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学科主题: 基础医学
题名:
Monoclonal antibodies against human tumor metastasis suppressor gene-1 (TMSG-1): Preparation, characterization, and application
作者: Pei, F; Ning, JY; You, JF; Yang, JP; Wang, YP; Han, ZH; Wang, JL; Cui, XL; Yang, SM; Zheng, J
刊名: HYBRIDOMA AND HYBRIDOMICS
发表日期: 2004-10-01
卷: 23, 期:5, 页:318-325
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemical Research Methods ; Biotechnology & Applied Microbiology ; Immunology
研究领域[WOS]: Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Immunology
关键词[WOS]: LONGEVITY-ASSURANCE GENE ; LAG1 ; YEAST ; CLONING ; CANCER ; LAC1P
英文摘要:

To better understand the molecular mechanism of metastasis, the monoclonal antibody against tumor metastasis suppressor gene - 1 (TMSG-1, one novel gene) was prepared, characterized, and applied in estimating the metastatic potential of human tumors. A dominant epitope, TMSG-1(15) - derived from TMSG-1 - was automatically synthesized based on the Fmoc method, and the hapten was conjugated to Imject Maleimide activated mcKLH as a carrier protein. The antigen solution was used to immunize BALB/C mice. Hybridomas were generated and screened by ELISA for specific monoclonal antibodies, and their characterization was performed by Western blotting and immunohistochemical staining. The hybridoma cell line secreting anti - TMSG-1 antibody, designated C8, was established after primary ELISA screening and consequent rapid limited dilution. C8 was IgM in isotyping. The competitive inhibition assay showed that the antibody was TMSG-1 specific. Furthermore, in Western blotting, a protein band of about 45 kD was detected with non-metastatic variant PC-3M-2B4 and PG-LH7 cells, but not with the isogenetic metastatic variant PC-3M-1E8 and PG-BE1 cells. Immunohistochemistry method showed that positive staining presented in the cytomembrane and cytoplasm of 2B4 and LH7 cells, while 1E8 and BE1 cells were non-reactive. The sections from paraffin-embedded blocks of human cancer ( 52 cases of breast carcinoma and 41 cases of colon cancer) were stained, showing strongly positive in non-metastatic tumor and weakly positive or negative in metastatic tumor; the strongly positive rate of TMSG-1 expression in human cancers were, respectively, 36%, 7.4%, 52.4%, and 35% in non-metastatic and metastatic breast cancer and non-metastatic and metastatic colon cancer ( p = 0.02). The monoclonal antibody developed against synthetic peptide was TMSG-1 specific, and it has promise in Western blot and immunohistochemistry for detecting the TMSG-1 expression of cancer cells and tissues. It may provide an important tool in the study of TMSG-1 expression and function in both experimental and clinical studies. Furthermore, our tests confirmed that TMSG-1 protein has excellent inverse correlation to tumor metastasis potential, with the molecular weight of 45 kD ( supporting the encoded protein containing 380 amino acids) and localization in cytomembrane and cytoplasm of tumor cell.

语种: 英语
WOS记录号: WOS:000224843200008
Citation statistics:
内容类型: 期刊论文
版本: 出版稿
URI标识: http://ir.bjmu.edu.cn/handle/400002259/58336
Appears in Collections:基础医学院_病理学系_期刊论文

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作者单位: Peking Univ, Hlth Sci Ctr, Dept Pathol, Beijing 100083, Peoples R China

Recommended Citation:
Pei, F,Ning, JY,You, JF,et al. Monoclonal antibodies against human tumor metastasis suppressor gene-1 (TMSG-1): Preparation, characterization, and application[J]. HYBRIDOMA AND HYBRIDOMICS,2004,23(5):318-325.
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