Задачей следующих экспериментов является продолжение набора данных по экспрессии декорина, а также другого представителя ДСПГ - бигликана, и D-глюкуронил С5-эпимеразы в контрольной и опухолевой ткани молочной железы человека методом вестерн-блота. Также планируется провести определение содержания и локализации данных протеогликанов и D-глюкуронил С5-эпимеразы в тех же образцах ткани методом иммуногистохимии.
ВЫВОДЫ:
1. В опухолевой ткани молочной железы человека происходит увеличение содержания суммарных протеогликанов по сравнению с контрольной тканью в среднем в 4.3 раза, при пересчете на массу ткани, или в среднем в 2 раза при пересчете на количество нуклеиновых кислот в ткани.
2. Увеличение содержания протеогликанов в опухолевой ткани молочной железы человека происходит за счет появления хондроитинсульфат протеогликанов А и С, отсутствующих в контрольной ткани.
3. В опухолевой ткани молочной железы человека изменена структура ГАГ-цепей дерматансульфат протеогликанов. Данная фракция чувствительна к ферменту хондроитиназе АС. Также в опухолевой ткани изменена электрофоретическая подвижность дерматансульфат протеогликана декорина по сравнению с контрольной тканью.
4. В опухолевой ткани молочной железы человека происходит нарушение структуры белковой молекулы D-глюкуронил С5-эпимеразы или полное исчезновение экспрессии данного фермента.
1. Selleck S. B. Overgrowth syndromes and the regulation of signaling complexes by proteoglycans // Am. J. Hum. Genet. 1999. V 64. P.372-377.
2. Rodén L., Koerner T., Olson C. and Schwartz N. B. Mechanisms of chain initiation in the biosynthesis of connective tissue polysaccharides // Fed. Proc. 1985. V.44. P.373-380.
3. Fraser J. R. E., Laurent T. C. and Laurent U. B. G. Hyaluronan: Its nature, distribution, functions and turnover // J. Intern. Med. 1997. V.242. P.27-33.
4. Fraser J. R. and Laurent T. C. Turnover and metabolism of hyaluronan // Ciba Found. Symp. 1989. V.143. P.41-53.
5. DeAngelis P. L. and Achyuthan A. M. Yeast-derived recombinant DG42 protein of Xenopus can synthesize hyaluronan in vitro // J. Biol. Chem. 1996. V.271. P.23657-23660.
6. DeAngelis P. L., Papaconstantinou J. and Weigel P. H. Molecular cloning, identification, and sequence of the hyaluronan synthase gene from group A Streptococcus pyrogenes // J. Biol. Chem. 1993. V.268. P. 19181-19184.
7. Kazuyuki Sugahara, Tadahisa Mikami, Toru Uyama, Souhei Mizuguchiz, Kazuya Nomura and Hiroshi Kitagawa. Recent advances in the structural biology of chondroitin sulfate and dermatan sulfate // Current Opinion in Structural Biology. 2003. V.13. P.612-620.
8. Trowbridge J. M. and Gallo R. L. Dermatan sulfate: new functions from an old glycosaminoglycan // Glycobiology. 2002. V.12. No.9. P.117R-125R.
9. Conrad H. E. Heparin-binding proteins. Academic Press, San Diego. 1998.
10. Salmivirta M., Lidholt K. and Lindahl U. Heparan sulfate: A piece of information // FASEB J. 1996. V.10. P.1270-1279.
11. Tai G. H., Huckerby T. N. and Nieduszynski I. A. Multiple non-reducing chain termini isolated from bovine corneal keratan sulfates // J. Biol. Chem. 1996. V.271. P.23535-23546.
12. Fukuta M., Inazawa J., Torii T., Tsuzuki K., Shimada E. and Habuchi O. Molecular cloning and characterization of human keratan sulfate Gal-6-sulfotransferase // J. Biol. Chem. 1997. V.272. P.32321-32328.
13. Degroote S., Lo-Guidice J. M., Strecker G., Ducourouble M. P., Roussel P. and Lamblin G. Characterization of an N-acetylglucosamine-6-O-sulfotransferase from human respiratory mucosa active on mucin carbohydrate chains // J. Biol. Chem. 1997. V.272. P.29493-29501.
14. Brown G. M., Huckerby T. N., Morris H. G., Abram B. L. and Nieduszynski I. A. Oligosaccharides derived from bovine articular cartilage keratan sulfates after keratanase II digestion: Implications for keratan sulfate structural fingerprinting // Biochemistry 1994. V.33. P.4836-4846.
