【作者】 金焰； 李明； 吴杰； 吕娜； 于旸； 白静； 陈峰； 傅松滨；

【Author】 Laboratory of Medical Genetics,Harbin Medical University,Harbin,Heilongjiang,150081,P.R.China.)

【机构】 哈尔滨医科大学基础医学学院医学遗传学研究室；

【摘要】 目的 :p53基因是重要的抑癌基因之一。它抑制肿瘤细胞增殖、参与细胞凋亡调控,但具体机制尚不十分清楚。本研究采用凋亡相关基因芯片检测p53基因介导肺癌细胞发生凋亡的通路,拟阐明与之相关的调控机制。方法 :采用Superarry公司的凋亡检测芯片分别对转染p53前后的四种肺癌细胞系中凋亡相关基因的表达进行检测。通过体外细胞培养,普通光镜、电子显微镜、流式细胞计数法、MTT比色法,siRNA和western blot等体外实验,分别对芯片中阳性基因功能鉴定分析。结果 :感染Ad-p53后,BNIP3基因在高转移细胞系Anip973和95D中的表达量均显著提高,而在低转移细胞系AGZY83-a和95C中表达改变不明显。分别构建BNIP3基因的真核正义表达载体和siRNA,在高转移细胞系Anip973中对该基因进行反义沉默,在低转移细胞系AGZY83-a中过表达该基因,证实了两种条件下均对细胞形态和生长趋势产生一定的影响。而且,分别对上述两种细胞过表达p53后,发现过表达BNIP3的AGZY83-a细胞的凋亡作用得以增强,而对沉默了BNIP3的Anip973细胞的凋亡作用有所下降。结论 :BNIP3参与了p53介导的肺癌细胞凋亡的过程,这可能也是p53介导高转移肺癌细胞系快速凋亡的机制之一。更多还原

【Abstract】 Objective It has been shown that p53 plays key roles in induction of cell apoptosis.However,the molecular mechanism of the implicated pathway is still unclear.In this study,we identified apoptosis related genes that were downstream of p53.Methods The p53 was overexpressed or knock down in lung cancer cells.Then superarray chip were used to detect the expression of apoptosis related genes.Molecular biology methods were employed to check the function of these genes.Results Here,we show that overexpression of exogenous wild-type p53 induced apoptosis in lung cancer cells.High metastasis potential cells had a faster rate of apoptosis than low metastasis potential cells.The expression of proapoptotic gene BNIP3 was increased significantly both in Anip973 and 95D cell lines which have high metastasis ability,but not in AGZY83-a or little increased in 95C cell lines which possess low metastasis ability.Overexpression of BNIP3 increases apoptotic rate induced by p53 in AGZY83 -a cells.Blocking the expression of BNIP3 by siRNA in Anip973 cells decreased apoptotic rate mediated by p53.Conclusions Taken together,these data suggested that high level expression of BNIP3 mediated rapid apoptosis which was triggered by p53 in lung cancer cells.更多还原