Bcl-2 and Mn-SOD Antisense Oligodeoxynucleotides and a Glutamine-enriched Diet Facilitate Elimination of Highly Resistant B16 Melanoma Cells by Tumor Necrosis Factor-α and Chemotherapy

摘要: Mitochondrial glutathione (mtGSH) depletion increases sensitivity of Bcl-2-overexpressing B16 melanoma (B16M)-F10 cells (high metastatic potential) to tumor necrosis factor-alpha (TNF-alpha)-induced oxidative stress and death in vitro. In vivo, mtGSH B16M-F10 was achieved by feeding mice (where the grew as a solid footpad) with an L-glutamine (L-Gln)-enriched diet, which promoted increase glutaminase activity, accumulation cytosolic L-glutamate, competitive inhibition GSH transport into mitochondria. L-Gln-adapted cells, isolated using anti-Met-72 monoclonal antibodies flow cytometry-coupled cell sorting, were injected portal vein produce hepatic metastases. l-Gln-adapted invasive (iB16M-Gln+) from liver same methodology treated TNF-alpha antisense Bcl-2 oligodeoxynucleotide, viability decreased approximately 12%. iB16M-Gln+ associated increased generation O2*- H2O2, opening mitochondrial permeability transition pore complex, release proapoptotic molecular signals. Activation mechanisms prevented ester-induced replenishment. The stress-resistant survivors showed adaptive response that includes overexpression manganese-containing superoxide dismutase (Mn-SOD) catalase activities. By treating double anti- therapy (Bcl-2 SOD2 oligodeoxynucleotides) TNF-alpha, survival 1%. Chemotherapy (taxol plus daunorubicin) easily removed this minimum percentage survivors. This contribution identifies critical molecules can be sequentially targeted facilitate elimination highly resistant cells.