15. Wilson I. B. The never-ending story of peptide O-xylosyltransferase // Cell. Mol. Life Sci. 2004. V.61. P.794-809.
16. Prydz K., Dalen K. T. Synthesis and sorting of proteoglycans // Journal of Cell Science 2000. V.113. P. 193-205.
17. Silbert J. E., Sugumaran G. Biosynthesis of chondroitin/dermatan sulfate // IUBMB Life. Oct. 2002. V.54 (4). P.177-186.
18. Silbert J. E. and Sugumaran G. Intracellular membranes in the synthesis, transport, and metabolism of proteoglycans // Biochim. Biophys. Acta. 1995. V.1241. P.371-384.
19. Silbert J. E.organization of glycosaminoglycan sulfation in the biosynthesis of proteochondroitin sulfate and proteodermatan sulfate // Glycoconjugate J. 1996. V.13. P.907-912.
20. Fransson L. A., Belting M., Jonsson M., et al. Biosynthesis of decorin and glypican // Matrix Biol. 2000. V. 19. P.367-376.
21. Maccarana M., Sakura Y., Tawada A., Yoshida K. and Lindahl U. Domain structure of heparan sulfates from bovine organs // J. Biol. Chem. 1996. V.271. P.17804-17810.
22. Rosenberg R. D., Shworak N. W., Liu J., Schwartz J. J. and Zhang L. J. Heparan sulfate proteoglycans of the cardiovascular system. Specific structures emerge but how is synthesis regulated // J. Clin. Invest. 1997. V.99. P. 2062-2070.
23. Yanagishita M. and Hascall V. Cell surface heparan sulfate proteoglycans // J. Biol. Chem. 1992. V.267. P.9451-9454.
24. Lyon M., Deakin J. A. and Gallagher J. T. Liver heparan sulfate structure. A novel molecular design // J. Biol. Chem. 1994. V.269. P.11208-11215.
25. Iozzo R. V. Matrix proteoglycans: from molecular design to cellular function // Annu. Rev. Biochem. 1998. V.67. P.609-52.
26. Jackson R. L., Busch S. J. and Cardin A. D. Glycosaminoglycans: Molecular properties, protein interactions, and role in physiological processes // Physiol. Rev. 1991. V.71. P.481-539.
27. Hardingham T. E., Fosang A. J. Proteoglycans: many forms and many functions // The FASEB Journal. 1992. V.6. P.861-870.
28. Tumova S., Woods A., Couchman J. R. Heparan sulfate proteoglycans on cell surface: versatile coordinators of cellular functions // Int J Biochem Cell Biol. 2000. V.32. P.269-288.
29. Selleck S. B. Proteoglycans and pattern formation // TIG. May 2000. V.16. № 5.
30. Perrimor N., Bernfield M. Specificities of heparan sulfate proteoglycans in developmental processes // Nature.13 April2000. V.404.
31. Зимина Н.П., Рыкова В.И. Протеогликаны животных тканей и их влияние на синтез ДНК // Биохимия. 1986. Т.51. Вып.9. С.1555-1561.
32. Рыкова В.И., Григорьева Э.В. Состав протеогликанов в ядрах клеток гепатомы мышей // Биохимия. 1998. Т.63. Вып.11. С.1496-1502.
33. Schamhart D. H., Kurth K. H. Role of proteoglycans in cell adhesion of prostate cancer cells: from review to experiment // Urol Res. 1997. V.25. P.89-96.
34. Schonherr E., Witsch-Prehm P., Harrach B., et al. Interaction of biglycan with type I collagen // J Biol Chem. 1995. V.270. P.2776-2783.
35. Bianco P., Fisher L. W., Young M. F., et al. Expression and localization of the two small proteoglycans biglycan and decorin in developing human skeletal and non-skeletal tissues // J Histochem Citochem. 1990. V.38. P.1549-1563.
36. Fleischmajer R., Fisher L. W., MacDonald E. D., et al. Decorin interacts with fibrillar collagen of embryonic and adult human skin // J Struct Biol. 1991. V.1. P.82-90.
37. Oldberg A., Antonsson P., Lindblom K., et al. A collagen-binding 59-kd protein (fibromodulin) is structurally related to the small interstitial proteoglycans PG-S1 and PG-S2 (decorin) // EMBO J. 1989. V.8. P.2601-2604.
