The establishment and characterization of two new cell lines derived from a single human colonic adenocarcinoma

Verstijnen, C.; Arends, Jan–Willem; Moerkerk, Peter T.M.; Geraedts, J. P. M.; Sekikawa, Koji; Uitendaal, M.P.; Bosman, Fred T.

doi:10.1007/bf02890243

Cited by 16 publications

(5 citation statements)

References 9 publications

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“…Colon carcinoma cell lines with a mucinous phenotype (MC) were Ls174T (ATCC CL‐188) 17 and 5585‐S (kindly provided by Dr PTM Moerkerk, University of Maastricht, The Netherlands). In the latter, signet ring cells were also seen 18. These cells were grown in Dulbecco's modified Eagle's minimum essential medium containing 10% heat‐inactivated fetal calf serum and 0.1% gentamicin.…”

Section: Methodsmentioning

confidence: 99%

Signet ring cell differentiation in mucinous colorectal carcinoma

Börger

Gosens

Jeuken

et al. 2007

The Journal of Pathology

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Approximately 10% of all colorectal carcinomas are mucinous carcinomas, characterized by extracellular mucin. Occasionally, mucin accumulates intracellularly in these tumours, causing signet ring cell differentiation. We hypothesized that signet ring cells arise from a separate genetic pathway. In this study the molecular background of signet ring cell differentiation was investigated by analysing genetic changes, changes in the expression of adhesion molecules, and mucin content. Furthermore, its clinical relevance was addressed. Cell lines of colorectal tumours with non-mucinous (AC), mucinous (MC), and signet ring cell phenotype (MCSRC) were used for Multiplex Ligation-dependent Probe Amplification to detect deletions and amplifications in specific oncogenes and tumour suppressor genes. Furthermore, the expression of E-cadherin, beta-catenin, ITF (intestinal trefoil factor), and MUC2 in signet ring cells was studied by immunohistochemistry, immunofluorescence, and mRNA in situ hybridization. Results were validated using a large cohort of rectal carcinomas from which clinicopathological data were available. Specific amplifications and deletions in cell lines of AC, MC, and MCSRC were detected. Bcl-2 was amplified in MCSRC and MC cell lines, but not in AC cell lines. Bcl-2 FISH analysis confirmed this in patient material. Signet ring cells had decreased expression of adhesion molecules (E-cadherin, beta-catenin) and were strongly positive for ITF and MUC2, two peptides which are normally only produced by goblet cells. RNA in situ hybridization confirmed the production of ITF. Mucinous carcinomas with signet ring cell differentiation presented at a higher T stage than adenocarcinomas and mucinous carcinomas (16% pT4 versus 3-5%, p<0.001) and were more frequently node positive (77% vs 39-44%; p<0.001). Prognosis was significantly worse. In conclusion, the presence of signet ring cells in carcinomas with mucinous differentiation correlates with increased T-stage and poor prognosis. These cells, characterized by ITF and MUC2 production, show disruption of the E-cadherin/beta-catenin complex, as well as amplification of Bcl-2.

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Section: Methodsmentioning

confidence: 99%

Signet ring cell differentiation in mucinous colorectal carcinoma

Börger

Gosens

Jeuken

et al. 2007

The Journal of Pathology

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“…Cell lines derived from non‐dysplastic colonic mucosa: CSC1, CD‐NCM356, CD‐NCM425 and NCM460 have been described previously (Stauffer et al , 1995). Colon carcinoma cell lines with a mucinous phenotype, defined by MUC2 gene expression strongly detectable by Northern blotting, were: LS 174T (ATCC CL‐188), NCI‐H498 (ATCC CCL‐254), 5583‐S and 5583‐E (described by Verstijnen et al ., 1987). Colon carcinoma cell lines with non‐mucinous phenotype: LS 180 (ATCC CL‐187), HT 29 (ATCC HTB‐32), SW403 (ATCC CCL‐230), SW620 (ATCC CCL‐227), COLO 205 (ATCC.…”

