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암의 다기능 부착 수용체로서의 syndecan-2
Reports NRF is supported by Research Projects( 암의 다기능 부착 수용체로서의 syndecan-2 | 2004 Year | 박혜인(이화여자대학교) ) data is submitted to the NRF Project Results
Researcher who has been awarded a research grant by Humanities and Social Studies Support Program of NRF has to submit an end product within 6 months(* depend on the form of business)
사업별 신청요강보기
  • Researchers have entered the information directly to the NRF of Korea research support system
Project Number C00024
Year(selected) 2004 Year
the present condition of Project 종료
State of proposition 재단승인
Completion Date 2006년 01월 08일
Year type 결과보고
Year(final report) 2006년
Research Summary
  • Korean
  • Syndecan-2 는 transmembrane heparin sulfate proteoglycan 으로서 다양한 생물학적 기능에 관여한다. 주로 mesenchymal 세포와 여러가지 세포의 이동 환경에서 발현되는 것으로 알려져있으나, 암세포에서의 syndecan-2의 기능은 아직까지 잘 알려져 있지 않다. 이에 본 논문에서는 대장암 세포와 육종암 세포에서의 syndecan-2의 역할과 조절 메커니즘을 살펴보았고, 이를 통해 syndecan-2가 암세포의 암활성을 조절하는 다기능 세포부착 수용체임을 규명하였다.
    우선, syndecan-2가 발암의 여러 단계의 조절에 관여할 수 있기 때문에, 대장암의 초기 단계에서의 syndecan-2의 기능을 알아보기 위하여, 전이성이 낮은 HT-29 세포주에 syndecan-2의 발현을 증가시켰다. HT-29 세포주에 syndecan-2의 증가는 세포와 세포의 경계가 없어지면서 세포가 쌓여자라는 모양의 변형을 유도하였고, 증가된 세포의 이동, 침투, 증식, 그리고 anchorage-independent growth에 기초한 현저히 증가된 암활성을 보였다. 또한, nude mice에 syndecan-2의 증가된 발현을 가진 HT-29 세포주를 주입했을 때, 암의 크기가 증가되었으며, 대장암 환자의 조직을 이용하여 tissue microarray, immunohistochemistry를 실시하였을 때, 대장암 진행의 초기 단계에서 syndecan-2의 발현이 증가됨을 보였다. 이러한 syndecan-2의 발현은 MMP-7 의 발현증가와 밀접하게 연관되어 있으며, syndecan-2에 의해 유도된proMMP-7은 syndecan-2에 결합하여, HT-29 세포주의 세포 표면에 편중되어 위치하였다. 따라서, 이러한 연구 결과는 syndecan-2가 MMP-7의 발현 유도와 그것의 활성을 통하여 HT-29 대장암 세포주의 전이 능력을 증가시킬 수 있음을 보여주는 것이다.
    실제로, 여러 대장암 세포주에서 syndecan-2는 정상세포주에 비해 증가된 발현을 보였다. 재조합된 syndecan-2 의 extracellular 도메인 (2E)을 이용한 adhesion assay나 syndecan-2 항체를 이용한 spreading assay는 syndecan-2가 대부분의 대장암 세포의 부착과 cytoskeletal organization을 조절할 수 있음을 보여주었다. 또한, 이러한 상호작용은 대장암 세포의 증식에 매우 중요함을 보였다. 2E나 syndecan-2 안티센스 cDNA를 이용하여 syndecan-2를 저해했을 때, 세포주기의 G0/G1 arrest 가 유도되었고, 이와는 반대로 p21, p27, 그리고 p53의 발현이 증가되었으며, 대장암 세포의 anchorage-independent growth가 현저히 감소되었다. 따라서, 이러한 결과들은 증가된 syndecan-2의 발현이 대장암 세포의 작용에 매우 중요하며, syndecan-2가 이들 세포들의 세포부착이나 증식의 조절을 통하여 암활성을 조절한다는 것을 제시한다.
