(A) novel alternative splicing form of Met reduces the tumorigenic potential of Met protooncogene. = 새로운 간세포 성장인자 수용체 변이형의 암화 억제 능력

목차 (Table of Contents)

• TABLE OF CONTENTS
• ABSTRACT----------------------------------------------------------------------------------------------ⅰ
• TABLE OF CONTENTS-----------------------------------------------------------------------------ⅲ
• LIST OF FIGURES------------------------------------------------------------------------------------ⅴ
• LIST OF TABLES-------------------------------------------------------------------------------------ⅶ
• Ⅰ. INTRODUCTION----------------------------------------------------------------------------------1
• A. Structure of Met receptor tyrosine kinase------------------------------------------------------------1
• B. Aberrant activation of HGF-Met signaling in cancers---------------------------------------------3
• C. Therapeutic strategies targeting Met in human cancers-------------------------------------------9
• D. HGF-Met signaling in muscle differentiation and maintenance of rhabdomyosarcoma--11
• E. Different splicing variants of Met---------------------------------------------------------------------13
• F. D13Met, a novel alternative splicing variant of Met -----------------------------------------15
• Ⅱ. MATERIALS AND METHODS-----------------------------------------------------------------17
• A. Cell culture and transfections --------------------------------------------------------------------17
• B. Reagents and antibodies--------------------------------------------------------------------------------18
• C. RT-PCR----------------------------------------------------------------------------------------------------19
• D. Immunoprecipitation and Western blotting--------------------------------------------------------19
• E. Cell proliferation and thymidine incorporation assay--------------------------------------------20
• F. Soft-agar colony formation assay----------------------------------------------------------------------21
• G. In vivo tumorigenesis assay----------------------------------------------------------------------------21
• H. Colony generation assay--------------------------------------------------------------------------------22
• I. Migration assay--------------------------------------------------------------------------------------------22
• J. Differentiation induction of primary cell or rhabdomyosarcoma cell lines------------------23
• K. Lentiviral vector production------------------------------------------------------------------------------23
• L. Statistical analysis------------------------------------------------------------------------------------------24
• Ⅲ. RESULTS-------------------------------------------------------------------------------------------25
• Part. 1. Inhibition of HGF-Met signaling by 13Met -------------------------------------------25
• A. 13Met inhibits Met activation induced by HGF -----------------------------------------------25
• B. 13Met inhibits HGF-Met signaling-dependent cell proliferation and colony
• generation-----------------------------------------------------------------------------------------------------28
• C. 13Met inhibits tumor growth in mice-------------------------------------------------------------32
• D. Status of HGF-Met signaling is variable in rhabdomyosarcoma cells-----------------------34
• E. Met silencing induces inhibition of ERK phosphorylation in RD cells-----------------------36
• F. D13Met inhibits HGF-Met signaling in RH18 and RD cells------------------------------------38
• G. Δ13Met reduces colony formation in RD cells----------------------------------------------------40
• H. Δ13Met inhibits migration of RD cells-------------------------------------------------------------42
• Part. 2. Induction of differentiation by D13Met ----------------------------------------------44
• I. D13Met is induced during muscle differentiation-------------------------------------------------44
• J. Abolishment of D13Met induction with specific siRNA results in delayed
• differentiation of primary human skeletal muscle cells-------------------------------------------- 48
• K. Overexpression of D13Met enhances differentiation in primary human skeletal muscle
• cells---------------------------------------------------------------------------------------------------------51
• L. Δ13Met is unable to induce differentiation of RD cells------------------------------------------54
• M. Ectopic expression of D13Met hardly increases MHC expression in several Rhabdo-
• myosarcoma cell lines under differentiation condition------------------------------------56
• Ⅳ. DISCUSSION---------------------------------------------------------------------------------------58
• Ⅴ. CONCLUSION-------------------------------------------------------------------------------------63
• BIBLIOGRAPHY--------------------------------------------------------------------------------------64
• 국문요약-------------------------------------------------------------------------------------------------74