Control of metastatic properties of BL6 melanoma cells by H-2Kb gene: immunological and nonimmunological mechanisms.
The effect of class I H-2 antigen expression on the metastatic properties of BL6 melanoma cells was investigated. The BL6-8 clone isolated from the highly metastatic BL6 melanoma did not express H-2Kb gene. Following transfection with the H-2Kb gene, BL6-8 cells displayed a low metastatic potential in the immunocompetent as well as immunosuppressed (X-irradiated) or triple-immunodeficient mice with impaired T, B and natural killer (NK) cells function. The expression of H-2Kb gene and the low metastatic ability of transfected BL6 melanoma cells were associated with appearance of cell membrane soybean agglutinin (SBA) and Griffonia simplicifolia 1B4 (GS1B4) lectin-binding carbohydrates. These alterations in cell surface carbohydrates were found to be a result of reduction in sialylation of SBA binding sites and upregulation of the alpha 1.3 galactosyltransferase (alpha 1.3GT) gene. To assess the importance of H-2Kb-induced alterations in cell surface carbohydrates for metastasis formation, BL6-8 melanoma cells were transfected with H-2Kb gene without neor gene cotransfection and selected for adherence to SBA-lectin-conjugated agarose beads. The transfected clones that expressed SBA and GS1B4 lectin-binding carbohydrates were low metastatic. Further analysis of these clones showed that presence of SBA and GS1B4 lectin-binding carbohydrates rather than expression of H-2Kb molecules per se might be responsible for low metastatic potentials of H-2Kb-transfected cells in the immunocompromised mice. Studies of the possible mechanisms responsible for low metastatic ability of H-2Kb-transfected melanoma cells revealed that these cells displayed a reduced ability to adhere to murine pulmonary endothelial cells as well as to laminin and collagen IV. We hypothesized that the observed nonimmunological effects of H-2Kb gene in BL6 melanoma cells is a result of an interaction between the H-2Kb gene and B16 melanoma-specific ecotropic retrovirus. It results in inhibition of this retrovirus production with consecutive alteration in the expression of cellular genes controlling cell surface glycosylation and adhesion properties essential for the metastatic phenotype of BL6 melanoma.
Gorelik
E
E
Pittsburgh Cancer Institute, PA 15213.
Kim
M
M
Duty
L
L
Henion
T
T
Galili
U
U
D000818:Animals; D001483:Base Sequence; D002448:Cell Adhesion; D017931:DNA Primers / Q000737:chemistry; D004727:Endothelium / Q000166:cytology; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005700:Galactosyltransferases / Q000235:genetics; D015870:Gene Expression; D005802:Genes, MHC Class II; D006183:H-2 Antigens / Q000235:genetics*; D007153:Immunologic Deficiency Syndromes / Q000473:pathology; D037102:Lectins / Q000378:metabolism; D008545:Melanoma / Q000235:genetics / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis / Q000235:genetics*; D009439:Neuraminidase / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D016326:Extracellular Matrix Proteins; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D037102:Lectins; D012333:RNA, Messenger; D005700:Galactosyltransferases; C037959:beta-D-galactosyl-N-acetylglucosaminylglycopeptide alpha-1,3-galactosyltransferase; D009439:Neuraminidase