A study on the role of discoidin domain receptors in cancer cell migration and invasion
- 주제(키워드) DDR , cancer , migration , inviasion
- 발행기관 고려대학교 대학원
- 지도교수 김익영, 양범석
- 발행년도 2009
- 학위수여년월 2009. 8
- 학위명 석사
- 학과 일반대학원 생명공학과
- 세부전공 생화학전공
- 원문페이지 73 p
- 실제URI http://www.dcollection.net/handler/korea/000000009111
- 본문언어 영어
- 제출원본 000045550486
초록/요약
Discoidin domain receptors (DDRs) which consist of DDR1 and DDR2 belong to receptor tyrosine kinase family and become activated by collagens of non-soluble extracellular matrix protein. Although their elevated expressions were reported in various human cancers, more studies are necessary to understand how they contribute to human malignancy. In this study, it was confirmed that DDR1 and DDR2 are often highly expressed in various human cancer cells. A simultaneously elevated expression of both DDR2 and c-Src was observed in highly metastatic cancer cells such as MDA-MB-231 breast cancer cells where DDR2 was constitutively activated and formed a complex with c-Src. After characterizing a novel potent small molecule inhibitor, K110 to inhibit both tyrosine kinases of DDR1 and DDR2 in human cancer cells, we applied this inhibitor to MDA-MB-231 cells. The treatment of K110 inhibited the migration and invasion of MDA-MB-231 cells. This inhibition is correlated with a specific suppression of tyrosine 816 phosohorylation of FAK and reduction of the formation of focal adhesion complex and actin stress fiber. DDR2 was a part of FAK-c-Src complex. Interestingly DDR2 inhibition led to suppression of Akt1 activation. The growth of MDA-MB-231 tumor xenografted in immune-deficient mouse was inhibited by K110. Based on these results we propose that the constitutively activated DDR2 by forming a complex with c-Src contributes to malignant cancer development with promoting cancer cell migration and invasion and this is associated with a cell signaling involving the phosphorylation of tyrosine 861 of FAK and serine 473 of Akt1 and the regulation of focal adhesion complex and actin stress fiber formation.
more목차
Table of Contents i
List of figures iv
List of Table v
Abstract vi
I. Introduction 1
1.1. Discoidin Domain Receptor family 1
1.2. Focal adhesion kinase 3
1.3. Discoidin Domain Receptor 2 and Human diseases 4
1.4. Purpose of this study 7
II. Materials and methods 9
2.1. Antibodies 9
2.2. Cell lines and culture 9
2.3. Generation of stable clonal cell lines 10
2.4. Immunoblotting and immunoprecipitation 11
2.5. Autophosphorylation inhibition assay of cellular DDRs 12
2.6. Wound migration and Matrigel invasion assay 13
2.7. Cellular imaging of focal adhesion complex of cytoskeleton 14
2.8. Xenograft model of tumor growth inhibition 15
III. Results 16
3.1. Various human cancer cell lines often express Discoidin domain receptors at an elevated level 16
3.2. DDR2 is constitutively activated in MDA-MB-231 cells by c-Src kinase and makes a complex with c-Src 20
3.3. K110 is a potent small molecule inhibitor against both of DDR1 and DDR2 tyrosine kinase23
3.4. Inhibition of DDR2 in MDA-MB-231 cells suppresses the collagen dependent migration of MDA-MB-231 cancer cell as well as its invasive ability through matrigel 26
3.5. Inhibition of DDR2 in MDA-MB-231 cells suppresses a specific tyrosine 861 phosphorylation of FAK 30
3.6. Inhibition of DDR2 in MDA-MB-231 cells suppresses the formation of focal adhesion complex and actin stress fiber 33
3.7. Inhibition of DDR2 in MDA-MB-231 cells suppresses activation of Akt1 35
3.8. K110 suppresses the tumor growth of MDA-MB-231 cell in immune deficient mouse xenograft 37
IV. Discussion 39
V. References 48
Abstract in Korean 61
Acknowledgment 63
