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标题：Forward genetic screens in mice uncover mediators and suppressors of metastatic reactivation
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作者：Hua Gao ; Goutam Chakraborty ; Ai Ping Lee-Lim 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2014
卷号：111
期号：46
页码：16532-16537
DOI：10.1073/pnas.1403234111
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：SignificanceInsights into the mechanisms that enable disseminated cancer cells to survive during dormancy and then outgrow into life-threatening lesions may lead to the identification of novel therapeutic targets for the prevention or treatment of metastatic disease. We have developed flexible and high-throughput functional genetic screens, which enable the identification of single genetic entities that mediate metastatic reactivation of breast cancer in mice. These screens promise to facilitate the identification of the core signaling pathways that govern metastatic dormancy and reactivation. We have developed a screening platform for the isolation of genetic entities involved in metastatic reactivation. Retroviral libraries of cDNAs from fully metastatic breast-cancer cells or pooled microRNAs were transduced into breast-cancer cells that become dormant upon infiltrating the lung. Upon inoculation in the tail vein of mice, the cells that had acquired the ability to undergo reactivation generated metastatic lesions. Integrated retroviral vectors were recovered from these lesions, sequenced, and subjected to a second round of validation. By using this strategy, we isolated canonical genes and microRNAs that mediate metastatic reactivation in the lung. To identify genes that oppose reactivation, we screened an expression library encoding shRNAs, and we identified target genes that encode potential enforcers of dormancy. Our screening strategy enables the identification and rapid biological validation of single genetic entities that are necessary to maintain dormancy or to induce reactivation. This technology should facilitate the elucidation of the molecular underpinnings of these processes.
关键词：forward genetic screens ; metastatic reactivation ; cDNA library screen ; shRNA library screen ; microRNA library screen