中药龙葵通过NRP-1基因抑制膀胱移行上皮癌细胞生物学行为

Bladder urothelial carcinoma (BUC) is intractable for high recurrence rate rate. Solasonine (SOL), the biologically active ingredient of traditional Chinese medicine Solanum nigrum, shows anti-tumor activity, but the specific mechanism is unknown. Neuropilin-1 (NRP-1) is a kind of multi function non tyrosine kinase receptor, its high expression is the adverse prognostic factor of BUC. Our previous study determined the BUC cell line models by exploring NRP-1 gene expression abundance in different BUC cells, and found that SOL inhibited T42 BUC cells proliferation in time - and concentration dependentway, accompanied with decreased expression of NRP-1 gene. In the present study, the effects of NRP-1 gene on the biological behavior of BUC cells and its mechanism will be further explored, by expressing lentiviral NRP-1 gene in BUC cells. The regulation of miRNA by traditional Chinese medicine and its active components of has been fully confirmed. Biological information analysis found many miRNA binding sites in 3 'UTR of NRP-1 gene. Thus, we propose the hypothesis that regulation of NRP-1 gene expression by the candidate miRNA gene expression may be one of the main methods of SOL intervention in BUC cells. The experimental verification will be carried out.

膀胱尿路上皮癌（BUC）复发率高，如何治疗是临床难题；中药龙葵的生物活性成分澳洲茄碱（SOL）具有抗肿瘤活性，但是具体机制不明。Neuropilin-1（NRP-1）基因是一类多功能非酪氨酸激酶受体，既往报道其高表达是BUC不良预后因素。我们前期实验通过探寻NRP-1基因在不同BUC细胞中的表达丰度，确定BUC细胞系模型，发现SOL对BUC细胞T42呈时间-浓度依赖性增殖抑制作用，伴随NRP-1基因的表达下降。本实验进一步通过慢病毒干扰NRP-1基因在BUC细胞中的表达，探讨NRP-1基因对BUC细胞生物学行为的影响及其作用机制。中药及其活性成分对miRNA的调节作用已得到充分证实，通过生物信息学分析NRP-1基因3‘UTR存在多个miRNA结合位点，我们提出假说，通过候选miRNA调控NRP-1基因表达可能是SOL干预BUC细胞的主要作用方式之一，并进行实验验证。

通过本课题实验，我们认为龙葵可能通过多种细胞增殖，凋亡逃避，抗炎和血管生成调节机制在膀胱癌患者中起作用。NRP-1慢病毒RNA干扰对膀胱尿路上皮癌细胞的迁移和侵袭具有抑制能力，NRP-1干扰对膀胱尿路上皮癌细胞周围血管新生能力具有抑制能力。且NRP-1在膀胱癌细胞中可以通过MAPK通路，以及通过调控BIRC3、CDK2、CDK4、CDK6、CCNE2、FOS等基因的表达，调节包括细胞增殖，周期，凋亡，侵袭，迁移等生物学行为，为以NRP-1为靶点的抗肿瘤药物和方法提供理论依据。