Characteristic Expression of Lewis-antigenic Glycolipids in Human Ovarian Carcinoma-derived Cells with Anticancer Drug-resistance

Abstract

By comparing ovarian carcinoma-derived KF28 cells with the corresponding anticancer drug-resistant cells, the taxol- and cisplatin-resistant properties were found to be closely related with MDR1 and BSEP, and MRP2 transporters, respectively. In addition to the transporters expression, the amounts of glycolipids, particularly their longer carbohydrate structures, in the resistant cells increased to 3-4-fold of those in the sensitive cells due to enhanced transcription of the respective glycosyltransferases. The major glycolipids in the sensitive and resistant cells were GlcCer and Gb(3)Cer, respectively, and extension of the carbohydrate structure into Lewis antigen characteristically occurred in the resistant cells. Le(b), which was not detected in the cisplatin-resistant cells, was present in the taxol-resistant cells, while Le(x) was present in the cisplatin-resistant cells at a higher concentration than in the taxol-resistant cells. 2-Hydroxy fatty acids were significantly abundant in glycolipids of the resistant cells, but they were not detected in free ceramides or sphingomyelin, indicating that the enhanced synthesis of glycolipids in the resistant cells was not linked with the removal pathway for virulent ceramides derived from sphingomyelin. The resistant cells with abundant glycolipids exhibited lower membrane fluidity than the KF28 cells, and this property might be involved in the anticancer drug-resistance.

Enhanced expression of unique gangliosides with GM2-determinant in human uterine cervical carcinoma-derived cell lines

“…We have also observed that Le X and Le b are characteristically expressed in ovarian serous carcinoma KF28 cells exhibiting anti-cancer drug resistance, and enhanced expression of Le Y in ovarian clear cell carcinoma RMG-1 cells on transfection of α1,2-fucosyltransferase results in high dissemination potential and anti-cancer drug resistance [10,11]. On comparison of glycolipid compositions between KF28 cells and KF28-derived anticancer drug-resistant cells, and between RMG-1 cells and fucosyltransferase gene-transfected RMG-1 cells, globo-, lacto-and neolacto-series glycolipid profiles were found to be characteristically changed to cause alteration of the antigenicity of glycolipids, and their cellular functions including in anticancer drug-resistance and peritoneal dissemination [11,12]. In addition, on comparison of glycolipids in tissues and cells of histologically defined ovarian carcinomas, i.e., serous, mucinous, endometrioid and clear cells, the amounts of sulfatides in the mucinous type were found to be strikingly higher than those in the other types, but the expression of globo-and ganglio-series glycolipids including Lewis antigens was not related with the above histological classifications [13][14][15].…”

“…4a and b. Thus, enhanced expression of gangliosides with GM2 determinant, particularly with unique structures, GalNAc-GM1b and GalNAc-GD1a, was a characteristic of cervical carcinomaderived cell lines, which was not observed for endometrial and ovarian carcinoma-derived cell lines [11,15].…”

“…Analysis of glycolipids Lipids were extracted from lyophilized tissues and cells with chloroform/methanol/water [10][11][12][13][14]. For structural analysis, individual gangliosides were purified from the acidic lipids using a silica gel (Iatrobeads 6RS8060: Iatron Laboratory, Tokyo) column by gradient elution with chloroform/methanol/water (55:45: 2 and 10:90:4, by vol.).…”

“…sialyl Le a antigen for diagnosis of pancreatic, gallbladder and colon carcinomas [5,6], and Le b , Le Y and H antigens for prediction of metastatic potential [7][8][9]. We have also observed that Le X and Le b are characteristically expressed in ovarian serous carcinoma KF28 cells exhibiting anti-cancer drug resistance, and enhanced expression of Le Y in ovarian clear cell carcinoma RMG-1 cells on transfection of α1,2-fucosyltransferase results in high dissemination potential and anti-cancer drug resistance [10,11]. On comparison of glycolipid compositions between KF28 cells and KF28-derived anticancer drug-resistant cells, and between RMG-1 cells and fucosyltransferase gene-transfected RMG-1 cells, globo-, lacto-and neolacto-series glycolipid profiles were found to be characteristically changed to cause alteration of the antigenicity of glycolipids, and their cellular functions including in anticancer drug-resistance and peritoneal dissemination [11,12].…”

Expression of α2,6-sialic acid-containing and Lewis-active glycolipids in several types of human ovarian carcinomas

“…The neutral and acidic glycolipids derived from the tissues and cells were examined by TLC and TLC-immunostaining with the development solvents, chloroform/methanol/water (65:35:8, by volume) for neutral glycolipids and chloroform/methanol/0.5% CaCl 2 (55:45:10, by volume) for gangliosides, as previously described (10,13). Known amounts (0.1-1.5 µg) of glycolipids, such as GM3, Lc 4 Cer, IV 3 NeuAcα-nLc 4 Cer, IV 6 NeuAcα-nLc 4 Cer and GD3, were developed on the same TLC plates for the preparation of standard curves.…”