Changes in the invasive and metastatic capacities of HT-29/M3 cells induced by the expression of fucosyltransferase 1

Abstract

Lewis antigens are terminal fucosylated oligosaccharides synthesized by the sequential action of several glycosyltransferases. The fucosyltransferases are the enzymes responsible for the addition of terminal fucose to precursor oligosaccharides attached to proteins or lipids. These oligosaccharides, defined as cell surface markers, have been implicated in different types of intercellular interactions and in adhesion and invasion processes. Transfection of HT-29/M3 colon cancer cells with the full length of human fucosyltransferase (FUT1), induces the synthesis of H type 2 and Lewis y antigens, associated with a decrease of sialyl-Lewis x. The capacity to develop primary tumors when cells were injected intrasplenically was similar in parental and FUT1-transfected cells, but the capacity to colonize the liver after spleen removal was significantly reduced in M3/FUT1 transfected cells. These results indicate that the expression of FUT1 induces changes in the metastatic capacity of HT-29/M3 colon cancer cells, as a consequence of the altered expression pattern of type 2 Lewis antigens. Also, an association between MUC5AC expression and the degree of gland differentiation in both primary splenic tumors and hepatic metastases was detected.

Figure 1

(a) Expression of H type 2, Sialyl‐Lewis a (s‐Le a), and s‐Le x antigens in M3‐4b parental cells and C50‐6a/fucosyltransferase (FUT1)‐transfected cells detected by flow cytometry. (b) Treatment of C50‐6a/FUT1‐transfected cells with phenyl β‐D‐galactoside: Flow cytometry detection of H type 2 antigen, s‐Le a, and s‐Le x after 4 days of treatment. Marker 1 (M1) indicates the positive cells.

Figure 2

Binding of M3‐4b parental cells and C50‐6a/ fucosyltransferase (FUT1)‐transfected cells to E‐Selectin. The two cell lines were also incubated with anti‐s‐Le x (KM93) and anti‐s‐Le a (57/27) antibodies. **P = 0.05.

Figure 3

MUC5AC, and Le y expression in primary splenic tumors after the injection of M3‐4b parental cells and C50‐6a/FUT1‐transfected cells, detected by immunohistochemistry. Original magnification × 200.

Figure 4

H type 2 antigen, Sialyl‐Lewis a (s‐Le a), and s‐Le x expression in liver metastases after the intrasplenic injection of HT‐29/M3 and C50‐6a/FUT1 cells, detected by immunohistochemistry. Original magnification × 200.