摘要
人乳腺癌细胞系MCF-7及其转移亚克隆LM-MCF-7为肿瘤转移分子机制的研究提供了细胞模型.应用基因芯片技术比较两种具有不同转移能力细胞系基因表达谱的差异,寻找乳腺癌转移相关基因.提取两种细胞总RNA,分别用Cy5-dCTP、Cy3-dCTP标记LM-MCF-7和MCF-7的cDNA,并与含有21329个基因的芯片进行杂交并扫描,利用GenePixPro4.0图像分析软件处理数据判断基因是否在两个细胞中存在表达差异.经互换荧光标记物重复两次实验,共筛选出差异表达基因67个,其中41个在LM-MCF-7细胞中表达上调,26个在LM-MCF-7细胞中表达下调.应用实时定量RT-PCR对7个表达差异明显的基因进行了验证.生物信息学分析结果提示,上述发现的差异基因编码产物与细胞内信号转导、转录调节、应激反应、新陈代谢、发育、细胞运动、细胞凋亡和细胞粘连等功能有关.据文献报道,这些差异表达的基因中有35个与肿瘤有关,其中9个与乳腺癌转移有关,6个可能参与肿瘤浸润和转移过程.根据基因芯片检测的结果,从功能上对LM-MCF-7细胞和MCF-7细胞与细胞凋亡的关系进行了研究,发现具有高转移倾向的LM-MCF-7细胞与MCF-7细胞相比,抗凋亡能力较强.上述与肿瘤转移相关基因在肿瘤转移中的作用及其分子机理有待深入研究.
Human breast cancer cell line MCF-7 and its sub-clone LM-MCF-7 with high metastatic potential supply a cell model for studying molecular mechanisms of tumor metastasis. The two cell lines were used to screen tumor metastasis-associated genes by cDNA microarray, displaying differential gene expression profile. Total RNA of the two cells was extracted and the cDNAs were labeled with Cy5-dCTP and Cy3-dCTP, respectively. Then the cDNAs were hybridized on the gene chips containing 21329 kinds of human genes. The signals were examined by GenePix Pro 4.0 software. The two labeling nucleic fluorescent dyes were exchanged each other so that the experiments were repeated twice. The results shown that 67 kinds of genes were markedly different between the two cells, in which 41 were up-regulated and 26 down-regulated in LM-MCF-7 cells. Some of results were confirmed by real-time PCR. The above genes. are associated with functions, such as cell signal transduction, transcript regulation, transportation, response to stress, metabolism, development, movement, cell cycle, differentiation, apopto is, conglutination, and so on. According to the reports 35 of 67 genes were involved in tumor, in which 9 were related to metastasis of breast tumor, and 6 may be involved in tumor invasion and metastasis. Based on the results, apoptosis was investigated in function in LM-MCF-7 and MCF-7 cells. It was found that the ability of antiapoptosis in LM-MCF-7 cells was higher than that in MCF-7 cells. The candidate genes associated with tumor metastasis will be further demonstrated in function.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2005年第5期421-428,共8页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金资助项目(30370338)
天津市科技发展计划科技攻关项目(043182911)~~
关键词
基因芯片
基因表达谱
乳腺癌
肿瘤转移
gene chips
gene expression profile
breast cancer
tumor metastasis
