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Correlation of cellular differentiation in human colorectal carcinoma and adenoma cell lines with metabolite profiles determined by 1H magnetic resonance spectroscopy
Wanda B. MaCkinnon,
Lily Huschtscha,
Kathy Dent,
Rebecca Hancock,
Christos Paraskeva,
Carolyn E. Mountford,
Corresponding Author
Wanda B. MaCkinnon
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia. Fax: 61 2 692 4105Search for more papers by this authorLily Huschtscha
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia
Search for more papers by this authorKathy Dent
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia
Search for more papers by this authorRebecca Hancock
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia
Search for more papers by this authorChristos Paraskeva
Department of Pathology, University of Bristol, Bristol BS81TD, UK
Search for more papers by this authorCarolyn E. Mountford
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia
Search for more papers by this authorWanda B. MaCkinnon,
Lily Huschtscha,
Kathy Dent,
Rebecca Hancock,
Christos Paraskeva,
Carolyn E. Mountford,
Corresponding Author
Wanda B. MaCkinnon
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia. Fax: 61 2 692 4105Search for more papers by this authorLily Huschtscha
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia
Search for more papers by this authorKathy Dent
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia
Search for more papers by this authorRebecca Hancock
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia
Search for more papers by this authorChristos Paraskeva
Department of Pathology, University of Bristol, Bristol BS81TD, UK
Search for more papers by this authorCarolyn E. Mountford
Membrane-MR Unit, Cancer Medicine, University of Sydney, Sydney, N.S.W. 2006, Australia
Search for more papers by this authorAbstract
The aim was to determine whether proton magnetic resonance spectroscopy (MRS) could grade human colorectal cells of differing malignant potential. A cell model of tumour development and progression comprising 2 non-tumorigenic adenoma lines and 4 carcinoma lines of increasing tumorigenicity was chosen. A gradual reduction in cellular differentiation and an accumulation of genetic alterations from adenoma to carcinoma characterized the selected cell lines. One-dimensional and 2-dimensional MRS showed that reduced differentiation in the cell model correlated with an increase in the levels of lipid, metabolites, the glycosylation intermediate uridine diphospho-N-acetylglucosamine and cell-surface fucosylation. Mutations involving the K-ras, APC and DCC genes are present both in adenoma- and in carcinoma-derived lines in this model, but the first evidence of an abnormality in the p53 gene was concomitant with the cells' ability to grow as a tumour in athymic nude mice. This genetic change coincided with the detection, by MRS, of UDP-hexose (ribose moiety, 2D MRS cross peak between H2 at 4.38 ppm and HI at 5.99 ppm) and the appearance of an additional fucosyl resonance (cross peak between-CH3 at 1.41 and H5 at 4.30 ppm) in the least tumori-genic of the carcinoma cell lines. An increase in complexity of the fucosylation spectral pattern was observed with further cellular de-differentiation and increased tumorigenicity. Collectively these data support the existence of an adenoma-carcinoma sequence.
References
- Abe, K., Hakomori, S.-I. and Oshiba, S., Differential expression of difucosyl type-2 chain (Ley) defined by monoclonal antibody AH6 in different locations of colonic epithelia, various histological types of colonic polyps, and adenocarcinomas. Cancer Res., 46, 2639–2644 (1986).
-
Atkinson, P. H. and
Hakimi, J.,
Alterations in glycoproteins of the cell surface.
In:
W. J. Lennarz (ed.),
The biochemistry of glycoproteins and proteoglycans, Chapter 5,
pp. 191–239,
Plenum Press, New York
(1980).
10.1007/978-1-4684-1006-8_5 Google Scholar
- Baker, S. J., Preisinger, A. C., Jessup, J. M., Paraskeva, C., Markowitz, S., Willson, J. K. V., Hamilton, S. and Vogelstein, B., p53 gene mutations occur in combination with 17p allelic deletions as late events in colorectal tumorigenesis. Cancer Res., 50, 7717–7722 (1990).
- Bloom, M., Holmes, K. T., Mountford, C. E. and Williams, P. G., Complete proton magnetic resonance in whole cells. J. magn. Reson., 69, 73–91 (1986).
