Linc-ROR通过活化SP/NK1-R/STAT3信号轴促进胰腺癌神经侵犯及进展的研究

Neural invasion is a common and distinctive biological behavior, which is closely related to the invasion and metastasis, but the mechanism remains unclear. In our previous research, we found that linc-ROR, which is closely related to the neural invasion, promotes the invasion and metastasis of pancreatic cancer. Further preliminary experiments showed that: 1. linc-ROR significantly was overexpressed in pancreatic cancer tissues with neural invasion. 2. High-throughput sequencing suggested that linc-ROR significantly upregulated NK1-R expression. 3. Mechanistically, linc-ROR bound specifically to LOXL2, inhibited its oxidative deamination of chromatin H3K4me3, then transactivated NK1-R transcription, and further activated the JAK2/STAT3 pathway via SP/NK-1R signaling, eventually regulated neural invasion and progression of pancreatic cancer. As mentioned above, we put forward the scientific hypothesis that linc-ROR promotes the neural invasion and progression of pancreatic cancer via SP/NK1-R/STAT3 signal pathway activation. Several important experimental methods including RNA pull-down assay, RIP asssy and sequence truncated mutation test will be used to illuminate the specific molecular mechanism. Based on our previous clinical research of nerve radical dissection of pancreatic cancer, we aim to create a new model based on the combination of nerve radical dissection of pancreatic cancer and targeting the key molecules of neural invasion.

神经侵犯是胰腺癌普遍而独特的生物学行为，与复发转移密切相关，但机制未明。我们前期已报道linc-ROR促进胰腺癌细胞的侵袭转移，且与神经侵犯显著相关。进一步预实验提示：①linc-ROR在神经侵犯组织中高表达；②高通量测序提示linc-ROR显著上调NK1-R表达；③linc-ROR特异性结合LOXL2，遏制其对染色质H3K4me3的氧化去氨基作用，反式激活NK1-R转录，通过SP/NK-1R信号轴激活JAK2/STAT3通路，调控胰腺癌神经侵犯及演进。据此本项目提出“linc-ROR通过活化SP/NK1-R/STAT3信号轴促进胰腺癌神经侵犯及进展”的假说，通过RNA pull-down、RIP和突变序列构建等方法揭示其具体分子机制，结合申请人团队前期在胰腺癌神经清扫根治术中取得的临床研究成果，将为有望开创基于“胰腺癌根治性神经清扫+术后靶向神经侵犯关键调控分子”的组合治疗新模式。

神经侵犯是影响胰腺癌预后的重要因素之一，它具有高发生率，跳跃式转移等特点，与胰腺癌的复发转移密切相关，但其内在机制尚未阐明。本项目基于高通量测序筛选鉴定与胰腺癌神经侵犯高度相关的lncRNA(linc-ROR)，并对其在体内外及临床价值进行验证。主要结果如下：（1）在体外水平上，胰腺癌细胞与神经细胞相互作用的培养模型结果提示linc-ROR通过上调SP/NK1-R信号轴促进胰腺癌细胞的神经侵犯能力；（2）在体内水平上，裸鼠坐骨神经侵犯模型提示linc-ROR促进胰腺癌神经侵犯，而拮抗SP则可抑制该作用；（3）在机制水平上，阐明linc-ROR通过特异性结合LOXL2，遏制其对染色质H3K4me3的氧化去氨基作用，反式激活NK1-R转录，通过SP/NK-1R信号轴激活JAK2/STAT3通路进而促进神经侵犯。（4）临床水平上，qRT-PCR及ISH实验提示linc-ROR在癌灶中高表达，且与神经浸润严重程度呈正相关，生存分析提示linc-ROR高表达的胰腺癌患者预后较差。结合申请人团队前期在胰腺癌神经清扫根治术中取得的临床研究成果，本项目为“胰腺癌根治性神经清扫+术后靶向神经侵犯关键调控分子”的组合治疗新模式提供了新的靶标分子。