Ilisso Concetta Paola, Castellano Maria, Zappavigna Silvia, Lombardi Angela, Vitale Giovanni, Dicitore Alessandra, Cacciapuoti Giovanna, Caraglia Michele, Porcelli Marina
Department of Biochemistry, Biophysics and General Pathology, Second University of Naples, Via L. De Crecchio 7, 80138, Naples, Italy.
Endocrine. 2015 Sep;50(1):212-22. doi: 10.1007/s12020-014-0484-7. Epub 2015 Jan 11.
In this work, we have investigated the antiproliferative effect of AdoMet and Doxorubicin (Doxo), alone or in combination, on different breast cancer cell lines. For the evaluation of synergism, we have calculated the combination index (CI) by the Calcusyn software and we have evaluated the effects of the combination on apoptosis occurrence at FACS analysis in hormone-dependent CG5 cell line. We have found that AdoMet and Doxo given in combination were strongly synergistic in the hormone-dependent CG5 and MCF-7 human breast cancer cell line, as a CI50 < 0.5 was found after 72 h of treatment while the effect was only additive in hormone-independent MDA-MB 231 cells. On the basis of our results, we have selected a combination of AdoMet and Doxo, that was highly synergistic and we have found that the AdoMet in combination with Doxo increased apoptosis induced by Doxo alone, suggesting that the synergism on growth inhibition was largely due to apoptosis. Notably, the AdoMet/Doxo combination induced a significant activation of caspases 3, and 8, while no effect was found on caspase 9 cleavage. In contrast, no significant changes of the expression of cleaved caspase 8 and 9 were found in cells treated with AdoMet and Doxo alone. Moreover, the combination induced a significant increase of Fas and FasL expression. These results highlight the importance of the synergistic effect of AdoMet with Doxo in the regulation of hormone-dependent breast cancer cell proliferation and emphasize the anti-tumor activity of these molecules.
在本研究中，我们研究了腺苷甲硫氨酸（AdoMet）和阿霉素（Doxo）单独或联合使用对不同乳腺癌细胞系的抗增殖作用。为了评估协同作用，我们使用Calcusyn软件计算了联合指数（CI），并通过流式细胞术分析评估了联合用药对激素依赖性CG5细胞系凋亡发生的影响。我们发现，联合使用AdoMet和Doxo在激素依赖性CG5和MCF-7人乳腺癌细胞系中具有强烈的协同作用，因为在处理72小时后发现CI50<0.5，而在激素非依赖性MDA-MB 231细胞中该作用仅为相加作用。基于我们的结果，我们选择了一种高度协同的AdoMet和Doxo组合，并且发现AdoMet与Doxo联合使用可增加单独使用Doxo诱导的凋亡，这表明对生长抑制的协同作用很大程度上归因于凋亡。值得注意的是，AdoMet/Doxo组合诱导了半胱天冬酶3和8的显著激活，而对半胱天冬酶9的切割没有影响。相比之下，单独用AdoMet和Doxo处理的细胞中，切割的半胱天冬酶8和9的表达没有显著变化。此外，该组合诱导了Fas和FasL表达的显著增加。这些结果突出了AdoMet与Doxo协同作用在调节激素依赖性乳腺癌细胞增殖中的重要性，并强调了这些分子的抗肿瘤活性。