    반면에, HT1080 육종암 세포에서 syndecan-2는 암세포의 이동, matrigel로의 침투, 그리고 anchorage-independent growth의 조절에 관여하나, 세포의 ECM의 부착, 증식엔 관여하지 않음으로써, 육종암 세포에서 다른 역할을 수행할 수 있음을 보여주었다. HT1080 세포주에서 증가된 syndecan-2의 발현으로 인한 세포의 이동과 침투의 증가는 focal adhesion kinase (FAK)와 phosphatidylinositol-3 kinase (PI3K) 의 인산화와 서로간의 상호작용의 증가를 유도하였고, Tiam-1의 세포표면으로의 이동과, Rac의 활성을 증가시켰다. 이와는 반대로, 1) 인산화될 수 없는 돌연변이 FAK Y397F 또는 PI3K와의 상호작용을 막는 FAK 돌연변이를 도입시켰을 때, 2) 특이적 PI3K 저해제를 세포에 처리했을 때, 또는 3) siRNA를 이용해 Tiam-1의 발현을 억제시켰을 때, syndecan-2 에 의한 HT1080 세포주의 세포 이동과 침투가 저해되는 것을 관찰하였다. 이러한 결과들은 syndecan-2가 FAK를 통해서 육종암의 암활성을 조절할 수 있음을 보여주는 것이다.
    결론적으로, syndecan-2는 대장암과 육종암 세포 모두의 암활성 조절에 깊게 관여함으로써, 암의 진행에 있어서 다기능 부착 수용체로서 중요한 역할을 수행함을 제시한다.
  • English
  • Syndecan-2 is a member of transmembrane heparan sulfate proteoglycan family in vertebrates and participates in diverse biological processes. In fact, syndecan-2 is mainly expressed in mesenchymal cells and various cells in migratory condition. However, the specific function of syndecan-2 at the cancer cell surface is still unclear. Thus, I examined the roles and regulatory mechanisms of syndecan-2 in high motile colorectal cancer and fibrosarcoma cells and found that syndecan-2 is a multi-functional adhesion receptor to regulate tumorigenic activity of cancer cells.
    Since syndecan-2 might be involved in the regulation of various points during carcinogenesis, its functions were investigated in two different sets of colon cancer cells. To determine syndecan-2 function at the early stage of colon cancer, syndecan-2 was overexpressed in a low metastatic HT-29 cells. Overexpression of syndecan-2 in HT-29 cells induced distinct morphological changes of which piled up in layers without clear boundary and significantly increased tumorigenic activity based on elevated cell migration/invasion, anchorage-independent growth, and proliferation. Consistently, transplantation of HT-29 cells with high expression of syndecan-2 into nude mice showed increased tumor mass, and in tissue microarray and immonohistochemistry of colon cancer patient tissue showed that syndecan-2 was upregulated at early stage in colorectal carcinogenesis. Besides, MMP-7 expression was increased, paralled with syndecan-2 expression. proMMP-7 bound to syndecan-2 and syndecan-2 expression caused polarized MMP-7 localization on cell surface of HT-29 cells. Thus, these results strongly suggest that syndecan-2 enhances the metastatic potential of human colon carcinoma HT-29 cells through the induction and activation of MMP-7.
    In fact, in several colon cancer cell lines, syndecan-2 was highly expressed compared with normal cell lines. Cell biological studies using the extracellular domain of recombinant syndecan-2 (2E) or spreading assay with syndecan-2 antibody-coated plates showed that syndecan-2 mediated adhesion and cytoskeletal organization of most colon cancer cells. This interaction was critical for the proliferation of colon carcinoma cells. Furthermore, inhibition of syndecan-2 expression significantly reduced anchorage-independent growth in colon carcinoma cells. Therefore, increased syndecan-2 expression appears to be a critical for colon carinoma cell behavior, and syndecan-2 regulates tumorigenic activity through regulation of adhesion and proliferation in colon carcinoma cells.
    On the other hand, in HT1080 fibrosarcoma cells, syndecan-2 regulated migration, invasion into Matrigel and anchorage-independent growth, not cell-ECM adhesion or proliferation, suggesting that different functional roles of syndecan-2 in fibrosarcoma cells. Consistent with the increased cell migration/invasion of syndecan-2-overexpressing HT1080 cells, syndecan-2 overexpression increased phosphorylation and interaction of FAK and PI3K, membrane localization of Tiam-1, and activation of Rac. Furthermore, expression of several FAK mutants inhibited syndecan-2-mediated enhancement of anchorage-independent growth in HT1080 cells. Therefore, it is likely that syndecan-2 also regulates the tumorigenic activities of HT1080 fibrosarcoma cells and that tumorigenic activity of syndecan-2 in fibrosarcoma is regulated by FAK.
    Taken together, all these data strongly suggest that syndecan-2 plays critical roles in the neoplastic transformation of normal cells and the regulation of tumorigenic activity of both colon carcinoma and fibrosarcoma cells. Finally, these findings indicate that syndecan-2 plays a critical role as a multi-functional adhesion receptor in cancer progression.
