臺灣博碩士論文加值系統

卵巢癌是目前最致命的婦科惡性腫瘤，癌細胞轉移是最主要的致命病因；包含卵巢癌，癌細胞的特定轉移模式被稱為「器官特異性」。
此論文中，我們使用了CB17/lcr-PRkdc scid/Crl免疫缺陷小鼠，檢視不同卵巢癌細胞在小鼠體內的器官特異性。原發腫瘤取得的A1847和A2780卵巢癌細胞株經由尾靜脈注射，會趨向卵巢轉移；而從腹水取得的Skov3卵巢癌細胞株經由尾靜脈注射，則轉移至肺臟。
藉由分析癌細胞的基本特性包含型態、移動能力、侵入能力以及增殖能力，試著去解釋不同卵巢癌細胞株的器官特異性。經由小鼠尾靜脈注射後48鐘頭，我們觀察到A1847和A2780細胞轉移至卵巢，而Skov3細胞在相同條件下則轉移至肺臟。將A1847細胞以及A2780細胞直接注射植入肺臟，細胞會轉移至卵巢；Skov3細胞原位打入卵巢，則會轉移至肺臟。
有報導指出，乳癌循環腫瘤細胞會腫瘤自播(tumor self-seeding)，我們觀察到卵巢癌Skov3細胞也有相似的現象。臨床研究顯示，兩側卵巢均有惡性腫瘤的案例有25%；我們也發現Skov3細胞能藉轉移到另外一側的卵巢，並且創傷癒合會促進此現象。
此論文說明了不同卵巢癌細胞株的特殊器官特異性，以及證實了在癌細胞轉移機制的限制步驟中，成功定殖(colonization)且生長較早期癌細胞附著於器官組織更為重要。並且也證明創傷癒療會促進細胞定殖。

Ovarian cancer is the most lethal gynecologic malignancy, and the majority of deaths occur due to metastatic disease. Many studies have identified the specific pattern of metastasis, termed organ tropism, including ovarian cancer.
In this thesis, we used the CB17/lcr-PRkdc scid/Crl mice to show that different ovarian cancer cell lines had different organ tropism. The A1847 line and A2780 line derived from primary ovarian tumors displayed ovary tropism upon intravenous injection. The Skov3 cells derived from the ascites of ovarian adenocarcinoma form metastases in lung upon intravenous injection. We analyzed the properties of ovarian cancer cells including morphology, migration ability, invasion ability, and proliferation in order to explain the different organ tropism. And we observed A1847 and A2780 cells metastasized to ovary 48 hours after intravenous injection, whereas Skov3 cells metastasized to lung tissue under the similar condition. When A1847 and A2780 cells were implanted directly into lung, they metastasized to ovary. Skov3 cells which were implanted orthotopically into ovary metastasized to lung.
It was shown that in breast cancer, circulating tumor cells self-seeded their tumor origin, we observed similar phenomenon in Skov3 cells. Accumulated clinical studies revealed 25% ovarian cancer cases involved bilateral ovarian carcinomas. We found that Skov3 cells were able to metastasize to contralateral ovary, and this was promoted by wound healing.
This thesis demonstrates the distinctive organ tropism of ovarian cancer cell lines, and the limiting step of metastasis resides more in successful colonization than initial targeting of the organs. Wounding healing could promote cancer cell colonization.

中文摘要 i
Abstract ii
Table of Contents iv
List of figures vi
Figures 41 vi
Chapter 1: Introduction and literature review 1
1.1. Ovarian cancer 1
1.2. Metastasis of cancer cells 2
1.3. Current knowledge about the molecular mechanisms of ovarian cancer metastasis 5
1.4. Tumor Seeding by Circulating Cancer Cells 6
1.5. Specific aim and impact of this thesis 7
Chapter 2: Material and Methods 8
2.1. Cell lines 8
2.2. Cell migration/invasion assay and in vitro selection 8
2.3. MTS cell proliferation assay 9
2.5. Blood perfusion in lung 10
2.6. Orthotopic and lung implantation xenograft mouse model 10
2.7. Quantitate the human cancer cells in the tissue 11
2.8. Quantitative real-time PCR for genes 11
2.9. Statistical analysis 12
2.10. Human Cytokine Antibody array 12
Chaper 3: Result 13
3.1. The cell morphology of ovarian cancer cell lines. 13
3.2. The migration and invasive abilities of Skov3 cells were significantly higher than A1847 and A2780 cells. 13
3.3. The proliferation rates were not dramatically different in invasive sublines parental lines. 14
3.4. A1847 and A2780 cells were ovary tropic, whereas Skov3 cells were lung tropic. 14
3.5. Ovarian epithelial cancer cell line, A2780 cells and its invasive subline could extravasate to lung tissue whereas A1847 cells and its invasive subline could not, but all four lines could metastasize to ovary. 15
3.6. Skov3 ovarian cancer cells derived from ascites can extravasate into lung and metastasize to ovary. 17
3.7. Skov3 cells and its invasive subline Skov3-I6 cells colonized not only in lung but also in ovary three weeks post-intravenous injection. 18
3.8. Skov3 cells and its invasive subline Skov3-I6, metastasize to lung when orthotopically implanted into ovary 19
3.9. A1847, A2780, and their invasive sublines which were implanted directly into lung could colonize in lung and metastasize to ovary. 20
3.10. Disseminated tumor cells from one side of ovary could metastasize to contralateral ovary. 21
3.11. Wound healing may attract the disseminated ovarian cancers cells 22
Chaper 4: Discussion 24
4.1. Organ tropism in ovarian cancer cell lines 24
4.2. The invasive abilities of ovarian cancer cells and cytokines released by them may influence the extravasation to lung tissue. 26
4.3. Extravasation to specific tissue was one of the determinants of organ tropism. 27
4.4. The microenvironment at the metastatic site is an important factor determining organ tropism. 28
4.5. The metastatic route of ovarian cancer cells may affect the efficiency of metastasis. 30
4.6. Wound healing promote tumor self-seeding and seeding of circulating tumor cells. 32
4.7. Conclusion 34
Reference 35
Table 39
Table 1. The incidence of tumor in lung or ovary through tail vein injection. 39
Table 2. The incidence of tumor formation in ovary when implanted into lung. 40
Figures 41
Figure 1. The cell morphology in regular culture condition before and after in vitro Boyden chamber invasion selection. 41
Figure 2. Compare migration and invasion ability among ovarian cancer cells and their invasive sublines. 42
Figure 3. MTS proliferation assay. 43
Figure 4. The organ tropism of ovarian cancer cells. 44
Figure 5. A1847 and A1847-I4 cells did not metastasize into lung but could metastasize to ovary 24 or 48 hours post-intravenous-injected. 45
Figure 6. A2780 and A2780-I4 cells metastasized partially to lung and ovary 24 or 48 hours post-intravenous-injection. 46
Figure 7. Skov3 and Skov3-I6 cells metastasized to lung and partially to ovary 48 hours post-intravenous-injection. 47
Figure 8. Skov3 and Skov3-I6 cells colonized in lung and metastasized back to ovary. 48
Figure 9. Skov3 and Skov3-I6 cells implanted orthotopically into ovary metastasized to lung. 49
Figure 10. A1847, A2780, and their invasive sublines which were implanted into lung could colonize in lung and metastasize to ovary. 51
Figure 11. Preferential contralateral ovary seeding by disseminated Skov3 and Skov3-I6 cell. 53
Figure 12. Disseminated Skov3 and Skov3-I6 cancer cells metastasized to contralateral ovary may be influenced by wound healing. 54
Appendix 55
Appendix 1. 55

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