Abstract
Few experimental studies have been conducted to clarify the mechanism of development of metastasis in scirrhous carcinoma of the stomach. In the present study, we attempted to establish gastric carcinoma cell lines by incubation of cancer cells collected from the body fluids of patients with gastric cancer. At the same time, xenografting of these cells to nude mice was performed. It was found that, of the gastric carcinoma cell lines thus established, two cell lines, designated as HSC‐44PE and HSC‐58, formed s.c. tumors with a high infiltrative potential (often invading the lymphatics around the cancer tissue) when implanted. Metastasis to the lymph nodes and lungs was observed in 20–40% of all the animals, indicating that the two cell lines are also capable of metastasizing spontaneously. Through repeated selection, i.e., repeated cycles of removal, culture, and implantation of the HSC cancer cells from metastatic lesions, we obtained 5 subclones of HSC‐44PE and HSC‐58 (designated as m2509, m2615, m2792, m2917, and m2691), which, when implanted orthotopically, exhibited the following characteristics as compared to the parent cells: (1) a higher percentage take (survival), similar frequency of metastasis, shorter time to metastasis (less than 100 days), and consistent metastasizing potential; (2) a relatively high frequency of metastasis to lymph nodes, including distant metastasis to axillary lymph nodes; (3) the potential to cause occasional bloody ascites; (4) enhanced expression of dysadherin, CD44, and other molecules. This is the first report of cultured scirrhous gastric carcinoma cells showing the potential for spontaneous metastasis.
Abbreviations:
- BSA
bovine serum albumin
- CEA
carcinoembryonic antigen
- CDM
chemically defined medium
- FBS
fetal bovine serum
- i.p.
intraperitoneal(ly)
- s.c.
subcutaneous(ly)
- i.v.
intravenous(ly)
- TPA
tissue polypeptide antigen
E‐mail: kyanagih@gan2.res.ncc.go.jp
References
- 1. Fidler IJ. Rationale and methods for the use of nude mice to study the biology and therapy of human cancer metastasis. Cancer Metastasis Rev 1986; 5: 29–49. [DOI] [PubMed] [Google Scholar]
- 2. Fidler IJ. Critical factors in the biology of human cancer metastasis: Twenty‐eighth G. H. A. Clowes Memorial Award Lecture. Cancer Res 1990; 50: 6130–8. [PubMed] [Google Scholar]
- 3. Yanase T, Tamura M, Fujita K, Kodama S, Tanaka K. Inhibitory effect of angiogenesis inhibitor TNP‐470 on tumor growth and metastasis of human cell lines in vitro and in vivo . Cancer Res 1993; 53: 2566–70. [PubMed] [Google Scholar]
- 4. Furukawa T, Fu X, Kubota T, Watanabe M, Kitajima M, Hoffman RM. Nude mouse metastatic models of human stomach cancer constructed using orthotopic implantation of histologically intact tissue. Cancer Res 1993; 53: 1204–8. [PubMed] [Google Scholar]
- 5. Morikawa K, Walker SM, Jessup JM, Fidler IL. In vivo selection of highly metastatic cells from surgical specimens of different primary human colon carcinomas implanted into nude mice. Cancer Res 1998; 48: 1943–8. [PubMed] [Google Scholar]
