Bcl-2 and glutathione depletion sensitizes B16 melanoma to combination therapy and eliminates metastatic disease

Clin Cancer Res. 2007 May 1;13(9):2658-66. doi: 10.1158/1078-0432.CCR-06-2642.

Abstract

Purpose: Advanced melanoma resists all current therapies, and metastases in the liver are particularly problematic. Prevalent resistance factors include elevated glutathione (GSH) and increased expression of bcl-2 in melanoma cells. GSH has pleiotropic effects promoting cell growth and broad resistance to therapy, whereas Bcl-2 inhibits the activation of apoptosis and contributes to elevation of GSH. This study determined the in vivo efficacy of combination therapies administered while GSH and Bcl-2 were individually and simultaneously decreased in metastatic melanoma lesions.

Experimental design: Highly metastatic murine B16 melanoma (B16M-F10) cells have elevated levels of both GSH and Bcl-2. B16M-F10 cells were injected i.v. to establish metastatic lesions in vivo. GSH was decreased using an L-glutamine--enriched diet and administration of verapamil and acivicin, whereas Bcl-2 was reduced using oligodeoxynucleotide G3139. Paclitaxel, X-rays, tumor necrosis factor-alpha, and IFN-gamma were administered as a combination therapy.

Results: Metastatic cells were isolated from liver to confirm the depletion of GSH and Bcl-2 in vivo. Reduction of Bcl-2 and GSH, combined with partial therapies, decreased the number and volume of invasive B16M-F10 foci in liver by up to 99% (P<0.01). The full combination of paclitaxel, X-rays, and cytokines eliminated B16M-F10 cells from liver and all other systemic disease, leading to long-term survival (>120 days) without recurrence in 90% of mice receiving the full therapy. Toxicity was manageable; the mice recovered quickly, and hematology and clinical chemistry data were representative of accepted clinical toxicities.

Conclusions: Our results suggest a new strategy to induce regression of late-stage metastatic melanoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents, Phytogenic / therapeutic use*
  • Combined Modality Therapy
  • Glutamine / administration & dosage
  • Glutathione / antagonists & inhibitors*
  • Melanoma, Experimental / drug therapy
  • Melanoma, Experimental / radiotherapy
  • Melanoma, Experimental / therapy*
  • Mice
  • Neoplasm Metastasis
  • Paclitaxel / therapeutic use*
  • Proto-Oncogene Proteins c-bcl-2 / antagonists & inhibitors*
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Skin Neoplasms / drug therapy
  • Skin Neoplasms / radiotherapy
  • Skin Neoplasms / therapy*
  • Thionucleotides / pharmacology
  • Treatment Outcome
  • Tumor Necrosis Factor-alpha / therapeutic use*
  • X-Rays

Substances

  • Antineoplastic Agents, Phytogenic
  • Proto-Oncogene Proteins c-bcl-2
  • Thionucleotides
  • Tumor Necrosis Factor-alpha
  • Glutamine
  • oblimersen
  • Glutathione
  • Paclitaxel