Research result report
  • Abstract
  • Syndecan-2 is a member of transmembrane heparan sulfate proteoglycan family in vertebrates and participates in diverse biological processes. In fact, syndecan-2 is mainly expressed in mesenchymal cells and various cells in migratory condition. However, the specific function of syndecan-2 at the cancer cell surface is still unclear. Thus, I examined the roles and regulatory mechanisms of syndecan-2 in high motile colorectal cancer and fibrosarcoma cells and found that syndecan-2 is a multi-functional adhesion receptor to regulate tumorigenic activity of cancer cells.
    Since syndecan-2 might be involved in the regulation of various points during carcinogenesis, its functions were investigated in two different sets of colon cancer cells. To determine syndecan-2 function at the early stage of colon cancer, syndecan-2 was overexpressed in a low metastatic HT-29 cells. Overexpression of syndecan-2 in HT-29 cells induced distinct morphological changes of which piled up in layers without clear boundary and significantly increased tumorigenic activity based on elevated cell migration/invasion, anchorage-independent growth, and proliferation. Consistently, transplantation of HT-29 cells with high expression of syndecan-2 into nude mice showed increased tumor mass, and in tissue microarray and immonohistochemistry of colon cancer patient tissue showed that syndecan-2 was upregulated at early stage in colorectal carcinogenesis. Besides, MMP-7 expression was increased, paralled with syndecan-2 expression. proMMP-7 bound to syndecan-2 and syndecan-2 expression caused polarized MMP-7 localization on cell surface of HT-29 cells. Thus, these results strongly suggest that syndecan-2 enhances the metastatic potential of human colon carcinoma HT-29 cells through the induction and activation of MMP-7.
    In fact, in several colon cancer cell lines, syndecan-2 was highly expressed compared with normal cell lines. Cell biological studies using the extracellular domain of recombinant syndecan-2 (2E) or spreading assay with syndecan-2 antibody-coated plates showed that syndecan-2 mediated adhesion and cytoskeletal organization of most colon cancer cells. This interaction was critical for the proliferation of colon carcinoma cells. Furthermore, inhibition of syndecan-2 expression significantly reduced anchorage-independent growth in colon carcinoma cells. Therefore, increased syndecan-2 expression appears to be a critical for colon carinoma cell behavior, and syndecan-2 regulates tumorigenic activity through regulation of adhesion and proliferation in colon carcinoma cells.
    On the other hand, in HT1080 fibrosarcoma cells, syndecan-2 regulated migration, invasion into Matrigel and anchorage-independent growth, not cell-ECM adhesion or proliferation, suggesting that different functional roles of syndecan-2 in fibrosarcoma cells. Consistent with the increased cell migration/invasion of syndecan-2-overexpressing HT1080 cells, syndecan-2 overexpression increased phosphorylation and interaction of FAK and PI3K, membrane localization of Tiam-1, and activation of Rac. Furthermore, expression of several FAK mutants inhibited syndecan-2-mediated enhancement of anchorage-independent growth in HT1080 cells. Therefore, it is likely that syndecan-2 also regulates the tumorigenic activities of HT1080 fibrosarcoma cells and that tumorigenic activity of syndecan-2 in fibrosarcoma is regulated by FAK.
    Taken together, all these data strongly suggest that syndecan-2 plays critical roles in the neoplastic transformation of normal cells and the regulation of tumorigenic activity of both colon carcinoma and fibrosarcoma cells. Finally, these findings indicate that syndecan-2 plays a critical role as a multi-functional adhesion receptor in cancer progression.
  • Research result and Utilization method
  • 세포 표면 부착 수용체인 syndecan-2는 대장암 진행의 초기 단계에 증가하여 대장암 세포들의 부착과 증식의 조절을 통해 암활성을 증가시켰고, FAK-PI3K 신호전달 체계를 통해서 HT1080 육종암 세포주의 migration/invasion의 증가에 기초한 암활성에 중요한 역할을 하는것을 알수 있었다. 또한 육종암세포에서 syndecan-2는 세포의 ECM의 부착, 증식엔 관여하지 않음으로써 대장암 세포와는 다른 작용을 보이는 것을 확인 할 수 있었다. 따라서, 이러한 결과는 syndecan-2가 암세포의 암활성을 조절하는 다기능 세포부착 수용체임을 규명해주는 것이며, 이러한 조직 특이적 조절 기작을 이용하여 대장암 세포와 육종암 세포의 암활성을 억제시킬 수 있는 억제제 개발에 이 결과를 이용할 수 있을것으로 생각된다.
  • Index terms
  • Syndecan-2, Migration/Invasion, Fibrosarcoma, colon cancer, tumorigenic activity.
  • List of digital content of this reports
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