Aberrant glycosylation of cell surface glycoproteins is commonly associated with the invasion and metastasis of colorectal carcinomas, which can be attributed to the upregulated expression of glycosyltransferases. Therefore, elucidation of glycosyltransferases and their substrates may improve our understanding of their roles in tumor metastasis. β-1,3-N-acetylglucosaminyltransferase-8 (β3GnT8) is a key enzyme that catalyzes the formation of poly-N-acetyllactosamine (polylactosamine) chains on β1,6-branched N-glycans in vitro, which is also involved in tumor invasion. In the present study, we analyzed the expression of β3GnT8 and its product polylactosamine in four human colorectal carcinoma cell lines (LS-174T, SW620, SW480 and LoVo) with different metastatic potential. We found that the levels of β3GnT8 and polylactosamine chains were gradually increased in the colorectal cancer cell lines in a trend from low to high metastatic potential. Notably, a significantly positive relationship between β3GnT8 expression and HG-CD147 was noted in the colorectal cancer cell lines. To further investigate their relationships, exogenous β3GnT8 was introduced into the LS-174T cells, while expression of β3GnT8 was downregulated in the LoVo cells. The overexpression of β3GnT8 in LS-174T cells increased the level of HG-CD147. Conversely, downregulation of β3GnT8 expression in LoVo cells significantly decreased the expression of HG-CD147. HG-CD147 is a major carrier of β1,6-branched polylactosamine sugars; therefore, the regulation of β3GnT8 significantly altered the β1,6-branched polylactosamine structures on CD147. Hence, we suggest that β3GnT8 plays a key role in the metastasis of colorectal cancer cells by altering the β1,6-branched polylactosamine sugars of CD147.

Abstract. 5-Fluorouracil (5-FU) is the major chemotherapeutic agent for the treatment of colorectal carcinoma, which were found to have N-glycans containing polylactosamine on the cancer cell surface. Alterations in the expression and structure of polylactosamine glycans are associated with cellular differentiation and oncogenesis. However, little is known with regard to the correlation between the levels of polylactosamine expressed in colon cancer cells and the anticancer effect of 5-FU. In the present study, SW620 cells were treated with the half maximal inhibitory concentration (IC 50 ; determined by MTT-assay) of 5-FU. Hoechst 33258 staining and flow cytometric analysis indicated that 5-FU administration resulted in apoptosis in SW620 cells. An increased percentage of cells in S phase was also observed among the SW620 cells treated with 5-FU. Under the same experimental conditions, a decrease in the 5-FU-induced inhibition of polylactosamine glycans was recorded. However, an increase in the activity of alkaline phosphatase was also observed. Furthermore, pretreatment of the SW620 cells with 5-FU inhibited the expression of β1,3-N-acetylglucosaminyltran sferase-8 (β3Gn-T8) and cluster of differentiation (CD)147 in a time-dependent manner. Overall, changes in glycosylation were associated with the anticancer effect of 5-FU in the colon cancer cells. In conclusion, polylactosamine may be a useful target for the identification of substances with anticancer activity.

Abstract. aber rant glycosylation is known to be associated with cancer chemoresistance. β-1,3-n-acetylglucosaminyltransferase (β3gnt)8, which synthesizes polylactosamine on β1-6 branched n-glycans, is dramatically upregulated in colorectal cancer (crc). 5-fluorouracil (5-fu) resistance remains a major obstacle to the chemotherapy of crc. However, little is known with regard to the correlation between 5-fu resistance and the expression of β3gnt8 in crc. in this study, a 5-fu-resistant cell line (SW620/5-fu) was generated, and 50% inhibition concentration (ic50) of 5-fu was determined by Mtt assay. flow cytometry and lectin blot analysis were performed to detect the alteration of polylactosamine structures. Quantitative rt-Pcr and western blot analysis were used to identify and evaluate candidate genes involved in the synthesis of polylactosamine in SW620/5-fu cells. We found polylactosamine chains were significantly increased in SW620/5-fu cells. inhibition of the biosynthesis of polylactosamine by 3'-azidothymidine (aZt) was able to reduce 5-fu tolerance. further studies showed that β3gnt8 expression was also upregulated in 5-fu-resistant cancer cells, and knockdown of β3gnt8 by rna interference reversed 5-fu resistance through, at least partly, by suppressing the formation of polylactosamine. in conclusion, the alteration of β3gnt8 in crc cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies.

β1, 3-N-acetylglucosminyltransferase 8(β3GnT8) synthesizes a unique cabohydrate structure known as polylactosamine, and plays a vital role in progression of various human cancer types. However, its involvement in gastric cancer remains unclear. In this study, we analyzed the expression and clinical significance of β3GnT8 by Western blot in 6 paired fresh gastric cancer tissues, noncancerous tissues and immunohistochemistry on 110 paraffin-embedded slices. β3GnT8 was found to be over-expressed in gastric cancer tissues, which correlated with lymph node metastasis and TNM stage. Forced the expression of β3GnT8 promoted migration and invasion of gastric cancer cells, whereas β3GnT8 knockdown led to the opposite results. Further studies showed that the regulated β3GnT8 could convert the heterogeneous N-glycosylated forms of CD147 and change the polylactosamine structures carried on CD147. In addition, our data suggested the annexin A2 (ANXA2) to be an essential interaction partner of β3GnT8 during the process of CD147 glycosylation. Collectively, these results provide a novel molecular mechanism for β3GnT8 in promotion of gastric cancer invasion and metastasis. Targeting β3GnT8 could serve as a new strategy for future gastric cancer therapy.

Active magnetic bearing system is a typical geometric constraint system which is combined with each part by certain assembly constraint relations. Graph theory is used to build the assembly model in the progress of assembly design. Trough the graph to express this system, it is not only clear and intuitive, and the algorithm is stable and high efficiency. Modeling active magnetic bearing assembly system is achieved by hierarchical model of data structure, which leads to accelerate the assessment of system configuration program and save a lot of time for designers.