- Bos, J. L., Fearon, E. R., Hamilton, S. R., Verlaan de Vries, M., Van Boom, J. H., Van der Eb, A. J. and Vogelstein, B., Prevalence of ras gene mutations in human colorectal cancers. Nature (Lond.), 327, 293–297 (1987).
- Callies, R., Sri-Pathmanathan, R. M., Ferguson, D. Y. P. and Brindle, K. M., The appearance of neutral lipid signals in the 1H NMR spectra of a myeloma cell line correlates with the induced formation of cytoplasmic lipid droplets. Magn. Reson. Med., 29, 546–550 (1993).
- Catalogue of Cell Lines and Hybridomas. In: R. Hay, M. Macy, T. R. Chen, P. McClintock and Y. Reid (eds.), American Type Culture Collection, 6th ed., Maryland (1988).
- Delikatny, E. J., Hull, W. E. and Mountford, C. E., The effect of altering time domains and window functions in two-dimensional proton COSY spectra of biological specimens. J. magn. Res., 94, 563–573 (1991).
- Delikatny, E. J., Russell, P., Hunter, J. C., Hancock, R., Atkinson, K., van Haaften-Day, C. and Mountford, C. E., Proton MR and human cervical neoplasia. I. Ex vivo spectroscopy allows distinction of invasive carcinoma of the cervix from carcinoma in situ and other pre-invasive lesions. Radiology, 188, 791–796 (1993).
- Ende, D. A., Lean, C. L., Mackinnon, W. B., Chapuis, P., Newland, R. C., Russell, P., Bokey, E. L. and Mountford, C. E., Human colorectal adenoma-carcinoma sequence documented by 1H MRS (ex vivo). Proc. Soc. magn. Reson. Med., 2, 1033 (1993).
- Farr, C. J., Marshall, C. J., Easty, D. J., Wright, N. A., Powell, S. C. and Paraskeva, C., A study of ras gene mutations in colonic adenomas from familial polyposis coli patients. Oncogen, 3, 673–678 (1988).
- Fearon, E. R., Cho, K. R., Nigro, J. M., Kern, S. E., Simons, J. W., Ruppert, J. M., Hamilton, S. R., Preisinger, A. C., Thomas, G., Kinzler, K. W. and Vogelstein, B., Identification of a chromosome 18q gene that is altered in colorectal cancers. Science, 247, 49–56 (1990).
- Fearon, E. R. and Vogelstein, B., A genetic model for colorectal tumorigenesis. Cell, 61, 759–767 (1990).
- Galons, J. P., Fantini, J., Vion-Dury, J., Cozzone, P. and Canioni, P., Metabolic changes in undifferentiated and differentiated human colon adenocarcinoma cells studied by multinuclear magnetic resonance spectroscopy. Biochimie, 71, 949–961 (1989).
- Goyette, M. C., Cho, K., Fasching, C. L., Levy, D. B., Kinzler, K. W., Paraskeva, C., Vogelstein, B. and Stanbridge, E. J., Progression of colorectal cancer is associated with multiple tumor-suppressor-gene defects but inhibition of tumorigenicity is accomplished by correction of any single defect via chromosome transfer. Mol. cell. Biol., 12, 1387–1395 (1992).
- Hakomori, S., Aberrant glycosylation in tumours and tumour-associated carbohydrate antigens. Advanc. Cancer Res., 52, 257–331 (1989).
- Hill, M. J., From adenoma to carcinoma of the colorectum. In: W. J. Eylenbosch, M. Kirsch-Valders, A. Deleener and W. Weyler (eds.), Recent results in cancer research, Vol. 122, pp. 71–84, Springer, Heidelberg (1991).
- Holmes, K. T. and Mountford, C. E., Identification of triglyceride in malignant cell membranes. J. mag. Res., 93, 407–409 (1991).
- Huschtscha, L., Rozengurt, E. and Bodmer, W., Growth factor requirements of human colorectal tumour cells: relations to cellular differentiation. Europ. J. Cancer, 27, 1680–1684 (1991).
- Jesson, J. P., Meakin, P. and Kneissel, G., Homonuclear decoupling and peak elimination of Fourier transform nuclear magnetic resonance. J. Amer. chem. Soc., 95, 618–620 (1973).
