Liver metastasis is established by metastasis of micro cell aggregates but not single cells

Kasuya,Kazuhiko; Nagakawa,Yuichi; Hosokawa,Yuichi; Sahara,Yatsuka; Takishita,Chie; Nakajima,Tetsushi; Hijikata,Yosuke; Soya,Ryoko; Katsumata,Kenji; Tsuchida,Akihiko

doi:10.3892/etm.2017.4470

Liver metastasis is established by metastasis of micro cell aggregates but not single cells

Authors:
- Kazuhiko Kasuya
- Yuichi Nagakawa
- Yuichi Hosokawa
- Yatsuka Sahara
- Chie Takishita
- Tetsushi Nakajima
- Yosuke Hijikata
- Ryoko Soya
- Kenji Katsumata
- Akihiko Tsuchida
View Affiliations

Affiliations: Department of Gastrointestinal Surgery, Tokyo Medical University, Shinjuku‑ku, Tokyo 160‑0023, Japan
Published online on: May 18, 2017 https://doi.org/10.3892/etm.2017.4470
Pages: 221-227
Copyright: © Kasuya et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cancer cell engraftment in the target organ is necessary to establish metastasis. Clinically, lymph node metastasis of single cells has been confirmed using cytokeratin staining. In the current study, a LacZ‑labeled cancer cell line was used to visualize intrahepatic metastasis of single cells or liver micrometastasis. KM12SM‑lacZ stably expressing LacZ was prepared with a highly metastatic colon cancer cell line, KM12SM. KM12SM‑lacZ was injected into the spleen of nude mice and following 1 week the spleen was excised. The liver was then examined for metastasis following 1, 2 or 3 weeks. Confirmation of liver metastasis was completed by observing the grade of metastasis. Grade‑1 metastasis (DNA level), human DNA in liver tissue was detected; Grade‑2 metastasis (metastasis of single cells), confirmed by X‑gal staining; Grade‑3 metastasis (histopathological micrometastasis), diagnosed by light microscopy and Grade‑4 metastasis (typical metastasis), easily detected macroscopically or by hematoxylin and eosin staining. The Grade‑1 metastasis detection rates 1, 2 and 3 weeks following splenectomy were 50, 100 and 100%, respectively. Grade‑2 metastasis was not detected by microscopy. The Grade‑3 metastasis detection rates for 1, 2 and 3 weeks were 75, 100 and 100%, respectively. Micrometastasis was observed in the portal vein lumen and wall. The Grade‑4 metastasis detection rates were 50, 100 and 100% for 1, 2 and 3 weeks respectively. Cancer cells were present in vessels surrounding the main tumor. In conclusion, a specific number of cancer cell aggregates may be necessary to establish hematogenous metastasis.

