Eg5在肾癌进展中的作用及靶向抑制Eg5治疗耐药性肾癌的效果和机制

Renal cell carcinoma (RCC) is the leading cause of death in urogenital malignancies, and RCC is resistant to chemotherapy and radiotherapy and even the existing targeted therapy. Thus, the prognosis for patients with RCC is dismal because of high recurrence and metastasis. In addition, the mechanism of RCC progression is unclear. .The mitotic kinesin Eg5, known as kinesin spindle protein (KSP), which plays a critical role in the bipolar spindle formation and separation of duplicated chromosome during the early phase of mitosis, has been identified as an attractive anticancer target. Present studies show that Eg5 is closely related to tumor formation, progression and prognosis. Moreover, our previous studies confirmed that. Nonetheless, the regulatory mechanism of Eg5 on tumor proliferation, metastasis and invasion is rarely understood. And no study is reported on targeted Eg5 inhibition on RCC to date. Thus, the present study is to explore the role of Eg5 in RCC progression and clarify its regulatory mechanism by several experiment skills, and elucidate the therapeutic effect and mechanism of siRNA interfere and Eg5 inhibitors on RCC treatment. It's the first time to examine the therapeutic effect of Eg5 inhibitors in vivo by establishing the drug-resistance RCC animal model in the present study. Additionally, the present study aim to investigate the relationship between Eg5 expression and the prognosis in RCC patients. We truly hope our present study can provide effective theoretical basis on the mechanism of RCC progression and new approaches on targeted therapy of advanced RCC, which could provide a preferably prognosis for the patients with advanced RCC.

肾癌是致死率最高的泌尿系肿瘤，对放化疗均不敏感，甚至对现有靶向治疗也已出现耐药，并因术后复发、转移率高而预后很差。有丝分裂驱动蛋白Eg5在细胞增殖中起关键作用，已成为抗癌靶向治疗的研究热点，既有研究发现Eg5与肿瘤发生、进展及预后密切相关，我们的前期研究也对此给与了印证。但Eg5在肾癌进展即增殖、迁徙及侵袭中的调控作用仍不明确，迄今也没有靶向抑制Eg5治疗肾癌的报道。因此，本课题拟通过多种实验技术证实Eg5在肾癌进展中的作用并探讨其信号传导调控机制，阐明小分子靶向抑制剂及siRNA抑制Eg5功能治疗肾癌的效果和机制。通过建立现有靶向药物耐药肾癌动物模型，首次探讨靶向抑制Eg5治疗耐药肾癌的效果。此外，研究还通过分析临床肾癌患者资料，进一步探讨Eg5表达与肾癌临床进展、术后预后之间的关系。希望为肾癌进展机制以及靶向治疗提供新的有效理论依据，最终为晚期肾癌患者提供新的治疗思路和更好预后。

肾癌是致死率最高的泌尿系肿瘤，对放化疗均不敏感，甚至对现有靶向治疗也已出现耐药，并因术后复发、转移率高而预后很差。有丝分裂驱动蛋白Eg5 在细胞增殖中起关键作用，是近年来抗癌靶向治疗的研究热点，且研究提示Eg5 与肿瘤发生、进展及预后密切相关，我们的前期其他肿瘤中的研究结果也对此给与了印证，但在此之前尚缺乏肾癌中的相关研究。本课题针对Eg5在肾癌发生发展中的机制，以及以Eg5作为靶点的治疗效果进行了多方面系统的研究。. 临床研究通过免疫组化检测了肾癌标本Eg5表达与患者肿瘤进展预后的关系，结果提示Eg5与肾癌分期分级有关且是独立的预后因素，高表达患者预后较差，其中部分患者目前还在进行临床靶向治疗的随访，该结果则需要长时间随访获得。体外细胞研究第一部分提示Eg5高表达具有增加肾癌细胞增殖、进展的作用，研究通过质粒稳定上调表达Eg5细胞株，细胞粘附和侵袭能力增加，相关分子指标结果也得到验证，其中Eg5上调可上调HIF-1α及靶基因。以此为基础课题通过细胞实验证实siRNA 和小分子靶向抑制剂S (MeO)TLC两种方式抑制Eg5 功能可以使肾癌细胞有丝分裂停滞并因一部通过Caspase 途径凋亡。进一步的体内动物实验也证实Eg5靶向抑制剂S (MeO)TLC可以有效的治疗肾癌；动物研究中成功建立了舒尼替尼耐药肾癌模型，结果发现S (MeO)TLC对现有靶向药物舒尼替尼耐药的肾癌也有相似治疗效果，且体内动物实验中对治疗后的肿瘤进行了HE染色提示肿瘤细胞有丝分裂停滞后的单星细胞。. 本课题的研究结果不但提示了Eg5在肾癌进展中的作用和机制，丰富了肾癌进展分子生物学机制研究，更重要的是通过靶向抑制Eg5功能治疗肾癌的结果对肾癌靶向治疗提供新的靶点和理论依据，最终希望为晚期肾癌患者提供更好预后。