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To determine whether pyrrolidine dithio- carbamate(PDTC) enhances TNFa-induced apoptosis in cultured breast cancer cells and explore the role of NF-kB in TNFa-induced apoptosis. Methods: Human breast cancer cell lines MCF-7 and MDA-MB-435s were treated with TNFa, PDTC and combination therapy. Cell survivals were determined by MTT assay. Apoptosis was detected by TUNEL and flow cytometry. NF-k B DNA binding activity was detected using electrophoresis mobility shift assay (EMSA). Western blots were performed to demonstrate IkBa(Inhibitor protein of nuclear factorkB) phosphorylation and degradation. Results: Cell growth was not suppressed by either TNFa(2000 U/ml or less) or PDTC alone. Both cell lines treated with TNFa (2000 U/ml) combined with PDTC(50 mmol/L) showed significant growth inhibition. PDTC inhibited TNFa-induced IkBa phosphorylation and degradation in both cell lines. EMSA showed that PDTC continuously inhibited TNFa induced NF-k B DNA binding activity. TNFa induced apoptosis (TUNEL) was increased significantly when both cells were pretreated with PDTC, and this was confirmed by Flow cytometry. Conclusion: PDTC enhances TNFa-induced apoptosis via inhibiting IkBa phosphorylation and degradation in human breast cancer cells. NF-k B protects against TNFa-induced apoptosis.
To determine whether pyrrolidine dithio-carbamate (PDTC) enhances TNFa-induced apoptosis in cultured breast cancer cells and explore the role of NF-kB in TNFa-induced apoptosis. Methods: Human breast cancer cell lines MCF-7 and MDA- MB-435s were treated with TNFa, PDTC and combination therapy. Cell survivals were determined by MTT assay. Apoptosis was detected by TUNEL and flow cytometry. NF-k B DNA binding activity was detected using electrophoresis mobility shift assay (EMSA). Western blots were performed to Results: Cell growth was not suppressed by either TNFa (2000 U / ml or less) or PDTC alone. Both cell lines treated with TNFa (2000 U / ml) combined with PDTC (50 mmol / L) showed significant growth inhibition. PDTC inhibited TNFa-induced IkBa phosphorylation and degradation in both cell lines. EMSA showed that PDTCusted TNFa induced NF-κB DNA binding activity. TNFa induced apoptosi (TUNEL) was increased significantly when both cells were pretreated with PDTC, and this was confirmed by Flow cytometry. Conclusion: PDTC enhances TNFa-induced apoptosis via inhibiting IkB phosphorylation and degradation in human breast cancer cells. NF-κ B protects against TNFa -induced apoptosis.