Different levels of TGFbeta, IL-10, IFNgamma and gelatinase A occur in experimental white and black metastases induced by bryostatin 1 or by phorbol ester-treated BL6T murine melanoma cells

Clin Exp Metastasis. 2000;18(5):361-9. doi: 10.1023/a:1010913907830.

Abstract

Bryostatin 1 and phorbol esters reduce the intracellular melanin level in high metastatic overexpressing nPKCdelta BL6 (BL6T) cells, thereby inducing white experimental metastasis in syngeneic mice. We evaluate here the possible differences between white and black metastases induced by both treatments on the proliferative and metastatic potential as well as on the expression of some cytokines involved in the metastatic process such as TGFbeta, IL-10 and IFNgamma. The level of expression of gelatinase A is also considered. White and black metastases induced after the injection of bryostatin 1- or phorbol ester-treated cells into the tail vein of syngenic mice were isolated and analysed for the levels of LDH usually used as markers of cytotoxicity, for the levels of cytokines and gelatinase A or dissociated and cultured in vitro for a few passages. The cultured cells were analysed in vitro for the proliferative capacity and the melanin synthesis. The same cells were also re-injected into syngeneic mice and the number of experimental metastases were counted after 17 days or injected with matrigel in order to quantify the proliferative capacity in vivo. The results show only one significant difference between bryostatin I and phorbol ester, namely the cells obtained from white bryostatin 1-treated cells return to a black phenotype after a few passages in culture. This suggests that PKC mediates many of the biological effects of bryostatin 1 but that its effect is lost in vitro. On the other hand, white and black metastases (at least for metastases induced by BL6T cells treated with phorbol ester) do appear significantly different. In vivo white metastases show lower levels of LDH, lower levels of proliferative capacity into matrigel, higher levels of TGFbeta and IFNgamma and, when re-injected into syngeneic mice, give big black metastases. Therefore, in murine melanoma cells, the treatment with bryostatin I induces the appearance of a white population expressing different levels of TGFbeta, IFNgamma, IL-10 and gelatinase A. Such a white population is more difficult to diagnose and is capable of turning into a more aggressive phenotype under suitable environmental conditions.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Antineoplastic Agents / adverse effects
  • Bryostatins
  • Interferon-gamma / genetics
  • Interferon-gamma / metabolism*
  • Interleukin-10 / genetics
  • Interleukin-10 / metabolism*
  • L-Lactate Dehydrogenase / metabolism
  • Lactones / adverse effects
  • Lung Neoplasms / chemically induced
  • Lung Neoplasms / metabolism
  • Lung Neoplasms / pathology*
  • Macrolides
  • Matrix Metalloproteinase 2 / drug effects
  • Matrix Metalloproteinase 2 / genetics
  • Matrix Metalloproteinase 2 / metabolism*
  • Melanoma / metabolism
  • Melanoma / secondary*
  • Mice
  • Mice, Inbred C57BL
  • Protein Kinase C / drug effects
  • Protein Kinase C / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tetradecanoylphorbol Acetate / toxicity
  • Transforming Growth Factor beta / genetics
  • Transforming Growth Factor beta / metabolism*

Substances

  • Antineoplastic Agents
  • Bryostatins
  • Lactones
  • Macrolides
  • Transforming Growth Factor beta
  • Interleukin-10
  • bryostatin 1
  • Interferon-gamma
  • L-Lactate Dehydrogenase
  • Protein Kinase C
  • Matrix Metalloproteinase 2
  • Tetradecanoylphorbol Acetate