Intercellular Adhesion Molecule-3 (ICAM-3) Induces Cell Proliferation through the PI3K/Akt Pathway.
- 발행기관 고려대학교 대학원
- 발행년도 2004
- 학위명 박사
- 학과 대학원:의학과 종양생물학 전공
- 식별자(기타) DL:000014914409
- 서지제어번호 000045198392
초록/요약
ICAM3 (Intercellular adhesion molecule-3) is the Ig-like cell adhesion molecule, mostly expresses on human leukocytes and involves in cell-cell interactions. The function as a potent signalling molecule of ICAM3 has not been well understood. Recent studies showed that ICAM3 is involved in maintaining cell survival. So, we examined that the cell proliferation was affected by increased expression of ICAM3 in H1299 lung cancer cell line. We have observed the phosphorylation of FAK, Akt, MAPK pathway through PTEN inhibiton by ICAM3-overexpression, resulting in subsequent leading to down-stream survival signal initiation. We examined the ICAM3-induced cell growth by BrdU incorporation, colony-forming assay, FACS analysis, and cell counting. In addition, we observed that ICAM3 enhanced tumor foci formation by transformation in NIH3T3 cell line, demonstrating that ICAM3 plays a role in tumor proliferation. From the results, we suggest that increased expression of ICAM3 might contributes to tumorigenesis and enhanced cell proliferation through activation of Akt signalling. Therefore, our concepts for ICMA3 could opens new possibilities for the diagnosis, prevention, and treatment of tumors.
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CONTENTS
ABSTRACT I
CONTENTS Ⅲ
LIST OF FIGURES Ⅵ
1. INTRODUCTION 1
1. 1. Ig-like cell adhesion molecule ICAM-3 1
1. 2. Activation of PI3K/Akt pathway 2
1. 3. Phosphorylation of downstream targets through Akt
activation 3
1. 4. Activation of the p44/42 MAP kinase(ERK1/2) 5
1. 5. Tumor suppressor gene PTEN 8
1. 6. LY294002 inhibits the PI3K/Akt pathway 10
1. 7. The PI3K/Akt pathway and transformation 10
2. MATERIALS AND METHODS 13
2. 1. Cell culture and construction of ICAM3 expression
vector 13
2.2. DNA trnasfection for ICAM3 stably cell line and
transient transfections 14
2. 3. Cell growth rate counting 15
2. 4. Measurement of cell proliferation 15
2. 5. Cell cycle analysis and detection of proliferation
fraction 16
2. 6. Colony-forming assay 17
2. 7. Immunoblot analysis for ICAM3 expression at
protein levels 18
2. 8. Inhibitor treatment ; LY294002 and PD98059 19
2. 9. Foci forming assay 20
2. 10. Radiation sensitization of cells 20
3. RESULTS 22
3. 1. An increase of cell proliferation through ICAM3-
overexpression 22
3. 2. ICAM3 induces the PI3K/Akt pathway and activation
of downstream targets 28
3. 3. LY294002 inhibits the PI3K/Akt pathway through
ICAM3-overexpression 32
3. 4. ICAM3-overexpression protects the cells against γ -
ionizing radiation 38
3. 5. ICAM3 is cooperative to malignant transformation
by c-Myc and H-Ras 41
4. DISCUSSION 45
5. REFERENCES 54
국문초록 63
감사의 글 65
LIST OF FIGURES
1. Activation of the PI3K/Akt and ERK/MAPK pathway 6
2. Cellular signaling pathways involved in cancer 7
3. Western blot analysis ; ICAM3 stably transfected into
H1299 23
4. ICAM3 increases cell growth of cancer cell 24
5. ICAM3 expression is associated with the growth of
cancer cell 26
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