Overexpression of α(1,3)-fucosyltransferase VII is sufficient for the acquisition of lung colonization phenotype in human lung adenocarcinoma HAL-24Luc cells

Abstract

Fuc-TVII transfectants exclusively overexpress sialyl-Le x while Fuc-TIV-transfected cells only overexpress the Le x oligosaccharide. We show that solely Fuc-TVII transfectants are able to adhere to interleukin-1β-stimulated HUVEC monolayers. We also demonstrate that Fuc-TVII overexpression in HAL-24Luc cells is sufficient for the acquisition of the lung colonization phenotype. This is the first report directly showing the contribution of an α(1,3)-Fuc-T to the metastatic behaviour of human lung adenocarcinoma cells. Keywords: metastasis; sialyl-Lewisx ; E-selectin; transfection; human lung adenocarcinoma 1169British Journal of Cancer (1999) 80(8), 1169-1174 © 1999 Cancer Research Campaign Article no. bjoc.1999 Received 6 October 1998 Revised 5 January 1999 Accepted 6 January 1999Correspondence to: J Blanco 1991). Our previous characterization of these cells indicated that the lungs are the only organs colonized when HAL-8Luc cells were inoculated either i.v. or intramuscularly (i.m.) in athymic mice while HAL-24Luc cells did not metastasize. Overexpression of these oligosaccharides correlated with an increased expression of the five α genes and enhanced adhesion to activated endothelial cells. Results from those in vitro adhesion assays pointed to the relevance of the sialyl-Le x oligosaccharide in the binding reaction and indicated a high affinity of this epitope for E-selectin expressed on endothelial cells. These observations reinforced previous findings in which sialyl-Le x had been identified as predicting factor of recurrence in lung adenocarcinoma patients (Ogawa et al, 1994). The aim of the present work was to analyse if α(1,3)-Fuc-TIV and α(1,3)-Fuc-TVII overexpression modifies the adhesive properties and lung colonization potential of non-metastatic human lung adenocarcinoma HAL-24Luc cells.  Cell linesHuman lung adenocarcinoma cell lines HAL-8Luc (metastatic) and HAL-24Luc (non-metastatic) (Martín-Satué et al, 1998) were maintained in RPMI-1640 (Bio-Whitaker, Verviers, Belgium) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS) (Bio-Whitaker), 2 mM L-glutamine (Bio-Whitaker), penicillin (100 units ml -1 ), streptomycin (100 µg ml -1) and 300 µg ml -1 geneticin (G418; Life Technologies, Inc., Grand Island, NY, USA) in a humidified atmosphere of 5% carbon dioxide at 37°C. Human umbilical vein endothelial cells (HUVECs) were isolated by collagenase digestion from multiple segments of normal-term umbilical cords and cultured as previously described (López et al, 1993). Adhesion assays were performed using endothelial cells within three passages. Plasmids and transfectionPlasmids containing full-length cDNAs for Fuc-TIV (pcDNAI-α(1,3)FT Mlu ) and Fuc-TVII (pCDM8-FucTVII) were kindly donated by Dr John B Lowe from the Howard Hughes Medical Institute (The University of Michigan Medical Centre, Ann Arbor, MI, USA). Plasmid pCEP4 encoding hygromycin resistance as well as the pcDNAI and pCDM8 plasmids were obtained from Invitrogen (San Diego, CA, USA).HAL-2...

Transfection of the c‐erbB2/neugene upregulates the expression of sialyl Lewis X, α1,3‐fucosyltransferase VII, and metastatic potential in a human hepatocarcinoma cell line

“…Furthermore, transfection of a1,3FucT-VII cDNA resulted in upregulation of the above metastasis-related phenotypes, and increased cell adhesion to HUVECs and cell migration and invasion in a1,3FucT-VII cDNA-transfected cells were also markedly suppressed by treatment with KM93, but only slightly by FH6 (Table 1). These results further substantiate the close relationship between the expression of a1,3FucT-VII and the metastasis-related phenotypes, and are in accordance with recent reports that SLe x and a1,3FucT-VII play an important role in metastasis of mouse melaloma B16 [41] and human lung adenocarcinoma HAL-24Luc [42]. Furthermore, both the metastasis-associated molecules, SLe x and a1,3FucT-VII, and the metastasis-associated phenotypes, cell adhesion to HUVECs/P-selectin, migration and invasion, correlated positively with the expression of c-erbB2/neu.…”

Carbohydrate‐mediated cell adhesion in cancer metastasis and angiogenesis

“…Fucosyltransferase VII is expressed on epithelial cancer cells, and its significant contribution to sialyl Lewis x synthesis is suggested by several reports. 9,31) A significant increase in FUT7 gene expression has been reported to occur in lung cancers, and this was correlated to sialyl Lewis x expression and the prognosis of patients. 9) However, cells of epithelial origin usually contain a significant amount of transcripts for other fucosyltransferase isoenzymes that are capable of synthesizing sialyl Lewis a/x ; i.e., mainly fucosyltransferases III and VI.…”

Alpha 1,3 fucosyltransferases are master regulators of prostate cancer cell trafficking

“…To this end, we found that ␣1,3 FT elevation was alone sufficient to complete sLe X synthesis and convert a non-E-selectin-binding PC-3 cell to a cell that binds E-selectin. Others have shown that ␣1,3 FTs are associated with progression of several solid cancers of the colon (34)(35)(36), breast (37), lung (38,39), liver (40), and pancreas (41,42). The question is whether PCa cells expressing abnormally high levels of ␣1,3 FTs enter BM at specific microvascular domains where E-selectin and SDF-1␣ are coexpressed constitutively and regulate leukocytic infiltration (9).…”