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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.
The human breast cancer cell line MCF-7 and its metastatic subclone LM-MCF-7 provide a cellular model for studying the molecular mechanisms of tumor metastasis.Comparing the difference of gene expression profiles of two cell lines with different metastatic potential using gene chip technology, The cDNA of LM-MCF-7 and MCF-7 were respectively labeled with Cy5-dCTP and Cy3-dCTP, and then hybridized with the 21329 gene chip and scanned. GenePixPro4 .0 image analysis software processing data to determine whether the gene expression difference between the two cells.Transcribed fluorescent markers repeated two experiments, a total of 67 differentially expressed genes were screened, of which 41 in LM-MCF-7 cells Up-regulated, and 26 down-regulated in LM-MCF-7 cells.At the same time, seven genes with significant differences were verified by real-time quantitative RT-PCR.The bioinformatics analysis indicated that the above- Internal signal transduction, transcriptional regulation, stress response, metabolism, development, cell motility, apoptosis and cell adhesion.According to the literature, 35 of these differentially expressed genes are associated with tumor , Of which 9 were related to breast cancer metastasis and 6 were involved in tumor invasion and metastasis.According to the results of gene chip test, the relationship between LM-MCF-7 cells and MCF-7 cells and apoptosis were functionally studied The results showed that the anti-apoptotic ability of LM-MCF-7 cells with high metastatic potential was stronger than that of MCF-7 cells.Therefore, the role and molecular mechanisms of the above-mentioned genes related to tumor metastasis need to be further studied.