Expression of Globo H and SSEA3 in breast cancer stem cells and the involvement of fucosyl transferases 1 and 2 in Globo H synthesis.
We examined the expression in breast cancer stem cells (BCSCs) of Globo H, a potential tumor-associated antigen for immunotherapy of epithelial cancers including breast cancer. Flow cytometry revealed Globo H expression in 25/41 breast cancer specimens (61.0%). Non-BCSCs from 25/25 and BCSCs from 8/40 (20%) expressed Globo H. We showed the expression of stage-specific embryonic antigen 3 (SSEA3), the pentasaccharide precursor of Globo H, in 31/40 (77.5%) tumors. Non-BCSCs from 29/40 [corrected] and BCSCs from 25/40 (62.5%) expressed SSEA3. Like Globo H, SSEA3 expression in normal tissues was predominately at the secretory borders of epithelium, where access to the immune system is restricted. Immunization of mice with Globo H-KLH and alpha-GalCer induced antibodies reactive with Globo H and SSEA3, suggesting that a Globo H-based vaccine will target tumor cells expressing Globo H or SSEA3. We next sought to reduce Globo H expression by siRNA targeting fucosyltransferase (FUT) 1 and 2, which mediate alpha-1,2 linkage of fucose to SSEA3 to generate Globo H. We showed both genes to be involved in the biosynthesis of Globo H. Moreover, FUT2 expression in BCSCs was significantly lower than in non-BCSCs harvested from a primary human breast cancer in NOD/SCID mouse, whereas FUT1 was slightly lower in BCSCs. Thus, the lower expression of Globo H in BCSCs may be attributed to less FUT2/FUT1, and to reduced SSEA3 in BCSCs compared with non-BCSCs. Our findings provide insight into further development of a Globo H-based vaccine and FUT1/FUT2-targeted therapy for breast cancer.
Chang
Wen-Wei
WW
Genomics Research Center, Academia Sinica, Taipei 115, Taiwan.
Lee
Chien Hsin
CH
Lee
Peishan
P
Lin
Juway
J
Hsu
Chun-Wei
CW
Hung
Jung-Tung
JT
Lin
Jin-Jin
JJ
Yu
Jyh-Cherng
JC
Shao
Li-en
LE
Yu
John
J
Wong
Chi-Huey
CH
Yu
Alice L
AL
D000818:Animals; D000906:Antibodies / Q000276:immunology; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000276:immunology / Q000378:metabolism*; D001943:Breast Neoplasms / Q000378:metabolism*; D002460:Cell Line; D005647:Fucosyltransferases / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D006028:Glycosphingolipids / Q000378:metabolism*; D006262:Health; D006801:Humans; D051379:Mice; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D055746:Stage-Specific Embryonic Antigens; D014612:Vaccines / Q000276:immunology; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D000906:Antibodies; D015295:Antigens, Tumor-Associated, Carbohydrate; D005647:Fucosyltransferases; D006028:Glycosphingolipids; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D055746:Stage-Specific Embryonic Antigens; D014612:Vaccines; D000097631:Galactoside 2-alpha-L-fucosyltransferase; C472765:Globo-H; C052924:stage-specific embryonic antigen-3