RBMX靶向hnRNPA1蛋白调控PKM可变剪接抑制膀胱癌发生发展的功能机制研究

膀胱癌是我国最常见的泌尿系统恶性肿瘤之一，缺乏有效的治疗方法，抑制肿瘤细胞能量代谢是潜在的靶向治疗途径。我们前期采用生物信息学分析筛选及临床标本验证，发现RBMX与膀胱癌恶性进展呈负相关；体外细胞实验证实RBMX显著抑制膀胱癌细胞增殖、克隆形成、迁移和侵袭；CO-IP和MS技术初步确定RBMX的相互作用蛋白是hnRNPA1。PKM是肿瘤能量代谢的关键酶且受hnRNPA1剪接调节。因此，推测RBMX靶向hnRNPA1蛋白调控PKM可变剪接抑制膀胱癌的发生发展。本项目拟在小鼠体内研究RBMX对膀胱癌细胞成瘤及肺转移的能力，体外细胞实验确定RBMX-hnRNPA1-PKM的靶向结合作用，阐明RBMX靶向hnRNPA1蛋白调控PKM可变剪接抑制膀胱癌发生发展的机制，最后，临床标本和随访资料研究RBMX与膀胱癌临床分期、进展和预后的关联性。本项目顺利完成有望为膀胱癌的诊断和靶向治疗提供新科学依据。

Bladder cancer is one of the most common malignant tumors of urinary system in China, but there is a lack of effective treatment. Inhibition of energy metabolism of tumor cells is a potential targeted treatment. Our bioinformatics and clinical tissue samples results showed that RBMX was negatively correlated with the malignant progression of bladder cancer. The in vitro cell experiments confirmed that RBMX significantly inhibited the proliferation, clonal formation, migration and invasion of bladder cancer cells. CO-IP and MS technology preliminarily confirmed that the interaction protein of RBMX is hnRNPA1. PKM, a key enzyme of tumor energy metabolism, is regulated by hnRNPA1 splicing. Therefore, it is speculated that RBMX targeting hnRNPA1 regulates PKM alternative splicing to inhibit the occurrence and development of bladder cancer. The purpose of this project is to study the effect of RBMX on tumorigenesis and pulmonary metastasis of bladder cancer cells in mice, to determine the targeted binding relationship of RBMX-hnRNPA1-PKM in vitro cell experiments, and to elucidate the molecular mechanism of RBMX targeting hnRNPA1 protein to regulate PKM alternative splicing to inhibit the occurrence and development of bladder cancer. Finally, clinical specimens and follow-up data were used to study the relationship between RBMX and clinical stage, disease progression and prognosis of bladder cancer. The successful completion of this project is expected to provide a new scientific basis for diagnosis and targeted treatment of bladder cancer.