- Kim, Y. S., Yuan, M., Itzkowitz, S. H., Sun, Q., Kaizu, T., Palekar, A., Trump, B. F. and Hakomori, S., Expression of Ley and extended Ley blood-group-related antigens in human malignant, pre-malignant, and non-malignant colonic tissue. Cancer Res., 46, 5985–5992 (1986).
- Kinzler, K. W. and 20 others, Identification of FAP locus genes from chromosome 5q21. Science, 253, 661–665 (1991).
- Lean, C. L., Mackinnon, W. B., Delikatny, E. J., Whitehead, R. H. and Mountford, C. E., Cell-surface fucosylation and magnetic resonance spectroscopy characterization of human malignant colorectal cells. Biochemistry, 31, 11095–11105 (1992).
- Lean, C. L., Mackinnon, W. B. and Mountford, C. E., Fucose in 1H COSY spectra of plasma membrane fragments shed from human malignant colorectal cells. Magn. Reson. Med., 20, 306–311 (1991).
- Lean, C. L., Newland, R. C., Ende, D. A., Bokey, E. L., Smith, I. C. P. and Mountford, C. E., Assessment of human colorectal biopsies by 1H MRS: correlation with histopathology. Magn. Reson. Med., 30, 525–533 (1993).
- Leibovitz, A., Stinson, J. C., McCoombs III, W. B., McCoy, C. E., Mazur, K. C. and Mabry, N. D., Classification of human colorectal adenocarcinoma cell lines. Cancer Res., 36, 4562–4569 (1976).
- Mackinnon, W. B., May, G. L. and Mountford, C. E., Esterified cholesterol and triglyceride are present in plasma membranes of Chinese-hamster ovary cells. Europ. J. Biochem., 205, 827–839 (1992).
- Manning, A. M., Williams, A. C., Game, S. M. and Paraskeva, C., Differential sensitivity of human colonic adenoma and carcinoma cells to transforming growth factor β(TGF-β): conversion of an adenoma cell line to a tumorigenic phenotype is accompanied by a reduced response to the inhibitory effects of TGF-β. Oncogene, 6, 1471–1476 (1991).
- May, G. L., Wright, L. C., Holmes, K. T., Williams, P. G., Smith, I. C. P., Wright, P. E., Fox, R. M. and Mountford, C. E., Assignment of methylene proton resonances in NMR spectra of embryonic and transformed cells to plasma membrane triglyceride. J. biol. Chem., 261, 3048–3053 (1986).
- May, G. L., Wright, L. C., Lean, C. L. and Mountford, C. E., Identification of 1–0-alkyl-1–2, 3-diacyl-sn-glycerol in plasma membranes of cancer cells. J. mag. Res., 98, 622–627 (1992).
- Mori, M., Shimono, R., Adachi, Y., Matsuda, H., Kuwano, H., Sugimachi, K., Ikeda, M. and Saku, M., Transitional mucosa in human colorectal lesions. Dis. Colon Rectum, 33, 498–501 (1990).
- Mountford, C. E., Lean, C., Mackinnon, W. B. and Russell, P., Proton magnetic resonance in pathology. In: G. A. Webb (ed.), Annual reports on NMR spectroscopy, Vol. 27, pp. 173–215, Academic Press, London (1993).
- Mountford, C. E. and Wright, L. C., Organization of lipids in the plasma membranes of malignant and stimulated cells: a new model. TIBS, 13, 172–177 (1988).
- Nishicho, I. and 21 others, Mutations of chromosome 5q21 genes in FAP and colorectal-cancer patients. Science, 253, 665–669 (1991).
- Paraskeva, C., Buckle, B. G., Sheer, D. and Wigley, C. B., The isolation and characterization of colorectal epithelial cell lines at different stages in malignant transformation from familial polyposis coli patients. Int. J. Cancer, 34, 49–56 (1984).
- Paraskeva, C., Corfield, A. P., Harper, S., Hague, A., Audcent, K. and Williams, A. C., Colorectal carcinogenesis: sequential steps in the in vitro immortalization and transformation of human colonic epithelial cells (review). Anticancer Res., 10, 1189–1200 (1990).
- Paraskeva, C., Finerty, S. and Powell, S., Immortalization of a human colorectal adenoma cell line by continuous in vitro passage: possible involvement of chromosome 1 in tumour progression. Int. J. Cancer, 41, 908–912 (1988).
