[Transfection of alpha1, 2-fucosyltransferase gene increases the antigenic expression of Lewis y in ovarian cancer cell line RMG-I].
OBJECTIVE
To transfect human alpha1, 2-fucosyltransferase (alpha1, 2-FT) gene to ovarian cancer cell line RMG-I and investigate the antigenic expression change of Lewis y and the other related oligosaccharides.


METHODS
The expression vector pcDNA3.1(-)-HFUT-H was constructed by polymerase chain reaction (PCR) to clone human alpha1, 2-FT gene coding region. The alpha1, 2-FT gene stable high-expression cell line RMG-I-H was established by transfecting pcDNA3.1(-)-HFUT-H to ovarian cell line RMG-I. The change of alpha1, 2-FT activity in the cell line before and after the tranfection was confirmed by the determination of enzymatic activity. The changes of cell lipid and glucolipid, especially the change of type II oligosaccharide, in the cell line before and after the transfection was determined by Thin-Layer Chromatography (TLC) and TLC immunostaining method, respectively.


RESULTS
The H-1 antigen and Lewis y antigen were obviously increased in the cell line RMG-1-H, especially the latter one, which was 20 times higher than before, and the type I saccharide chain Lewis b was decreased significantly. The main lipid components on the cell membrane, cholesterol and phosphatides, showed no change in the cell lines before and after the transfection, and the neutral glycolipid also showed no obvious change.


CONCLUSIONS
The transfection of alpha1, 2-FT gene can increase the activity of alpha1, 2-FT in the cell line RMG-I and mainly increase the expression of Lewis y antigen simultaneously. The construction of RMG-I Lewis y high expression cell line provides a cell model for further study on the relationship between Lewis y antigen and biological behaviors in the ovarian cancer.
Lin
Bei
B
Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, Shenyang 110004, China. linbei88@hotmail.com
Hao
Ying-Ying
YY
Wang
Dong-Dong
DD
Zhu
Lian-Cheng
LC
Zhang
Shu-Lan
SL
Saito
Makiko
M
Iwamori
Masao
M
D045744:Cell Line, Tumor; D002855:Chromatography, Thin Layer; D005260:Female; D005647:Fucosyltransferases / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007983:Lewis Blood Group Antigens / Q000378:metabolism*; D010051:Ovarian Neoplasms / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D007983:Lewis Blood Group Antigens; C094162:Lewis Y antigen; D005647:Fucosyltransferases