Section: Methodsmentioning

confidence: 99%

Differential gene expression in colon carcinoma cells and tissues detected with a cDNA array

et al. 1999

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Expression of selected genes coding for proteins with defined cellular functions was analysed in human cell lines derived from normal colonic mucosa, non‐mucinous colonic carcinomas and mucinous colonic carcinomas. Altered expression of 10 genes in colon carcinoma cells was found by using a cDNA array; 6 of these alterations (60%) were confirmed by Northern blotting or semi‐quantitative reverse transcription‐polymerase chain reaction (RT‐PCR). Among these 6 genes, 3 transcription factors as well as the topoisomerase II α and the mitosis inhibitor WEE1Hu gene were significantly suppressed in the tumour cell lines. In addition, the gene coding for the cell cycle inhibitor p21 was overexpressed only in cell lines derived from mucinous carcinomas. The significant suppression of the kinase WEE1Hu gene in carcinoma cells of both phenotypes and the tendency of the mucinous phenotype to overexpress p21 protein were confirmed in human colon carcinoma tissues. Our data show that the cDNA array method permits a correct identification of changes in gene expression with a relatively high accuracy. The different expression of the p21 gene in the non‐mucinous and mucinous carcinoma cells supports the hypothesis that these phenotypes may develop along different genetic pathways. The detection of WEE1Hu gene suppression in colon carcinoma cells and tissues suggests its potential role in tumourigenesis. Int. J. Cancer 82:868–874, 1999. © 1999 Wiley‐Liss, Inc.

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“…Cell lines derived from the colorectal mucinous carcinomas are limited in number. To our knowledge, only 10 cell lines have been derived from colorectal mucinous carcinoma, but among them only four cell lines (NCI‐H498, 5583‐S, RW‐2982, RW‐7213) possess tumorigenicity in nude mice and maintain abundant mucin production in xenografts 20–22 . With regard to the difficulty of the establishment of cell lines from the mucinous carcinomas, Tibbetts et al .…”

Section: Discussionmentioning

confidence: 99%

“…Mucinous carcinomas of the colorectum have a poor prognosis, and mucin production may be involved in the invasion and metastasis 17–19 . However, the cell lines derived from mucinous carcinomas are quite scarce 20–25 . In the present study, we established a new goblet‐cell differentiated cell line derived from rectal mucinous carcinoma metastasized to the liver and characterized their differentiation and biological properties.…”

mentioning

confidence: 93%

Establishment and characterization of a human colonic mucinous carcinoma cell line with predominant goblet‐cell differentiation from liver metastasis

Yamachika

Nakanishi

Yasui

et al. 2005

Pathology International

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Many human colorectal adenocarcinoma cell lines have been developed. However, differentiated type colorectal cancer cell lines, particularly, the goblet-cell differentiated type, are scarce. In the present study a novel colorectal adenocarcinoma cell line (designated as COLM-6) with predominant goblet-cell differentiation was established from the rectal mucinous adenocarcinoma of a Japanese woman. COLM-6 cells grow in a typical epithelial monolayer in culture. They expressed epidermal growth factor (EGF) receptor and HER2 on their surface and accordingly, their growth was significantly stimulated by EGF, transforming growth factor (TGF)-alpha and heregulin. COLM-6 cells form tumor with typical mucinous adenocarcinomatous appearance in nude mice. Immunohistochemical analysis of these subcutaneous tumors demonstrated that COLM-6 cells strongly express MUC2 as a goblet-cell marker and Cdx2 in the nucleus. Some weakly express villin and carbonic anhydrase 1 as a columnar absorptive-cell marker as well. They were also positive for adenomatous polyposis coli (APC) cytoplasmically and expressed beta-catenin in their cytoplasm and cell membrane without nuclear accumulation. These results indicate that COLM-6 cell line has unique characteristics and may provide a useful tool to study the mechanism of growth and differentiation of colonic epithelium as well as the biological behavior of colorectal mucinous adenocarcinoma.

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The establishment and characterization of two new cell lines derived from a single human colonic adenocarcinoma

Cited by 16 publications

References 9 publications

Signet ring cell differentiation in mucinous colorectal carcinoma

Signet ring cell differentiation in mucinous colorectal carcinoma

Differential gene expression in colon carcinoma cells and tissues detected with a cDNA array

Establishment and characterization of a human colonic mucinous carcinoma cell line with predominant goblet‐cell differentiation from liver metastasis

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