| [Construction of the suppression subtractive cDNA libraries of human large cell lung cancer line L9981 before and after transfection with nm23-H1 gene.]. | |
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MedLine Citation:
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PMID: 20735954 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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BACKGROUND: It has been proven that nm23-H1 gene is an important metastaticsuppressed gene of lung cancer. In order to screen the differential expression genes related to nm23-H1 , we constructed the suppression subtractive cDNA libraries of human large cell lung cancer line L9981 transfected and untransfected with nm23-H1 gene by suppression subtractive hybridization (SSH) in this study, which lay a solid foundation for further screening and cloning metastatic-related genes of nm23-H1. METHODS: The forward and reverse suppression subtractive cDNA libraries were constructed in the human large cell lung cancer line L9981 before and after transfection with nm23-H1 gene (L9981 and L9981-nm23-H1) by SSH method. The positive clones were preliminarily screened by bluewhite colony, and precisely identified by PCR. RESULTS: The suppression subtractive cDNA libraries were successfully constructed in the human large cell lung cancer line L9981 transfected and untransfected with nm23-H1 gene (L9981-nm23-H1 and L9981). After the blue-white screening, about three hundred positive clones in the forward subtracted library and four hundred positive clones in the reverse subtracted library were obtained. Ramdom analysis of 96 clones in each library with colony PCR methods showed that 84 clones in the forward subtracted library and 83 clones in the reverse subtracted library contained (300-750) bp inserts. CONCLUSIONS: SSH is proved to be an efficient tool for differential expression gene cloning. The forward and reverse suppression subtractive cDNA libraries of human large cell lung cancer line L9981 transfected and untransfected with nm23-H1 gene (L9981-nm23-H1 and L9981) are successfully constructed by SSH and T/A cloning technology. The expression of nm23-H1 gene in the human large cell lung cancer cell lines may affect the differential expression of some metastatic-related genes. |
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Authors:
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Sujuan Ye; Zhihua Feng; Wen Zhu; Chunji Cai; Lu Li; Liya Sun; Haisu Wan; Li Ma; Qinghua Zhou |
Publication Detail:
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Type: English Abstract; Journal Article |
Journal Detail:
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Title: Zhongguo fei ai za zhi = Chinese journal of lung cancer Volume: 11 ISSN: 1999-6187 ISO Abbreviation: Zhongguo Fei Ai Za Zhi Publication Date: 2008 Aug |
Date Detail:
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Created Date: 2010-08-25 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101126433 Medline TA: Zhongguo Fei Ai Za Zhi Country: China |
Other Details:
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Languages: chi Pagination: 482-8 Citation Subset: - |
Affiliation:
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The State Key Laboratory of Biotherapy,West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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