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、、和在NIH3T3细胞致瘤转化中的作用。

Roles of , , and in tumourigenic transformation of NIH3T3 cell.

作者信息

Chang Cuifang, Xi Lingling, Zhang Jihong, Zhao Weiming, Liu Zhiyou, Guo Jianlin, Xu Cunshuan

机构信息

State Key Laboratory Cultivation Base for Cell Differentiation Regulation, Henan Normal University, Xinxiang 453007, People's Republic of China.

出版信息

J Genet. 2018 Dec;97(5):1155-1168.

Abstract

To analyse the mechanism of tumourigenic transformation of NIH3T3 cells at the transcriptional level, we used cancerogen 3-methylcholanthrene (3-MCA) and cancerogenic substance phorbol-12-myristate-13-acetate (PMA) to transform NIH3T3 cells and the assessment of transformation was performed using Giemsa staining and methylcellulose colony formation assay. Changes in gene expression profile were detected by Mouse Genome 430 2.0 microarray; and quantitative real-time polymerase chain reaction and Western blotting were used to verify the expression changes of mRNAs and proteins, respectively. With the aid of bioinformatics method, five signalling pathways were identified to participate in different stages of NIH3T3 cell transformation. Further, our study suggested that oncogenes , , and the tumour suppressor gene may play important roles in these pathways.

摘要

为了在转录水平分析NIH3T3细胞致瘤转化的机制,我们使用致癌物3-甲基胆蒽(3-MCA)和致癌物质佛波酯-12-肉豆蔻酸酯-13-乙酸酯(PMA)来转化NIH3T3细胞,并使用吉姆萨染色和甲基纤维素集落形成试验进行转化评估。通过Mouse Genome 430 2.0芯片检测基因表达谱的变化;分别使用定量实时聚合酶链反应和蛋白质免疫印迹法来验证mRNA和蛋白质的表达变化。借助生物信息学方法,鉴定出五条信号通路参与NIH3T3细胞转化的不同阶段。此外,我们的研究表明,癌基因 、 、 和肿瘤抑制基因 可能在这些通路中发挥重要作用。

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