Studvofbioloeical function of CD10/NEP24.11 protein on the B-cells

Research Project

Project/Area Number	17590310
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Single-year Grants
Section	一般
Research Field	Human pathology
Research Institution	Fukushima Medical University
Principal Investigator	ABE Masafumi Fukushima Medical University, School of Medicine, professor (00045783)
Co-Investigator(Kenkyū-buntansha)	TASAKE Kazuhiro Fukushima Medical University, School of Medicine, associated professor (70244382) SUZUKI Osamu Fukushima Medical University, School of Medicine, assistant professor (00325953) 野沢佳弘福島県立医科大学, 医学部, 講師 (40208343)
Project Period (FY)	2005 – 2007
Project Status	Completed (Fiscal Year 2007)
Budget Amount *help	¥3,800,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥300,000) Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000) Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywords	CD10 / NEP24.11 / endothelin A receptor / NEP24.11 tranfectant / cell proliferation / human lymphoma cell line / NEP24.11酵素活性 / ヒトB細胞リンパ腫細胞株 / RT-PCR / ウエスタンブロット法 / Endothelin A receptor / フローサイトメトリー
Research Abstract	CD 10/neutral endopeptidase (NEP)24.11 expression was detected in 9 B-cell lymphoma cell lines (5 Burkitt lymphoma cell lines, 2 follicular lymphoma cell lines, 1 immunoblastic lymphoma cell line and 1 plasma cell myeloma cell line) out of 17 human lymphoma cell lines by flow cytometry, Western blot and RT PCR methods, and the enzymatic activity was confirmed by a fluorometric assay. The presence of CD10/NEP24.11 protein on the surface of B-cell lymphoma cells suggests that the enzymatic activity play an important role in the biological characteristics of lymphoma cells. Overexpression of CD10/NEP24.11 protein was suggested to require mRNA expression of exon 1, exon 2a and exon 2b and exon 3 and exon 2a might regulate overexpression of CD10/NEP24.11 protein. Expression of a variety of receptors for CD10/NEP24.11 substrates was analyzed using Western blot and RT姫CR methods. An only follicular lymphoma cell line FL-18 expressed both mRNA and protein of endothelin receptor A (ETAR). However ; endothelin-1 did not affect cell proliferation of FL-18 lymphoma cells. CD10/NEP24.11 stable transfectants of the CD10/NEP24.11 negative human diffuse large B-cell lymphoma cell line HBL-1 were generated, and the mRNA and protein expression of CD10/NEP24.11 was confirmed using immunocytochemistry, Western blot and RT-PCR methods. The enzymatic activity was demonstrated by a fluorometric assay. There was a trend towards increased cell proliferation in CD10/NEP24.11 transfectants of HBL-1 cells compared to CD10/NEP24.11 negative HBL-1 cells. We will perform in vitro invasion assay, in vitro migration assay, tumor progression in vivo and identification of genes that co-express with CD10/NEP24.11 using CD10/NEP24.11 transfectants of HBL-1 cells to elucidate biological functions of CD10/NEP24.11.