- Pignatelli, M. and Bodmer, W. F., Genetics and biochemistry of collagen-binding-triggered glandular differentiation in a human colon carcinoma cell line. Proc. nat. Acad. Sci. (Wash.), 85, 5561–5565 (1988).
- Richart, R. M., Fu, Y. S. and Winkler, B., Pathology of cervical squamous and glandular intraepithelial neoplasia. In: M. Coppleson, J. M. Monaghan, C. P. Morrow and M. H. N. Tattersall (eds.), Gynecologic oncology: fundamental principles and clinical practice, 2nd ed., Part V, pp. 557–570, Churchill Livingstone, Edinburgh (1992).
- Richman, P. I. and Bodmer, W. F., Monoclonal antibodies to human colorectal epithelium: markers for differentiation and tumour characterization. Int. J. Cancer, 39, 317–328 (1987).
- Russell, P., Lean, C. L., Delbridge, L., Wong, G., Dowd, S. and Mountford, C. E., Proton magnetic resonance and human thyroid neoplasms. I: Ex vivo diagnosis of potentially metastatic follicular tumours by magnetic resonance spectroscopy. Amer. J. Med., 96, 383–388 (1994).
- Rutzky, L. P., The biology of human colon tumor cells in culture. Advanc. Cell Culture, 4, 47–83 (1985).
- Shedd, S. F., Lutz, N. W. and Hull, W. E., The influence of medium formulation on phosphomonoester and UDP-hexose levels in cultured human colon tumor cells as observed by 31P NMR spectroscopy. NMR Biomed., 6, 254–263 (1993).
- Smith, I. C. P., Princz, E. J. and Saunders, J. K., Magnetic resonance spectroscopy in cancer research. J. Canad. Ass. Radiol., 41, 32 (1990).
- Smith, K. J., Johnson, K. A., Bryan, T. M., Hill, D. E., Markowitz, S., Willson, J. K. V., Paraskeva, C., Petersen, G. M., Hamilton, S. R., Vogelstein, B. and Kinzler, K. W., The APC gene product in normal and tumor cells. Proc. nat. Acad. Sci., 90, 2846–2850 (1993).
- Sze, D. Y. and Jardetzky, O., Determination of metabolite and nucleotide concentrations in proliferating lymphocytes by 1H-NMR of acid extracts. Biochim. biophys. Acta, 1054, 181–197 (1990a).
- Sze, D. Y. and Jardetzky, O., Characterisation of lipid composition in stimulated human lymphocytes by 1H-NMR. Biochim. biophys. Acta, 1054, 198–206 (1990b).
- Whitehead, R. H., Jones, J. K., Gabriel, A. and Lukeis, R. E., A new colon-carcinoma cell line (LIM1863) that grows as organoids with spontaneous differentiation into crypt-like structures in vitro. Cancer Res., 47, 2683–2689 (1987).
- Whitehead, R. H., Macrae, F. A., St. John, D. J. B. and Ma, J., A colon-cancer cell line (LIM1215) derived from a patient with inherited non-polyposis colorectal cancer. J. nat. Cancer Inst., 74, 759–765 (1985).
- Wice, B. M., Trugnan, G., Pinto, M., Rousset, M., Chevalier, G., Dussaulx, E., Lacroix, B. and Zweibaum, A., The intracellular accumulation of UDP-N-acetylhexosamines is concomitant with the inability of human colon cancer cells to differentiate. J. biol. Chem., 260, 139–146 (1985).
- Williams, A. C., Browne, S. J., Yeudal, W. A., Paterson, I. C., Marshall, C. J., Lane, D. P. and Paraskeva, C., Molecular events including p53 and k-ras alterations in the in vitro progression of a human colorectal-adenoma cell line to an adenocarcinoma. Oncogene, 8, 3063–3072 (1993).
- Williams, A. C., Harper, S. J. and Paraskeva, C., Neoplastic transformation of a human colonic epithelial cell line: in vitro evidence for the adenoma-to-carcinoma sequence. Cancer Res., 50, 4724–4730 (1990).
- Willson, J. K. V., Bittner, G. N., Oberley, T. D., Meisner, L. F. and Weese, J. L., Cell culture of human colon adenomas and carcinomas. Cancer Res., 47, 2704–2713 (1987).
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