NDUFA4L2抑制ROS形成参与肾癌发生发展及其分子靶向治疗的研究

Renal cell carcinoma (RCC) is not sensitive to chemoradiotherapy, and targeted therapy has become first-line treatment in advanced RCC. Therefore, looking for a new target for RCC and then developing a targeted drug therapy are significant. We found NDUFA4L2 was a renal cell carcinoma specific highly expression gene with low expression or no expression in adjacent tissues of RCC and other tumor through bioinformatics. In addition, the data of the TCGA support that the expression of NDUFA4L2 is significantly related with the tumor size, histologic grade and prognosis in RCC. The expression of NDUFA4L2 in clinical samples were detected by real-time PCR, Western blot and tissue microarray immunohistochemistry staining, which was further supported the result of bioinformatical analysis. This project intends to study the function of NDUFA4L2 through interference and overexpression technology in vivo and in vitro, and analyze the molecular mechanism of NDUFA4L2 in the occurrence and development of RCC via inhibiting ROS generation. what's more, This research want to explore if complex I inhibitor rotenone could affect the NDUFAL2 function and become a targeted therapeutic agent for RCC. This project intend to validate that NDUFA4L2 as RCC-specific markers could have important diagnostic or progrostic value in RCC. The study also try to uncover the role of NDUFA4L2 in the occurrence and development of RCC and explore if it can be used as a potential therapeutic target for the treatment of RCC, which may offer new way for the diagnosis and prognosis of RCC.

肾癌对放化疗不敏感，靶向治疗已成为晚期肾癌一线治疗。因此,寻找肾癌新靶点及研发相应靶向药物具有重要意义。我们通过生物信息学发现NDUFA4L2为肾癌特异高表达基因。另外TCGA的肾癌数据支持该基因与肿瘤大小，分级及预后明显相关。我们在临床标本中通过real-time PCR、Western blot以及组织芯片免疫组化进一步验证了生物信息学结果。本项目拟通过干扰、过表达等技术在体内外研究NDUFA4L2的功能，以及分析该基因通过抑制ROS形成参与肾癌发生发展的分子机制。本课题也将探讨复合体I抑制剂鱼藤酮能否影响NDUFA4L2的功能从而作为肾癌靶向治疗药物。本项目欲验证NDUFA4L2作为肾癌特异性肿瘤标志物，在肾癌的诊断、预后中是否具有重要的价值。同时去揭示NDUFA4L2在肾癌发生发展的作用机制以及探讨其该基因能否作为肾癌靶向治疗一个潜在药物靶点，从而为肾癌的诊断治疗提供新思路。

早期肾癌可通过手术切除，患者5年生存率 95%以上，但是晚期转移性肾癌5年生存率低于 10%。鉴于肾癌对放化疗不敏感，靶向治疗已成为晚期肾癌一线治疗。尽管苹果酸舒尼替尼作为晚期肾癌的首选治疗，但其客观反应率仍不到1/3。因此，寻找肾癌新靶点及研发相应靶向药物具有重要意义。我们通过生物信息学发现NDUFA4L2为肾癌特异高表达基因。另外TCGA数据库也表明该基因与肾透明细胞癌肿瘤大小，临床分期，病理分级及患者预后明显相关。我们在临床标本中通过real-time PCR、Western blot以及组织芯片免疫组化进一步验证了生物信息学结果。此外本课题通过干扰肾癌细胞系NDUFA4L2的表达，以及体内移植瘤试验证明NDUFA4L2对于肾癌的发生发展起着重要的作用。同时本研究发现通过低氧处理，细胞系HIF1及HIF2表达量明显升高，而NDUFA4L2的表达量也随之增加。提示NDUFA4L2是通过HIF的调控而介导肾透明细胞癌的发生、发展。鉴于NDUFA4L2 作为复合体I 的亚基，在肾透明细胞癌中特异性高表达，而鱼藤酮（Rotenone）作为复合体I 的抑制剂，能够抑制电子信号传递。我们也初步验证了Rotenone能明显抑制肾透明癌细胞的增殖。总之，本研究证实NDUFA4L2作为肾癌特异性肿瘤标志物，在肾癌的诊断、预后中具有重要的价值。同时去初步揭示NDUFA4L2在肾癌发生发展的作用机制以及探讨Rotenone能靶向线粒体复合体I，可能作为肾癌靶向治疗一个潜在药物靶点，从而为肾癌的诊断治疗提供新思路。