国家自然科学基金资助项目(30370338),天津市科技发展计划科技发展计划科技攻关项目(043182911).
This work was supported by grants from The National Natural Science Foundation of China (30370338) and Key Project of Development for Science and Technology in Tianjin (043182911).
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 21 329 个基因的芯片进行杂交并扫描,利用 GenePix Pro 4.0 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 21 329 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, apoptosis, 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.
叶丽虹,尤嘉琮,乔 玲,蔡 娜,王长晔,刘 毅,吴晶辉,吴莲英,王洪辉,张晓东.应用基因表达谱芯片从不同转移能力的 乳腺癌细胞中筛选转移相关基因[J].生物化学与生物物理进展,2005,32(5):421-428
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