High‐performance capillary electrophoretic analysis of hyaluronan and galactosaminoglycan‐disaccharides in gastrointestinal carcinomas. Differential disaccharide composition as a possible tool‐indicator for malignancies

Theocharis, Achilleas D.; Theocharis, Dimitrios A.

doi:10.1002/bmc.145

Cited by 32 publications

(19 citation statements)

References 18 publications

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“…In addition, the molecular mass of the chains (Table III) showed a stage-related decrease, indicating the presence of increasing levels of core protein substrates of the chondroitinsynthesizing enzymes. These observations are in agreement with previous findings (11,15,17,18), and suggest a very high biosynthetic rate of CS/DS in cancer, which may be attributable to the increased expression of the related biosynthetic enzymes.…”

Section: Resultssupporting

confidence: 93%

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Chondroitin synthases I, II, III and chondroitin sulfate glucuronyltransferase expression in colorectal cancer

et al. 2011

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Abstract. Glycosaminoglycans undergo significant structural alterations in cancer, namely in terms of their sulfation pattern and hydrodynamic size. Numerous studies have focused on this issue, and have demonstrated that glycosaminoglycans play a crucial role in cancer growth and invasion. However, the majority of the enzymes involved in glycosaminoglycan alterations have yet to be examined in detail. The present study focused on the expression of chondroitin-synthesizing enzymes in colorectal cancer. Specimens from healthy controls and cancer patients were subjected to RT-PCR analysis after RNA isolation, and to Western blotting after sequential extraction. The results indicated that chondroitin polymerizing factor and glucuronyltransferase gradually increased with cancer stage, and were expressed at much higher levels in adenomas compared to adjacent normal tissue. The opposite profile was obtained for chondroitin synthase I. Chondroitin synthase III was present at low levels in all the samples examined; however, its expression was higher in the samples from the cancer patients than in those from the healthy controls. It can therefore be concluded that, among the various factors regulating the structure of glycosaminoglycans in cancer, the differential expression of chondroitin-synthesizing enzymes is of the most significance.

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Section: Resultssupporting

confidence: 93%

“…In general, glycosaminoglycans are altered in cancer in qualitative and quantitative terms (10)(11)(12)(13)(14)(15)(16)(17)(18). In the majority of malignancies, CS levels are increased.…”

Section: Introductionmentioning

confidence: 99%

Chondroitin synthases I, II, III and chondroitin sulfate glucuronyltransferase expression in colorectal cancer

et al. 2011

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“…Additionally, non-sulfated chondroitin is detected in cancerous tissues, which is almost absent in normal specimens. Furthermore, the total content of chondroitinase ABC resistant GAGs in cancerous tissues is decreased compared to the normal ones (2) and differences in the sulfation pattern of Hep are also described (3). In adenomas, HS contains more sulfate groups than in carcinomas.…”

Section: Introductionmentioning

confidence: 99%

“…Much research has focused on glycosaminoglycans in a variety of tumors of mainly the gastrointestinal tract. In poorly differentiated tumors a great increase of hyaluronan and Δdi-mono6S has been observed (2). Additionally, non-sulfated chondroitin is detected in cancerous tissues, which is almost absent in normal specimens.…”

Section: Introductionmentioning

confidence: 99%

Alterations of glycosaminoglycan disaccharide content and composition in colorectal cancer: Structural and expressional studies

Kalathas

Theocharis²,

Bounias³

et al. 2009

Oncol Rep

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Abstract.The glycosaminoglycans are implicated in many processes important in the growth and progression of malignant tumors. In the present study glycosaminoglycans were purified from healthy, macroscopically normal and cancerous specimens of different anatomic sites and different stages of cancer and analyzed by FACE after chondroitinases and sulfatases digestion. The cancerous samples contained increased levels of 6-sulfated unsaturated disaccharides compared to macroscopically normal and healthy samples, the increase being stage-related. The differences in sulfation were found to be related to the anatomic site and the stage of cancer. RT-PCR analysis of 4-sulfotransferase mRNA revealed its presence in decreasing amounts as the stage of the cancer increased. Furthermore, the percent content of hyaluronan disaccharides was elevated in macroscopically normal samples compared to the cancerous, and in addition, it was much more elevated than that of healthy samples. Haluronan levels increase with stage in cancerous tissues. Therefore, it could be concluded that the glycosaminoglycans in colorectal cancer are biosynthetically directed to contribute in different ways depending on the cancer stage and anatomical site.

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“…1), contain unsaturated uronic acid residues at their nonreducing end allowing sensitive detection at 232 nm [1-3, 19, 26, 27]. This method has also been applied to the determination of HA of gastrointestinal carcinomas [28], and the CE analysis of GAG-derived disaccharides in biological samples has been reviewed by Lamari et al [3]. HA has no unsaturated glucuronic acid and can thus only be detected at 190-200 nm, an approach that provides lower sensitivity and is prone to more interferences when analyzing biological samples.…”

Section: Introductionmentioning

confidence: 99%

Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking

Yang

Breadmore

Thormann³

2005

J of Separation Science

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Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking CE conditions for monitoring the unsaturated disaccharides of hyaluronic acid (di-HA) and chondroitin sulfate (di-CS) using an alkaline tetraborate buffer, electrokinetic sample injection, and UV absorption detection at 232 nm are reported. Separations were performed in an uncoated fused-silica capillary having reversed polarity and reversed electroosmosis generated with the addition of CTAB to the buffer. The influence of various separation parameters, including the concentration of CTAB, buffer pH, concentration of tetraborate, and applied voltage, on the resolution of the two disaccharides was investigated. Baseline separation was obtained with 25 mM tetraborate at pH 10.0 and having 0.05 mM CTAB. Chloride and phosphate in the sample are beneficial for the stacking of the disaccharides, with di-HA forming a much sharper peak than di-CS. Using samples prepared in 25 mM Tris-HCl (pH 7.5) and electrokinetic injection at the cathode at -10 kV for 40 s, linear relationships between the corrected peak area and the concentration of the disaccharides have been found in the ranges of 1.0-400.0 and 0.1-1.0 lg/mL (0.2-1.0 lg/mL for di-CS), with correlation coefficients being A0.9933 in all cases. The RSDs of detection times and corrected peak areas were between 1.13-1.24 and 1.57-2.13%, respectively. Applied to human serum samples that were prepared by ethanol precipitation and depolymerization of the two polysaccharides with chondroitinase ABC reveals comigration of endogenous compounds with di-HA and a sample-dependent detection time. The di-HA content in the serum sample can be estimated via subtraction of the blank peak that is obtained without enzymatic hydrolysis.

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High‐performance capillary electrophoretic analysis of hyaluronan and galactosaminoglycan‐disaccharides in gastrointestinal carcinomas. Differential disaccharide composition as a possible tool‐indicator for malignancies

Cited by 32 publications

References 18 publications

Chondroitin synthases I, II, III and chondroitin sulfate glucuronyltransferase expression in colorectal cancer

Chondroitin synthases I, II, III and chondroitin sulfate glucuronyltransferase expression in colorectal cancer

Alterations of glycosaminoglycan disaccharide content and composition in colorectal cancer: Structural and expressional studies

Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking

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