폐암의 림프절 전이 및 신생림프관형성에 있어서 Shh/Gli 신호전달 경로의 역할 규명
The crucial role analysis of Shh/Gli signaling pathway in lymph node metastasis and lymphangiogenesis of lung cancer
- 주제(키워드) Lung cancer , Shh/Gli signaling pathway , Lymphangiogenesis
- 발행기관 고려대학교 대학원
- 지도교수 김학제
- 발행년도 2011
- 학위수여년월 2011. 2
- 학위구분 박사
- 학과 일반대학원 의학과
- 세부전공 흉부외과학 전공
- 원문페이지 47 p
- 실제URI http://www.dcollection.net/handler/korea/000000024564
- 본문언어 한국어
- 제출원본 000045635187
초록/요약
Background: The Shh/Gli signaling pathway (Sonic hedgehog/Glioma-associated oncogene homolog) plays a critical role in the differentiation and tumorigenesis in human cells. Some studies report that this pathway can induce lymphangiogenesis which is known to be related to lymph node metastasis. In non-small cell lung cancer (NSCLC), however, there have been no studies regarding the role of the Shh/Gli signaling pathway in lymphangiogenesis and lymph node metastasis. Therefore, in this study, we investigated whether the activation of the Shh/Gli signaling pathway could affect lymph node metastasis and lymphangiogenesis in NSCLC. Methods: To elucidate the effects of the Shh/Gli signaling pathway on proliferation, migration, invasion and lymphangiogenesis in NSCLC cells, we conducted MTT, wound healing migration, Matrigel invasion, RT-PCR of lymphangiogenic factor (VEGF-D and LYVE-1),and in vitro lymphangiogenesis using Human MicroVascular endothelial cells (HMVEC) with or without N-Shh peptide (recombinant Shh), KAAD-cyclopamine, (Shh/Gli pathway inhibitor) and Glis si RNA (Knockdown of Gli-1,2,3 genes). To investigate the correlation among the Shh/Gli signaling pathway, lymph node metastasis and lymphangiogenesis in NSCLC, we performed immunohistochemical assays for the biomarkers of Shh/Gli signaling pathway (Shh and Gli) and lymphangiogenesis (VEGF-D and LYVE-1) with primary tumor tissues, metastatic lymph nodes, and non-metastatic lymph nodes in 42 NSCLC patients. In addition, we analyzed the statistical differences in the expression of each biomarker according to clinicopathological data, including age, gender, tumor size, lymph node metastasis status and smoking habit. Results: We found that the N-Shh peptide enhanced the migration/invasion of NSCLC cells, whereas no increase was observed in cells that were treated with KAAD-cyclopamine or Glis siRNA in wound healing migration and Matrigel invasion assays. However, in MTT assays there was no change in the proliferation of cells for 72 hours after the same treatment. Also, we observed in NSCLC cells that the N-Shh peptide could induce the expression of VEGF-D and LYVE-1, whereas KAAD-cyclopamine or Glis si RNA reduced the expression of those, according to the results of RT-PCR. In the microscopic finding of in vitro lymphangiogenesis, newly developed lymphatic tubules were detected after treatment of N-Shh peptide, while suppressed formation of lymphatic tubules were observed after treatment of KAAD-cyclopamine. Immunohistochemical staining of NSCLC tissues revealed that the expression of Shh, Gli, VEGF-D, and LYVE-1 were elevated in the primary tumors of NSCLC and metastatic lymph nodes, compared with those of non-metastatic lymph nodes. In contrast, there was no significant difference in the expression of biomarkers according to each clinicopathological data. Conclusion: The Shh/Gli signaling pathway could be related to the motility/invasiveness of NSCLC cells and according to these results we further suggest that the Shh/Gli signaling pathway may induce lymph node metastasis in tumor cells. In addition, the signaling pathway could be associated with lymphangiogenesis. Taken together, we suggest that the Shh/Gli signaling pathway could be a target of treatment by inhibiting lymph node metastasis and lymphangiogenesis in NSCLC.
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영문요약 i
I. 서론------------------------------1
II. 연구재료와 실험 방법-------------- 3
III. 결과----------------------------- 12
IV. 고찰----------------------------- 15
V. 결론----------------------------- 21
VI. 참고 문헌------------------------ 22
표 및 그림-----------------------------------26
