Ciołczyk-Wierzbicka Dorota, Bodzioch Marek, Gil Dorota, Zmudzińska Danuta, Dembińska-Kieć Aldona, Laidler Piotr
Department of Medical Biochemistry, Medical College, Jagiellonian University, M Kopernika 7, 31-034, Kraków, Poland.
Med Chem. 2007 Sep;3(5):418-24. doi: 10.2174/157340607781745401.
During carcinogenesis aberrant N-glycosylation may lead to the development of subpopulations of tumor cells with altered adhesion properties and increased invasive potential. Biosynthesis of glycans and oligosaccharides is tissue-specific and developmentally regulated by number of glycosyltransferases of which fucosyl-, sialyl- and N-acetylglucosaminyltransferases often participate in synthesis of tumor type glycans. We analyzed the expression of selected glycosyltransferases (real-time PCR): fucosyltransferases FUT-1 and FUT-4, sialyltransferase SIAT4C and beta 1,6-N-acetylglucosaminyltransferase V (MGAT-5), in human melanoma cell lines: WM35 from primary tumor site and WM239, WM9, A375 from metastatic sites. In parallel their proliferation (crystal violet test) and adhesion to fibronectin and collagen IV (BD Biocoat assay) was assessed. Examined cell lines showed expression of all studied glycosyltransferases. The level of expression of fucosyltransferases was significantly higher in melanoma cell lines from metastatic site than from primary cell line: mRNA expression of FUT-1 was 100 times higher in A375 melanoma cell line from metastatic site (A375, solid tumor) than in WM35 primary cell line. The expression of FUT-4 in cell lines from metastatic sites: WM9 (lymph node) and WM239 (skin) was respectively 80 and 37 times higher than in WM 35 primary cell line. In all melanoma cell lines very low expression of MGAT-5 and high expression of SIAT4C was observed. Melanoma cells bound both to fibronectin and to collagen IV. LTA (Lotus tetragonolobus agglutinin), the lectin that specifically recognizes fucose residue of glycans and 20mM L-fucose by itself significantly reduced adhesion of all studied cell lines, both primary and metastatic, to fibronectin (20-50 %) and to collagen IV (20-50 %). In addition LTA reduced the proliferation (20-30 %) of metastatic cell lines (A375, WM9, WM239) and did not affect the growth of primary cell line (WM35). The results suggest that higher expression of fucosyltransferases (FUT-1, FUT-4) might be an important step in the formation of surface structures that facilitate metastasis of melanoma.
在致癌过程中，异常的N-糖基化可能导致具有改变的黏附特性和增加的侵袭潜能的肿瘤细胞亚群的发展。聚糖和寡糖的生物合成具有组织特异性，并受到多种糖基转移酶的发育调控，其中岩藻糖基转移酶、唾液酸转移酶和N-乙酰葡糖胺基转移酶经常参与肿瘤类型聚糖的合成。我们分析了所选糖基转移酶（实时PCR）的表达：岩藻糖基转移酶FUT-1和FUT-4、唾液酸转移酶SIAT4C和β1,6-N-乙酰葡糖胺基转移酶V（MGAT-5），在人黑色素瘤细胞系中：来自原发性肿瘤部位的WM35以及来自转移部位的WM239、WM9、A375。同时评估了它们的增殖（结晶紫试验）以及对纤连蛋白和IV型胶原的黏附（BD生物包被试验）。所检测的细胞系均显示出所有研究的糖基转移酶的表达。转移部位的黑色素瘤细胞系中岩藻糖基转移酶的表达水平显著高于原发性细胞系：来自转移部位（A375，实体瘤）的A375黑色素瘤细胞系中FUT-1的mRNA表达比WM35原发性细胞系高100倍。来自转移部位的细胞系WM9（淋巴结）和WM239（皮肤）中FUT-4的表达分别比WM35原发性细胞系高80倍和37倍。在所有黑色素瘤细胞系中均观察到MGAT-5的低表达和SIAT4C的高表达。黑色素瘤细胞既能与纤连蛋白结合，也能与IV型胶原结合。LTA（四角豆凝集素），一种特异性识别聚糖中岩藻糖残基的凝集素，其本身以及20mM L-岩藻糖显著降低了所有研究的细胞系（原发性和转移性）对纤连蛋白（20%-50%）和IV型胶原（20%-50%）的黏附。此外，LTA降低了转移性细胞系（A375、WM9、WM239）的增殖（20%-30%），而不影响原发性细胞系（WM35）的生长。结果表明，岩藻糖基转移酶（FUT-1、FUT-4）的高表达可能是促进黑色素瘤转移的表面结构形成中的重要一步。