- 6. Yasoshima T, Denno R, Kawaguchi S, Sato N, Okada Y, Ura H, Kikuchi K, Hirata K. Establishment and characterization of human gastric carcinoma lines with high metastatic potential in the liver: changes in integrin expression associated with the ability to metastasize in the liver of nude mice. Jpn J Cancer Res 1996; 87: 153–60. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7. Li H, Zhang Y‐C, Tsuchihashi Y. Invasion and metastasis of SY86B human gastric carcinoma cells in nude mice. Jpn J Cancer Res 1988; 79: 750–6. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8. Nakanishi H, Yasui K, Yamagata S, Shimizu S, Ando S, Hosoda S. Establishment and characterization of a new spontaneous metastasis model of human gastric carcinoma in nude mice. Jpn J Cancer Res 1991; 82: 927–33. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9. Fujiwara T, Sawada T, Hirakawa‐YS Chung K, Yashiro M, Inoue T, Sowa M. Establishment of lymph node metastatic model for human gastric cancer in nude mice and analysis of factors associated with metastasis. Clin Exp Metastasis 1998; 16: 389–98. [DOI] [PubMed] [Google Scholar]
- 10. Yanagihara K, Seyama T, Tsumuraya M, Kamada N, Yokoro K. Establishment and characterization of human signet ring cell gastric carcinoma cell lines with amplification of the c‐myc oncogene. Cancer Res 1991; 51: 381–6. [PubMed] [Google Scholar]
- 11. Yanagihara K, Kamada N, Tsumuraya T, Amano F. Establishment and characterization of a human gastric scirrhous carcinoma cell line in serum‐free chemically defined medium. Int J Cancer 1993; 54: 200–7. [DOI] [PubMed] [Google Scholar]
- 12. Yanagihara K, Ito A, Toge T, Numoto M. Antiproliferative effects of isoflavones on human cancer cell lines established from the gastrointestinal tract. Cancer Res 1993; 53: 5815–21. [PubMed] [Google Scholar]
- 13. Yanagihara K, Nii M, Tsumuraya M, Numoto M, Seito T, Seyama T. A radiation‐induced murine ovarian granulosa cell tumor line: introduction of v‐ras gene potentiates a high metastatic ability. Jpn J Cancer Res 1995; 86: 347–56. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 14. Ishii Y, Ochiai A, Yamada T, Akimoto S, Yanagihara K, Kitajima M, Hirohashi S. Integrin α6β4 as a suppressor and a predictive marker for peritoneal dissemination in human gastric cancer. Gastroenterology 2000; 118: 497–506. [DOI] [PubMed] [Google Scholar]
- 15. Yanagihara K, Ciardiello F, Talbot N, McGeady ML, Cooper H, Benade L, Salmon DS, Bassin RH. Isolation of a new class of “flat” revertants from ras‐transformed NIH/3T3 cells using cis‐4‐hydroxy‐L‐proline. Oncogene 1990; 5: 1179–86. [PubMed] [Google Scholar]
- 16. Egan SE, Wright JA, Jarolim L, Yanagihara K, Bassin RH, Greenberg AH. Transformation by oncogenes encoding protein kinases induces the metastatic phenotype. Science, 1987; 238: 202–5. [DOI] [PubMed] [Google Scholar]
- 17. Kozaki K, Miyoshi O, Tsukamoto T, Tatematsu Y, Hida T, Takahashi T, Takahashi T. Establishment and characterization of a human lung cancer cell line NCI‐H460‐LNM35 with consistent lymphogenous metastasis via both subcutaneous and orthotopic propagation. Cancer Res 2000; 69: 2535–40. [PubMed] [Google Scholar]
- 18. Yamaguchi K, Ura H, Yasoshima T, Shishido T, Denno R, Hirata K. Establishment and characterization of a human gastric carcinoma cell line that is highly metastatic to lymph nodes. J Exp Clin Cancer Res 2000; 19: 113–20. [PubMed] [Google Scholar]
- 19. 1 Isolation and characterization of highly and rarely metastatic clones from murine colon adenocarcinoma 26. Invasion Metastasis 1984; 4: 84–97. [PubMed] [Google Scholar]
- 20. Takahashi Y, Watanabe M, Mai M, Sasaki T, Fidler IJ. Orthotopic implantation of stomach cancer. Biotherapy 1997; 11: 1011–5. [Google Scholar]
- 21. Yokozaki H, Yasui W, Tahara E. Genetic and epigenetic changes in stomach cancer. Int Rev Cytol 2001; 204: 49–95. [DOI] [PubMed] [Google Scholar]
