N-RAS mutations and susceptibility to lymphokine-activated killer (LAK) cells in human melanoma.
RAS oncogene expression has been reported to affect several biological features of rodent tumours cells, including lysability by activated natural killer cells. In order to examine whether expression of mutated RAS genes in human melanoma cells alters their susceptibility to lysis by LAK cells, seven melanoma lines were assessed for the presence of Ki- and N-RAS genes bearing all possible mutations at codons 12, 13 and 61. A panel of 21 clones deriving from the metastatic lesion Me665/2, which had a Gln-->Arg substitution at codon 61 of N-RAS (N-RAS/61+), were also examined. Melanoma cells and clones were used as targets of allogeneic LAK in a 4-h 51Cr-release assay. LAK showed a higher lysis on melanoma lines and clones harbouring a mutated RAS compared with counterparts bearing no RAS mutations. In addition, LAK-mediated lysis drastically decreased on Me665/2 sublines progressively selected by exposure to LAK. This loss was paralleled by a reduction or even disappearance of N-RAS/61+ mRNA signal in Me665/2 sublines. To evaluate whether N-RAS could directly modulate LAK susceptibility to lysis, N-RAS/61+ gene was transfected in two N-RAS wild type (N-RAS/61-) 665/2 melanoma clones by a cosmid vector. In contrast to the high lysability of melanoma cells constitutively expressing the mutationally active N-RAS oncogene, N-RAS/61+ transfectants did not show a consistent high lysability by LAK, compared with some control pSV2neo transfectants.(ABSTRACT TRUNCATED AT 250 WORDS)
Division of Experimental Oncology D, Istituto Nazionale Tumori, Milan, Italy.
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R
Lupetti
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Sensi
R
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Mortarini
A
A
Anichini
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Clemente
G
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Parmiani
D001483:Base Sequence; D002999:Clone Cells; D015972:Gene Expression Regulation, Neoplastic; D011905:Genes, ras / Q000235:genetics*; D006801:Humans; D016219:Immunotherapy, Adoptive; D015979:Killer Cells, Lymphokine-Activated; D008213:Lymphocyte Activation; D008545:Melanoma / Q000235:genetics* / Q000276:immunology / Q000628:therapy*; D008969:Molecular Sequence Data; D009154:Mutation; D014162:Transfection; D014407:Tumor Cells, Cultured