Eg5 inhibitor, a novel potent targeted therapy, induces cell apoptosis in renal cell carcinoma
Eg5 inhibitor, a novel potent targeted therapy, induces cell apoptosis in renal cell carcinoma
复制标题
DOI：
10.1007/s13277-014-2022-x
复制
发表时间：
2014-08-01
期刊：
影响因子：
--
通讯作者：
Ding, Kejia
中科院分区：
文献类型：
--
作者：
Ding, Sentai;Zhao, Zuohui;Ding, Kejia
Eg 5是有丝分裂的关键，在多种恶性肿瘤中过表达，现已被确定为癌症治疗的一个有前途的靶点。然而，Eg 5抑制剂在肾细胞癌（RCC）中的抗癌活性仍然是一个悬而未决的问题。在本文中，我们首次评估了S-（甲氧基三苯甲基）-l-半胱氨酸（S（MeO）TLC）在体外和体内阻断Eg 5在RCC中的治疗益处。在临床组织样品和各种肾细胞系（包括293 T、786-0和OS-RC-2）中检查Eg 5的表达。Eg 5抑制剂（S）-三苯甲基-1-半胱氨酸（STLC）和S（MeO）TLC）的抗增殖活性通过细胞活力测定来评价。应用Hoechst核染色和流式细胞术的凋亡测定来研究S（MeO）TLC的功效，S（MeO）TLC比STLC更有效。采用免疫荧光法研究其可能的作用机制。此外，通过使用皮下异种移植模型进行体内研究，其用于确认其作为潜在抗肿瘤药物的作用。在肾细胞系和肾细胞癌组织中检测到Eg 5的表达，其在正常肾脏样品中表达较低。STLC和S（MeO）TLC在72 h表现出最佳的抗增殖活性，S（MeO）TLC处理24 h后细胞呈现典型的单星梭形表型，48 h后出现凋亡细胞。在体内，S（MeO）TLC有效抑制皮下异种移植模型中的肿瘤生长。在体外和体内，抑制Eg 5抑制RCC的增殖。所有这些发现共同证明S（MeO）TLC，一种有效的Eg 5抑制剂，是用于治疗RCC的有希望的抗癌剂。
Eg5 is critical for mitosis and overexpressed in various malignant tumors, which has now been identified as a promising target in cancer therapy. However, the anti-cancer activity of Eg5 inhibitor in renal cell carcinoma (RCC) remains an open issue. In this paper, we evaluated, for the first time, the therapeutic benefit of blocking Eg5 by S-(methoxytrityl)-l-cysteine (S(MeO)TLC) in RCC both in vitro and vivo. The expression of Eg5 was examined in clinical tissue samples and various kidney cell lines, including 293T, 786-0, and OS-RC-2. The anti-proliferative activity of Eg5 inhibitors, (S)-trityl-l-cysteine (STLC) and S(MeO)TLC, was evaluated by a cell viability assay. An apoptosis assay with Hoechst nuclear staining and flow cytometry was applied to investigate the efficacy of the S(MeO)TLC, which is more potent than STLC. Immunofluorescence was used to research the possible mechanism. Furthermore, in vivo studies were performed by using subcutaneous xenograft models, which were used to confirm its role as a potential anti-neoplastic drug. The Eg5 expression was detected in kidney cell lines and RCC tissues, which was low in normal kidney samples. STLC and S(MeO)TLC exhibited their optimal anti-proliferative activity in 72 h, and cells treated with S(MeO)TLC presented characteristic monoastral spindle phenotype in 24 h and apoptotic cells in 48 h. In vivo, S(MeO)TLC effectively suppressed tumor growth in subcutaneous xenograft models. Inhibition of Eg5 represses the proliferation of RCC in vitro and in vivo. All these findings collectively demonstrate that S(MeO)TLC, a potent Eg5 inhibitor, is a promising anti-cancer agent for the treatment of RCC.