环状RNA circ_3763调控GSK3β诱导胰腺癌细胞EMT及转移的机制研究

Pancreatic cancer has a dismal prognosis due to high incidence of metastases. Microarray expression profile of circular RNAs in human pancreatic cancer and bioinformatics analysis showed that a large number of circRNA participation in development and incidence of pancreatic cancer. Further find Hsa_circ_0003763 and glycogen synthase kinase-3 beta (GSK3beta) in pancreatic cancer tissue and metastases tissue are both high expression, and its high expression was significantly associated with lymph node metastasis. Biological information predict GSK3beta is Hsa_circ_0003763 target genes. Down-regulated GSK3β inhibit pancreatic cancer cell proliferation and invasive ability. So we could speculated that Hsa_circ_0003763 regulating GSK3beta promotes epithelial-to-mesenchymal transition and metastases in pancreatic cancer cells. We intends to further research the relationship between Hsa_circ_0003763-miRNAs-GSK3beta and the invasion and metastasis in pancreatic cancer by using of luciferase report carrier, in situ hybridization, RNA-Sequencing detection, Matrigel invasion experiment and PCR site-directed mutation. This topic research will reveal the molecular mechanism that Hsa_circ_0003763 regulate GSK3beta signaling pathways by inhibits micro RNA leading to epithelial-to-mesenchymal transition and metastasis in pancreatic cancer and will provide new targets for the cancer therapy.

转移是影响胰腺癌患者预后的重要因素。本课题前期利用circRNA基因芯检测6例胰腺癌及癌旁组织，生物信息学分析表明大量的circRNA参与胰腺癌的发生发展。进一步发现circ_3763和GSK3β在胰腺癌都高表达，且其表达增强与淋巴结转移显著相关。生物学信息预测GSK3β是circ_3763的靶基因，下调GSK3β能抑制胰腺癌细胞的侵袭及转移能力。因此我们推测circ_3763可能调控GSK3β促进胰腺癌细胞EMT及转移。我们拟进一步构建荧光素酶报告载体、原位杂交、RNA-Seq检测、Matrigel侵袭实验、PCR定点突变等方法在动物成瘤、组织、细胞和分子水平进一步阐明circ_3763、miRNA、GSK3β网络调控机制；本课题研究将揭示circ_3763通过miRNA调节GSK3β信号通路促进胰腺癌EMT及转移的作用及分子机制，为肿瘤的治疗提供新靶点。

本项目原计划研究circRNA3763通过调节GSK3β信号影响胰腺癌EMT及转移的作用及分子机制，为肿瘤的治疗提供新的理论依据。在获得国家自然科学青年基金资助后，该项目进展顺利。本研究观察到胰腺癌组织中circRNA3763高表达以及GSK3β低表达与肿瘤病例分期，预后呈负性相关，即：高表达circRNA3763患者预后更差，低表达GSK3β患者预后相对更差。体外研究显示沉默circRNA3763后，PANC-1细胞增殖明显受到抑制，细胞迁移能力减弱，EMT指标降低，而过表达GSK3β可部分逆转上述过程。PANC-1细胞过表达circRNA3763，引起miR148a、miR26a-5p下调，促进panc-1细胞增殖，迁移，抑制肿瘤细胞凋亡。动物试验显示circRNA3763过表达可加剧胰腺癌肿瘤转移。