY5Y,人红白血病细胞K562和人胃腺癌肝转移细胞NCI-N87作为研究对象.于细胞培养液中加入0.25-50 μg/mL的Genistein,通过MTT法发现Genistein能够抑制这三种肿瘤细胞的生长,半数杀伤浓度(IC50)分别为10.28 μg/mL、 19.83 μg/mL和40.59 μg/mL.荧光显微镜观察显示染色质固缩和断裂等凋亡特征.流式细胞术分析证明Genistein能够诱导这三种细胞产生凋亡,能够使K562和SH-SY5Y的细胞周期阻断在G2-M期.RT-PCR检测发现Genistein作用能够抑制这三种肿瘤细胞c-fos的表达.细胞粘弹性测定表明Genistein使K562细胞的弹性元素K1、K2和粘性系数μ明显增大,细胞刚性增大,变形能力降低.本实验为进一步的体内实验打下了基础,并为利用Genistein进行肿瘤的预防和治疗提供了有意义的实验依据.

The study is to transfer p16, an anti-oncogene for many tumors, into human Erythroleukemia cell line-K562 which is deleted with p16; and observe its expression and influence on rheologic behaviours of K562. First, through FuGene 6, p16 eukaryotic expressing vector pBABE-puro-p16 and empty vector pBABE-puro were introduced into K562, respectively. After 14 days, the positive cell colonies screened out by puromycin (1.5 μg/μL) were named as K562-p16 and K562-puro. The existence and expression of p16 in K562-p16 were confirmed by PCR, RT-PCR and Immunochemistry. The suppression of cell growth was observed in K562-p16. The G1-S arrest and apoptosis was discovered by FCAS. The electrophoretic mobility of K562-p16 increased significantly and K562 has better osmotic fragibility than K562-p16. All these show that exogenous p16 gene can suppress K562 growth in vitro, which is corroborated as G1-S arrest and apoptosis. Also p16 can induce the increase of surface charge and the drop of osmotic fragibility.

研究肿瘤坏死因子相关的凋亡诱导配体(TNF-related apoptosis inducing ligand,TRAIL)在四环素调控下的表达对人胃癌细胞NCI-N87的作用.将TRAIL的cDNA导入四环素调控系统RevTet-On,用逆转录病毒感染的方法在胃癌细胞系NCI-N87中建立四环素调控TRAIL基因表达系统及对照系统.用四环素衍生物强力霉素诱导基因表达,检测其对胃癌细胞的杀伤作用.获得了在人胃癌细胞系NCI-N87中建立的四环素调控TRAIL基因表达系统,在强力霉素诱导后观察到了细胞的死亡.证实四环素调控TRAIL表达系统对胃癌细胞具有杀伤作用.

1 介绍 1.1 蛋白质组的概念 二十世纪七十年代以来,对于生命体的研究集中在基因组学和基因工程.1988年发起的人类基因组规划(the human genome project,HGP),大大推动了分子生物学的发展,在不久的将来,所有分子水平上的遗传信息便会被破译.