LACK OF HLA CLASS-I ANTIGEN EXPRESSION BY MELANOMA-CELLS SK-MEL-33 CAUSED BY A READING FRAMESHIFT IN BETA-2-MICROGLOBULIN MESSENGER-RNA

The lack of HLA class I antigen expression by the melanoma cell line SK-MEL-33 is caused by a unique lesion in beta2-microglobulin (beta2-mu). Sequencing of beta2-mu mRNA detected a guanosine deletion at position 323 in codon 76 that causes a frameshift with a subsequent introduction of a stop codon at a position 54 base upstream of the normal position of the stop codon in the message. The loss of 18 amino acids and the change of 6 amino acids, including a cysteine at position 80 in the carboxy terminus of beta2-mu, are likely to cause marked changes in the structure of the polypeptide. The latter may account for the inability of beta2-mu to associate with HLA class I heavy chains and for its lack of reactivity with the anti-beta2-mu mAb tested. HLA class I antigen expression on SK-MEL-33 cells was reconstituted after transfection with a wild-type B2m gene, therefore indicating that the abnormality of endogenous B2m gene is the only mechanism underlying lack of HLA class I antigen expression by SK-MEL-33 cells. The guanosine deletion in B2M gene was detected also in the melanoma tissue from which SK-MEL-33 cells had originated. Therefore, the molecular lesion identified in the SK-MEL-33 melanoma cell line is not caused by a mutation acquired during growth in vitro but is likely to reflect a somatic mutation during tumor progression.