38. Embry J. J., Knudson W. G1 domain of aggrecan cointernalizes with hyaluronan via a CD44-mediated mechanism in bovine articular chondrocytes // Arthritis Rheum. 2003. V.12. P.3431-3441.
39. Matsumoto K., Shionyu M., Go M., et al. Distinct interaction of versican/PG-M with hyaluronan and link protein // J Biol Chem. 2003. V.42. P.41205-41212.
40. Hirose J., Kawashima H., Yoshie O., et al. Versican interacts with chemokines and modulates cellular response // J Biol Chem. 2001. V.276. P.5228-5234.
41. Kawashima H., Li Y. F., Watanabe N., Hirose J., et al. Identification and characterization of ligands for L-selectin in the kidney.I. Versican, a large chondroitin sulfate proteoglycan, is a ligand for L - selectin // Int Immunol. 1999. V.3. P.393-405.
42. Kawashima H., Hirose M., Hirose J., et al. Binding of large chondroitin sulfate/dermatan sulfate proteoglycan, versican, to L-selectin, P-selectin and CD 44 // J Biol Chem. 2000. V.275. P.35448-35456.
43. Haggerty J. G., Bretton R. H., Milstone L. M. Identification and characterization of cell surface proteoglycan on keratinocytes // J Invest Dermatol. 1992. V.4. P.374-380.
44. Kugelman L. C., Ganguly S., Haggerty J. G., et al. The core protein of epican, a heparan sulfate proteoglycan on keratinocytes, is an alternative form of CD 44 // J Invest Dermatol. 1992. V.6. P.866-891.
45. Brown T. A., Bouchard T., St John T., et al. Human keratinocytes express a new CD 44 core protein (CD 44) as a heparin-sulfate intrinsic membrane proteoglycan with additional exon // J Cell Biol. 1991. V.1. P. 207-221.
46. Kolset S. O. and Gallagher J. T. Proteoglycans in haemopoietic cells // Biochim. Biophys. Acta. 1990. V.1032. P. 191-211.
47. Santra M., Reed C. C., Iozzo R. V. Decorin binds to a narrow region of the epidermal growth factor (EGF) receptor, partially overlapping but distinct from the EGF-binding epitope // J Biol Chem. 2002. V.38. P.35671-35681.
48. Eickelberg O., Cerntrella M., Reiss M., et al. Betaglycan inhibits TGF-beta signaling by preventing type I-type II receptor complex formation. Glycosaminoglycan modifications alter betaglycan function // J Biol Chem. 2002. V.277. P.823-829.
49. Ostrovsky O., Berman B., Gallagher J., et al. Differential effects of heparin saccharides on the formation of specific fibroblast growth factor (FGF) and FGF receptor complexes // J Biol Chem. 2002. V.4. P.2444-2453.
50. Wu Z. L., Zhang L., Yabe T., et al. The involvement of HS in FGF1/HS/FGFR1 signaling complex // J Biol Chem. 2003. V. 19. P.17121-17129.
51. Rouslahti E. Proteoglycans in cell regulation // J Biol Chem. 1989. Aug 15. V.264 (23). P.13369-72.
52. Iozzo R. V.,Cohen I.Altered proteoglycan gene expression and the tumor stroma // Experientia. 1993. V.49. P.447-455.
53. Lesley J., Hyman R., English N., Catterall J. B., Turner G. A. CD44 in inflammation and metastasis // Glycoconj. J. 1997. V.14. P.611-622.
54. JozzoR. V. Neoplastic modulation of extracellular matrix. Colon carcinoma cells release polypeptides that alter proteoglycan metabolism in colon fibroblasts // J Biol Chem. 1985 Jun 25. V.260 (12). P.7464-73.
55. Theocharis A. D., Vynios D. H., Papageorgacopoulou N., et al. Altered content, composition and structure of glycosaminoglycans and proteoglycans in gastric carcinoma // Int J Biochem Cell Biol. 2003. V.35 (3). P.376-390.
56. Theocharis A. D., Tsara M. E., Papageorgacopoulou N., et al. Pancreatic carcinoma is characterized by elevated content of hyaluronan fnd chondroitin sulfate with altered disaccharide composition // Biochem Biophys Acta. 2000. V.2. P. 201-206.
57. Tsara M. E., Theocharis A. D., Theocharis D. A.compositional and structural alterations of proteoglycans in human rectum carcinoma with special reference to versican and decorin // Anticancer Res. 2002. V.22. P.2893-2898.
