Potentiation of growth suppression and modulation of the antigenic phenotype in human melanoma cells by the combination of recombinant human fibroblast and immune interferons

Abstract

Administration of interferon as a single therapeutic regimen in cancer patients with various neoplasias has had only limited efficacy in ameliorating the negative clinical course of their disease. In the present study, we have evaluated the effect of recombinant human fibroblast (IFN beta) and immune (IFN gamma) interferon, alone and in combination, on growth, differentiation and the expression of class I and II histocompatibility locus antigens (HLA) and melanoma-associated antigens on the human melanoma cell line H0-1. The effect of combinations of interferons on the antigenic profile of human melanoma cells displaying different organ colonization and spontaneous metastatic potential in athymic nude mice was also determined. H0-1 cells were more sensitive to the antiproliferative activity of IFN beta than to IFN gamma and the combination of interferons resulted in a potentiation of growth suppression. The antiproliferative effect of both interferons was greater in later-passage than in earlier-passage H0-1 cells, possibly reflecting alterations in the evolving tumor cell population as a result of long-term in vitro propagation and/or the selective outgrowth of cells with an increased growth rate. The enhanced growth suppression observed in H0-1 cells treated with the combination of IFN beta plus IFN gamma was not associated with a significant increase in the level of melanin, a marker of melanoma differentiation, above that observed with either interferon used alone. IFN beta and IFN gamma differentially modulated the expression of class I and II HLA and melanoma-associated antigens in H0-1 cells and a series of melanoma cells with different organ colonization and metastatic potential, including MeWo, MeM 50-10, MeM 50-17, 3S5 and 70W. No consistent potentiation or antagonism in the expression of any specific antigen was observed in any of the melanoma cell lines exposed to the combination of interferons. The present study demonstrates that the combination of IFN beta plus IFN gamma can potentiate growth suppression in H0-1 human melanoma cells and that this effect is not associated with an increase in differentiation or a potentiation in antigenic modulation. In addition, no direct correlation between the expression of any specific antigen or its modulation by IFN beta or IFN gamma, alone or in combination, and organ colonization and metastatic potential in nude mice was observed in the different melanoma cell lines.

Genomic structure, chromosomal localization and expression profile of a novel melanoma differentiation associated (mda-7) gene with cancer specific growth suppressing and apoptosis inducing properties

“…The blot was stripped and then reprobed with a 32 P-labeled GAPDH probe. FM516 is an SV40-immortalized normal melanocyte cell line (FM516-SV); WM35 is an early RGP primary melanoma cell line; WM278 is an early VGP primary melanoma cell line; WM239, FO-1, C8161, MeWo, 3S5 and 70W are metastatic melanoma cell lines; SK-MEL p53 mt is a metastatic melanoma cell line with a con®rmed mutant p53 genotype; and SK-MEL p53 wt is a metastatic melanoma cell line with a con®rmed wild-type p53 genotype having a mutant p53 gene by itself can prevent mda-7 expression, since MeWo and its two subclones, 3S5 displaying a reduction in metastatic competence and 70W exhibiting enhanced metastatic potential (Kerbel and Man, 1984;Graham et al, 1991), which contain one mutant and one wild-type p53 allele are readily inducible for mda-7 expression following IFN-b+MEZ treatment. These studies document de novo expression of mda-7 in cultured normal human melanocytes, as observed with patient-derived melanocytes, and they provide additional support for the idea that mda-7 may function as a negative regulator of melanoma progression (Jiang et al, 1995b).…”

“…Treatment of human melanoma cells with IFNb+MEZ results in profound physiological changes, including an irreversible loss in proliferative potential, suppression of oncogenic potential in athymic nude mice, altered cell surface antigenicity, temporal alterations in gene expression and induction of terminal di erentiation (Fisher et al, 1985;Graham et al, 1991;Jiang et al, , 1994Jiang et al, , 1995aJiang et al, ,b, 2000Huang et al, 1999a,b;Kang et al, 2001;Leszczyniecka et al, 2001). To obtain insights into this process we have begun to de®ne the spectrum of gene expression changes occurring as a consequence of this combination treatment in human melanoma cells using several molecular approaches.…”

“…C8161 metastatic melanoma cells were obtained from Dr D Welch (University of Pennsylvania) (Jiang et al, 1995a). Dr RS Kerbel provided the MeWo cell line and its reduced metastatic variant 3S5 and highly metastatic variant 70W (Kerbel and Man, 1984;Graham et al, 1991). SK-MEL p53 mt and SK-MEL p53 wt were provided by Dr A Albino (American Health Foundation, NY, USA).…”

The cancer growth suppressing gene mda-7 induces apoptosis selectively in human melanoma cells

“…HO-1 and MeWo are established human melanotic melanoma cell lines and FO-1 is an established human amelanotic melanoma cell line isolated from separate patient-derived metastatic lesions (Giovanella et al, 1976;Kerbel et al, 1994;Fisher et al, 1985;Graham et al, 1991). WM35 was derived from an RGP primary human melanoma (Herlyn, 1990).…”

Down-regulation of Myc as a Potential Target for Growth Arrest Induced by Human Polynucleotide Phosphorylase (hPNPase) in Human Melanoma Cells

“…The other is the pathological process of neoplastic transformation characterized by uncontrolled proliferation and de-differentiation. Treatment of HO-1 metastatic human melanoma cells with fibroblast interferon (IFN-␤) 1 and the protein kinase C activator mezerein (MEZ) induces irreversible growth arrest and terminal differentiation characterized by changes in cell morphology, increase in melanin synthesis, modifications in gene expression, and alterations in surface antigen expression (1)(2)(3)(4)(5). Replicative or cellular senescence, a process leading to irreversible arrest of cell division, was first described in cultures of human fibroblasts that lost the ability to divide upon continuous subcultures (6).…”