View Figures

View References

1	Nordlinger B, Guiguet M, Vaillant JC, Balladur P, Boudjema K, Bachellier P and Jaeck D: Surgical resection of colorectal carcinoma metastases to the liver. A prognostic scoring system to improve case selection, based on 1568 patients. Association Française de Chirurgie. Cancer. 77:1254–1262. 1996. View Article : Google Scholar : PubMed/NCBI
2	Takahashi M, Hasegawa K, Oba M, Aoki T, Sakamoto Y, Sugawara Y and Kokudo N: Repeat resection leads to long-term survival: Analysis of 10-year follow-up of patients with colorectal liver metastases. Am J Surg. 210:904–910. 2015. View Article : Google Scholar : PubMed/NCBI
3	Mise Y, Imamura H, Hashimoto T, Seyama Y, Aoki T, Hasegawa K, Beck Y, Sugawara Y, Makuuchi M, Nakajima J and Kokudo N: Cohort study of the survival benefit of resection for recurrent hepatic and/or pulmonary metastases after primary hepatectomy for colorectal metastases. Ann Surg. 251:902–909. 2010. View Article : Google Scholar : PubMed/NCBI
4	Löwenthal D, Zeile M, Lim WY, Wybranski C, Fischbach F, Wieners G, Pech M, Kropf S, Ricke J and Dudeck O: Detection and characterisation of focal liver lesions in colorectal carcinoma patients: Comparison of diffusion-weighted and Gd-EOB-DTPA enhanced MR imaging. Eur Radiol. 21:832–840. 2011. View Article : Google Scholar : PubMed/NCBI
5	Ueno H, Mochizuki H, Shirouzu K, Kusumi T, Yamada K, Ikegami M, Kawachi H, Kameoka S, Ohkura Y, Masaki T, et al: Actual status of distribution and prognostic impact of extramural discontinuous cancer spread in colorectal cancer. J Clin Oncol. 29:2550–2556. 2011. View Article : Google Scholar : PubMed/NCBI
6	Fidler IJ: The pathogenesis of cancer metastasis: The ‘seed and soil’ hypothesis revisited. Nat Rev Cancer. 3:453–458. 2003. View Article : Google Scholar : PubMed/NCBI
7	Isaka N, Nozue M, Doy M and Fukao K: Prognostic significance of perirectal lymph node micrometastases in Dukes' B rectal carcinoma: An immunohistochemical study by CAM5.2. Clin Cancer Res. 5:2065–2068. 1999.PubMed/NCBI
8	Okada K, Wada T, Ito K, Takagi Y, Aoki T and Koyanagi Y: Investigation of the anti-tumor activity of hepatic Kupffer cells for control of colon cancer micro-metastasis to the liver via the Fas/Fas ligand system. J Tokyo Med Univ. 61:329–335. 2003.
9	Morikawa K, Walker SM, Nakajima M, Pathak S, Jessup JM and Fidler IJ: Influence of organ environment on the growth, selection, and metastasis of human colon carcinoma cells in nude mice. Cancer Res. 48:6863–6871. 1988.PubMed/NCBI
10	Warburton PE, Greig GM, Haaf T and Willard HF: PCR amplification of chromosome-specific alpha satellite DNA: Definition of centromeric STS markers and polymorphic analysis. Genomics. 11:324–333. 1991. View Article : Google Scholar : PubMed/NCBI
11	Becker M, Nitsche A, Neumann C, Aumann J, Junghahn I and Fichtner I: Sensitive PCR method for the detection and real-time quantification of human cells in xenotransplantation systems. Br J Cancer. 87:1328–1335. 2002. View Article : Google Scholar : PubMed/NCBI
12	Oda H, Ogata Y and Shirouzu K: The effect of angiogenesis inhibitor TNP-470 against postoperative lung metastasis following removal of orthotopic transplanted human colon cancer: An experimental study. Kurume Med J. 48:285–293. 2001. View Article : Google Scholar : PubMed/NCBI
13	Giavazzi R, Jessup JM, Campbell DE, Walker SM and Fidler IJ: Experimental nude mouse model of human colorectal cancer liver metastases. J Natl Cancer Inst. 77:1303–1308. 1986.PubMed/NCBI
14	Fidler IJ: Orthotopic implantation of human colon carcinomas into nude mice provides a valuable model for the biology and therapy of metastasis. Cancer Metastasis Rev. 10:229–243. 1991. View Article : Google Scholar : PubMed/NCBI
15	Takiguchi S, Shimazoe T and Kono A: Antitumor effect of camptothecin analog on liver metastatic model of human colon cancer in nude mice. Gan To Kagaku Ryoho. 21:705–708. 1994.(In Japanese). PubMed/NCBI
16	Chen Y, Zou TN, Wu ZP, Zhou YC, Gu YL, Liu X, Jin CG and Wang XC: Detection of cytokeratin 19, human mammaglobin and carcinoembryonic antigen-positive circulating tumor cells by three-marker reverse transcription-PCR assay and its relation to clinical outcome in early breast cancer. Int J Biol Markers. 25:59–68. 2010.PubMed/NCBI
17	Nomoto S, Nakao A, Ando N, Takeda S, Kasai Y, Inoue S, Kaneko T and Takagi H: Clinical application of K-ras oncogene mutations in pancreatic carcinoma: Detection of micrometastases. Semin Surg Oncol. 15:40–46. 1998. View Article : Google Scholar : PubMed/NCBI
18	Kutun S, Celik A, Cem Kockar M, Erkorkmaz U, Eroğlu A, Cetin A, Erkosar B and Yakicier C: Expression of CK-19 and CEA mRNA in peripheral blood of gastric cancer patients. Exp Oncol. 32:263–268. 2010.PubMed/NCBI
19	Arlt MJ, Banke IJ, Walters DK, Puskas GJ, Steinmann P, Muff R, Born W and Fuchs B: LacZ transgene expression in the subcutaneous Dunn/LM8 osteosarcoma mouse model allows for the identification of micrometastasis. J Orthop Res. 29:938–946. 2011. View Article : Google Scholar : PubMed/NCBI