- 22. Hirohashi S. Inactivation of the E‐cadherin‐mediated cell adhesion system in human cancers. Am. J Pathol 1998; 153: 333–9. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 23. Behrens J, Mareel MM, van Roy FM, Birchmeier W. Dissecting tumor cell invasion: epithelial cells acquire invasive properties after the loss of uvomorulin‐mediated cell‐cell adhesion. J Cell Biol 1989; 108: 2435–447. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 24. Ino Y, Gotoh M, Sakamoto M, Tsukagoshi K, Hirohashi S. Dysadherin, a cancer‐associated cell membrane glycoprotein, down‐regulated E‐cadherin and promotes metastasis. Proc Natl Acad Sci USA 2002; 99: 365–70. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 25. Yokozaki H, Ito R, Nakayama H, Kuniyasu H, Taniyama K, Tahara E. Expression of CD44 abnormal transcripts in human gastric carcinomas. Cancer Lett 1994; 83: 229–34. [DOI] [PubMed] [Google Scholar]
- 26. Tsugawa K, Yonemura Y, Hirono Y, Fushida S, Kaji M, Miwa K, Miyazaki I, Yamamoto H. Amplification of the c‐met, c‐erbB‐2 and epidermal growth factor receptor gene in human gastric cancers: correlation to clinical features. Oncology 1998; 55: 475–81. [DOI] [PubMed] [Google Scholar]
- 27. Kuniyasu H, Yoshida K, Yokozaki H, Yasui W, Ito H, Toge T, Ciardiello F, Persico MG., Saeki T, Salomon DS, Tahara E. Expression of cripto, a novel gene of the epidermal growth factor family, in human gastrointestinal carcinomas. Jpn J Cancer Res 1991; 82: 969–73. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 28. Kuniyasu H, Yasui W, Yokozaki H, Kitadai Y, Tahara E. Aberrant expression of c‐met mRNA in human gastric carcinomas. Int J Cancer 1993; 55: 72–5. [DOI] [PubMed] [Google Scholar]
- 29. Takahashi Y, Cleary KR, Mai M, Kitadai Y, Bucana CD, Ellis LM. Significance of vessel count and vascular endothelial growth factor and its receptor (KDR) in intestinal‐type gastric cancer. Clin Cancer Res 1996; 2: 1679–84. [PubMed] [Google Scholar]
- 30. Huang SP, Wu MS, Wang HP, Yang CS, Kuo ML, Lin JT. Correlation between serum levels of interleukin‐6 and vascular endothelial growth factor in gastric carcinoma. J Gastroenterol Hepatol 2002; 17: 1165–9. [DOI] [PubMed] [Google Scholar]
- 31. Kitadai Y, Haruma K, Sumii K, Yamamoto S, Ue T, Yokozaki H, Yasui W, Ohmoto Y, Kajiyama G, Fidler IJ, Tahara E. Expression of interleukin‐8 correlates with vascularity in human gastric carcinomas. Am J Pathol 1998; 152: 93–100. [PMC free article] [PubMed] [Google Scholar]
- 32. Ueki T, Koji T, Tamiya S, Nakane PK, Tsuneyoshi M. Expression of basic fibroblast growth factor and fibroblast growth factor receptor in advanced gastric carcinoma. J Pathol 1995; 177: 353–61. [DOI] [PubMed] [Google Scholar]
- 33. Nakayama H, Yasui W, Yokozaki H, Tahara E. Reduced expression of nm23 is associated with metastasis of human gastric carcinomas. Jpn J Cancer Res 1993; 84: 184–90. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 34. Fukudome Y, Yanagihara K, Takeichi M, Ito F, Shibamoto S. Characterization of a mutant E‐cadherin protein encoded by a mutant gene frequently seen in diffuse‐type human gastric carcinoma. Int J Cancer 2000; 88: 579–83. [DOI] [PubMed] [Google Scholar]
- 35. Fukuda Y, Kurihara N, Imoto I, Yasui K, Yoshida M, Yanagihara K, Park J‐G, Nakamura Y, Inazawa J. CD44 is a potential target of amplification within the 11p13 amplicon detected in gastric cancer cell lines. Genes Chromosom Cancer 2000; 29: 315–24. [DOI] [PubMed] [Google Scholar]