Department of Pharmacodynamics, Medical University of Bialystok, Mickiewicza 2C, 15-222 Białystok, Poland.
Pharmacol Rep. 2010 Sep-Oct;62(5):926-37. doi: 10.1016/s1734-1140(10)70353-4.
Supplementation of recombinant human erythropoietin (rHuEpo) is one of the methods for the treatment of anemia. The influence of rHuEpo on proliferation or clonogenic growth of cancer cells is not clear and some of the published results are conflicting. The aim of this work was to study the effect of rHuEpo on colon cancer cells when given alone or in combination with cytostatics. Human colon adenocarcinoma cells (DLD-1) were cultured in medium with rHuEpo, 5-fluorouracil (5-FU) and an active metabolite of irinotecan (SN-38). Cell viability was determined using a hematocytometer and 0.4% (w/v) trypan blue dye. Cell proliferation was measured by the MTT assay. Expression of EpoR, Bax, Bcl-2 and Akt1 protein was assessed by Western blot. The results of this study indicate a dose-dependent inhibitory effect of rHuEpo on DLD-1 cell growth and proliferation. Moreover, the combined treatment of rHuEpo and cytotoxic agents such as 5-FU and SN-38 increases the antitumor action, which is indicated by decreases in proliferation in the MTT test, cell numbers and DNA synthesis. We found a significant increase in EpoR, Bcl-2 and Akt1 protein expression in all cells grown in medium containing 3 IU of rHuEpo. We observed that EpoR is constitutively expressed in DLD-1 cells. Our results indicate that rHuEpo acts via EpoR to directly inhibit DLD-1 cell growth and indirectly modulate the cytostatics effects of 5-FU and SN-38.
重组人促红细胞生成素（rHuEpo）的补充是治疗贫血的方法之一。rHuEpo 对癌细胞的增殖或集落形成生长的影响尚不清楚，一些已发表的结果相互矛盾。本工作旨在研究 rHuEpo 单独或与细胞毒药物联合应用对结肠癌细胞的影响。人结肠腺癌细胞（DLD-1）在含有 rHuEpo、5-氟尿嘧啶（5-FU）和伊立替康活性代谢物（SN-38）的培养基中培养。使用血细胞计数器和 0.4%（w/v）台盼蓝染料测定细胞活力。通过 MTT 测定法测量细胞增殖。通过 Western blot 评估 EpoR、Bax、Bcl-2 和 Akt1 蛋白的表达。本研究的结果表明 rHuEpo 对 DLD-1 细胞生长和增殖具有剂量依赖性抑制作用。此外，rHuEpo 与细胞毒药物如 5-FU 和 SN-38 的联合治疗增加了抗肿瘤作用，这表现在 MTT 试验、细胞数量和 DNA 合成的增殖减少。我们发现，在含有 3IU rHuEpo 的培养基中生长的所有细胞中，EpoR、Bcl-2 和 Akt1 蛋白的表达均显著增加。我们观察到 EpoR 在 DLD-1 细胞中持续表达。我们的结果表明，rHuEpo 通过 EpoR 直接抑制 DLD-1 细胞生长，并间接调节 5-FU 和 SN-38 的细胞毒作用。