58. Achilleas D, Theocharis. Human colon adenocarcinoma is associated with specific post-translational modifications of versican and decorin // Biochem Biophys Acta. 2002. V.1588. P.165-172.
59. Papadas T. A., Stylianou M., Mastronikolis N. S., et al. Alterations in the content and composition of glycosaminoglycans in human laryngeal carcinoma // Acta Otolaryngol. 2002. V.122. P.330-337.
60. Vijayagopal P., Figueroa J. E., Levine E. A., et al. Altered composition and increased endothelial cell proliferative activity of proteoglycans isolated from breast carcinoma // J Surg Oncol. 1998. V.4. P.250-254.
61. Alessandra G. A., Berto A., Lucia O., et al. A comparative analysis of structure and spatial distribution of decorin in human leiomyoma and normal myometrium // Biochem Biophys Acta. 2003. V.1619. P.98-112.
62. Ladanyi A., Gallai M., Paku S., Nagu J., O.,DudasJ.,TimarJ.,KovalszkyI. Expression of a decorin-like moleculein human melanoma // Pathol Oncol Res. 2001. V.7 (4). P.260-6.
63. SantraM., Reed C. C., Iozzo R. V. Decorin binds to a narrow region of the epidermal growth factor (EGF) receptor, partially overlapping but distinct from the EGF-binding epitope // J. Biol. Chem. 2002. V.38. P.35671-3581.
64. De Luca A. Santra, M. Baldi A., et al. Decorin-induced growth suppression is associated with up-regulation of p21, an inhibitor of cyclin-dependent kinases // J. Biol. Chem. 1996. V.271. P.18961-18965.
65. Csordas G., Santra M., Reed C. C. et al. Sustained down-regulation of the epidermal growth factor receptor by decorin. A mechanism for controlling tumor growth in vivo // J. Biol. Chem. 2000. V.42. P.32879-32887.
66. Stander M., Naumann U.,Wick W., et al. Transforming growth factor-beta and p-21: multiple molecular targets of decorin-mediated suppression of neoplastic growth // Cell Tissue Res. 1999. V.296. P.221-227.
67. Clouthier D. E., Comerford S. A., Hammer R. E. Hepatic fibrosis, glomerulosclerosis, and a lipodystrophy-like syndrome in PEPK-TGF-beta 1 transgenic mice // J. Clin. Invest. 1998. V.100. P.2697-26713.
68. SantraM., Eichstetter I., Iozzo R. V. An anti-oncogenic role for decorin. Down-regulation of ErbB2 leads to growth suppression and cytodifferentiation of mammary carcinoma cells // J. Biol. Chem. 2000. V.45. P.35153-35161.
69. Grant D. S., Yenisey C., Rose R. W. et al. Decorin suppresses tumorcell-mediated angiogenesis // Oncogene. 2002. V.31. P.4765-4777.
70. Schonherr E., Levkau B., Schaefer L. et al. Decorin affects endothelial cells by Akt-dependent and - independent pathways // Ann. N. Y. Acad. Sci. 2002. V.973. P.149-152.
71. NashM. A., Loercher A. E., FreedmanR. S. In vitro growth inhibition of ovarian cancer cells by decorin: synergism of action between decorin and carboplatin // Cancer Res. 1999. V.59. P.6192-6199.
72. Santra, M., Skorski, T., Calabretta, B., et al. De novo decorin gene expression suppresses the malignant phenotype in human colon cancer cells // Proc. Natl. Acad. Sci. - 1995. - V.92. - P.7016-7020.
73. ReedC. C., Gauldie J., Iozzo R. V. Suppression of tumorigenicity by adenovirus-mediated gene transfer of decorin // Oncogene. - 2002. V.23. P.3688-95.
74. Tiedemann K., Larsson T.,Heinegard D., et al. The glucuronyl C5-epimerase activity is the limiting factor in the dermatan sulfate biosynthesis // Arch. Biochem. Biophys. 2001. V.391. P.65-71.
75. Hagner-McWhirterA.,Li J. Irreversible Glucuronyl C5-epimerization in the Biosynthesis of Heparan Sulfate // J. Biol. Chem. 2004. V.279. P.14631-14638.
76. Hagner-McWhirter A.,Li J. P.,Oscarson S., et al. Irreversible glucuronyl C5-epimerization in the biosynthesis of heparan sulfate // J. Biol. Chem. 2004. V.279. P.14631-14638.