In vitro and in vivo effect of IFNalpha on B16F10 melanoma in two models: subcutaneous (C57BL6J mice) and lung metastasis (Swiss mice).

Interferon alpha tends to be the only agent used to treat melanoma. The objective of this study was to assess the effect of interferon alpha on the growth of the B16F10 melanoma, both in vitro and in vivo. We study the in vitro effect of interferon alpha (250,000, 500,000 and 1,000,000 IU/ml) on the B16F10 melanoma cell line (at 24, 48 and 72 h) and the in vivo effect in a subcutaneous (1x10(6) cells; 300,000 IU) and a pulmonary metastatic model (5x10(5) cells/lateral vein of the tail; 300,000, 600,000 and 1,200,000 IU). Necropsy included a morphological and immunohistochemical study (subcutaneous model), while the number of superficial lung metastases, implantation percentage and growth and invasion indices were calculated in the latter model. In vitro, interferon alpha decreased cell survival in a time- and dose-dependent manner; 250,000 IU/ml: 77% (24h), 80% (48 h) and 92% (72 h); 500,000 IU/ml: 62% (24h), 32% (48 h), 20% (72 h); 1,000,000 IU/ml: 41% (24h), 16% (48 h), 10% (72 h). In the subcutaneous model, it reduced tumor weights (77.74%) and cell proliferation (70.8%), and increased necrotic areas (8%) and inflammatory infiltrates (34.46%). Metastatic model: 300,000 IU reduced pleural nodules by 38.79%, implantation by 59.42%, growth by 43.48%, invasion by 25.06%; the corresponding figures for 600,000 and 1,200,000 IU were 38.79, 59.42, 43.48, 25.06%, and 65.55, 84.98, 56.52, 66.19%, respectively. Interferon alpha inhibited cell proliferation in all the models and had immunomodulatory (subcutaneous model) and antimetastatic (pulmonary metastatic model) effects in vivo.

Efectos del IFNα y del Lymphomyosot sobre el crecimiento del melanoma B16F10 «in vitro» e «in vivo». Tratamiento con IFNα y Lymphomyosot / Effects of IFNα and Lymphomyosot on «vitro» and «in vivo» B16F10 melanoma growth. Treatment with IFNα and Lymphomyosot

Introducción: En el tratamiento del melanoma se estánensayando distintas citoquinas, entre las que destaca elIFNα, no existiendo estudios experimentales con productoshomeopáticos. Material y métodos: «In vitro» la líneaB16F10 fue tratada durante 24, 48 y 72 horas con IFNα/10a 1.000.000 UI/ml y con Lymphomyosot 1/1 y 1/3 (v/v),cuantificando la viabilidad celular con el test del MTT. «Invivo» realizamos dos experimentos con 80 ratones machosa los que inyectamos 1X105 células B16F10 y tratamos con:I (PBS), II (IFNα), III (Lymphomyosot) y IV (IFNα másLymphomyosot), realizando el estudio morfológico. Resultados:«In vitro» el IFNα a altas concentraciones inhibió elcrecimiento celular y el Lymphomyosot no mostró efectoscitotóxicos. «In vivo» el IFNα redujo el índice de proliferacióncelular y la extensión de los infiltrados inflamatorios,mientras que el Lymphomyosot originó disminución significativade los pesos y necrosis tumorales. Conclusión: ElIFNα ha mostrado citotoxicidad en ambos modelos y elLymphomyosot ausencia de toxicidad y efecto antitumoralindirecto probablemente al aumentar la respuesta del huésped

Introduction: Several cytokines are being tested in thetreatment of melanoma. Among them IFNα should be highlighted,whilst no other experimental studies using homeopathicproducts are underway. Material & Methods: «Invitro»: the cell line B16F10 was treated at 24, 48 and 72hours with IFNα/10 at 1,000,000 IU/ml and withLymphomyosot 1/1 and 1/3 (v/v). Cell viability was quantifiedusing MTT test. «In vivo»: two experiments werecarried out on 80 male mice which were injected with 1x105B16F10 cells and treated with: I (PBS), II (IFNα), III(Lymphomyosot) and IV (IFNα plus Lymphomyosot). Amorphological study was also performed. Results: «Invitro»: at high concentrations of IFNα, cell growth wasinhibited and Lymphomyosot showed no cytotoxic effects.«In vivo»: IFNα reduced the cell-proliferation rate as wellas the extent of spread of inflammatory infiltrates, whilstLymphomyosot caused a significant tumor weight drop andnecrosis. Conclusion: IFNα displayed cytotoxicity in bothmodels whilst Lymphomyosot had an absence of toxicityand an indirect anti-tumor effect, probably due to an increasein the host's response

In vivo treatment of melanoma (B16F10) with a homeopathic agent and with a cytokine (IFN-alpha).

Among the numerous agents tested on melanoma, cytokines have attracted much attention over recent decades, in particular interferon-alpha (IFN-alpha). However, in a small number of experimental assays, homeopathic products have also been used. This study aimed to analyze the effects of INF-alpha and Lymphomyosot, administered individually or in combination, on the growth of B16F10 melanoma transplanted in C57BL/6J mice. Two experiments were performed using 72 young male mice, treated with 1 x 10(6) B16F10 cells and treated with phosphate-buffered saline (I), INF-alpha (II), Lymphomyosot (III), and both INF-alpha and Lymphomyosot (IV). Subsequent morphological and immunohistochemical studies were performed. All treatments produced a reduction in tumor weight with significant differences in those treated with INF-alpha and Lymphomyosot. INF-alpha reduced the cell proliferation index and the spread of inflammatory infiltrates and produced an increase in the extent of intratumoral necrosis. An antitumour effect was displayed by both agents, as was the cytotoxicity of INF-alpha and the immune response-stimulating effect of Lymphomyosot.