The Akt-inhibitor Erufosine induces apoptotic cell death in prostate cancer cells and increases the short term effects of ionizing radiation.
The Akt-inhibitor Erufosine induces apoptotic cell death in prostate cancer cells and increases the short term effects of ionizing radiation.
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DOI：
10.1186/1748-717x-5-108
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发表时间：
2010-11-16
期刊：
影响因子：
--
通讯作者：
Jendrossek V
中科院分区：
文献类型：
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作者：
Rudner J;Ruiner CE;Handrick R;Eibl HJ;Belka C;Jendrossek V
磷脂酰肌醇-3-激酶（PI 3 K）/Akt通路在前列腺癌中经常失调，并与肿瘤转化、恶性进展以及对经典化疗和放疗的耐药性增强相关。因此，它是一个有前途的治疗干预的目标。在本研究中，分析了Akt抑制剂Erufosine（ErPC 3）在前列腺癌细胞中的细胞毒性作用，并与PI 3 K抑制剂LY 294002的细胞毒性进行了比较。此外，检查了Akt抑制剂和电离辐射在前列腺癌细胞中的组合治疗的功效。前列腺癌细胞系PC 3、DU 145和LNCaP用ErPC 3（1-100 μM）、LY 294002（25-100 μM）处理、辐照（0-10戈伊）或进行组合处理。通过WST-1测定法测定细胞活力。在低渗柠檬酸盐缓冲液中用碘化丙啶染色后，通过流式细胞术分析细胞凋亡诱导，并通过使用抗胱天蛋白酶-3及其底物PARP的抗体的Western印迹分析。通过蛋白质印迹分析检测Akt活性以及对Bcl-2家族成员和参与细胞凋亡调节的关键下游效应物表达的调节。Akt抑制剂ErPC 3在前列腺癌细胞中发挥抗肿瘤作用，但效力不同。ErPC 3的抗肿瘤作用与磷酸丝氨酸473-Akt水平降低和细胞凋亡诱导有关。PC 3和LNCaP前列腺癌细胞也对PI 3 K抑制剂LY 294002的治疗敏感。然而，ErPC 3敏感的PC 3-细胞对LY 294002的敏感性低于ErPC 3难治的LNCaP细胞。虽然这两种细胞系在很大程度上抵抗辐射诱导的细胞凋亡，这两种细胞系显示出更高水平的凋亡细胞死亡时，ErPC 3与放疗相结合。我们的数据表明，在两种前列腺癌细胞系中，组成性Akt激活和存活由不同的分子机制控制-一种对Akt抑制剂ErPC 3敏感，一种对PI 3 K抑制剂LY 294002更敏感。我们的研究结果强调了定义预测性生物标志物的重要性，这些生物标志物允许选择可能受益于特定信号转导修饰剂治疗的患者。
The phosphatidylinositol-3-kinase (PI3K)/Akt pathway is frequently deregulated in prostate cancer and associated with neoplastic transformation, malignant progression, and enhanced resistance to classical chemotherapy and radiotherapy. Thus, it is a promising target for therapeutic intervention. In the present study, the cytotoxic action of the Akt inhibitor Erufosine (ErPC3) was analyzed in prostate cancer cells and compared to the cytotoxicity of the PI3K inhibitor LY294002. Moreover, the efficacy of combined treatment with Akt inhibitors and ionizing radiation in prostate cancer cells was examined. Prostate cancer cell lines PC3, DU145, and LNCaP were treated with ErPC3 (1-100 µM), LY294002 (25-100 µM), irradiated (0-10 Gy), or subjected to combined treatments. Cell viability was determined by the WST-1 assay. Apoptosis induction was analyzed by flow cytometry after staining with propidium iodide in a hypotonic citrate buffer, and by Western blotting using antibodies against caspase-3 and its substrate PARP. Akt activity and regulation of the expression of Bcl-2 family members and key downstream effectors involved in apoptosis regulation were examined by Western blot analysis. The Akt inhibitor ErPC3 exerted anti-neoplastic effects in prostate cancer cells, however with different potency. The anti-neoplastic action of ErPC3 was associated with reduced phosphoserine 473-Akt levels and induction of apoptosis. PC3 and LNCaP prostate cancer cells were also sensitive to treatment with the PI3K inhibitor LY294002. However, the ErPC3-sensitive PC3-cells were less susceptible to LY294002 than the ErPC3-refractory LNCaP cells. Although both cell lines were largely resistant to radiation-induced apoptosis, both cell lines showed higher levels of apoptotic cell death when ErPC3 was combined with radiotherapy. Our data suggest that constitutive Akt activation and survival are controlled by different different molecular mechanisms in the two prostate cancer cell lines - one which is sensitive to the Akt-inhibitor ErPC3 and one which is more sensitive to the PI3K-inhibitor LY294002. Our findings underline the importance for the definition of predictive biomarkers that allow the selection patients that may benefit from the treatment with a specific signal transduction modifier.