基于Pik3ap1-PI3K/AKT通路探讨围刺促乳腺癌血管正常化作用机制

血管正常化明显提高放化疗及免疫监测点抑制剂疗效。临床与动物实验证实围刺明显提高化疗疗效，但机制不明确。团队前期发现围刺明显促乳腺癌血管正常化，并初步得出正常化时间窗，蛋白组学示围刺通过PI3K/AKT通路促血管正常化，Pik3ap1表达抑制最明显，据此提出假说“围刺通过Pik3ap1-PI3K/AKT通路促血管正常化”。. 本研究拟采用活体光声成像配合组织血管结构功能检测确定围刺促血管正常化时间窗；观察周细胞、内皮细胞超微结构与分子变化，明确血管正常化形态与分子特征。对比观察Pik3ap1-PI3K/AKT通路蛋白变化明确围刺作用靶基因；用CRISPR/Cas9技术建立Pik3ap1敲除4T1乳腺癌模型，观察围刺后Pik3ap1-PI3K/AKT通路蛋白变化验证Pik3ap1是围刺作用核心靶点，阐释围刺促血管正常化分子机制，为肿瘤血管正常化研究提供针刺方案。

Tumor vascular normalization improves the efficacy of radiotherapy, chemotherapy and immune checkpoint inhibitor. Clinical and animal experiments have proved that peritumoral needling can significantly improve the chemotherapy effect, but the mechanism is not clear.Our early study showed that peritumoral needling significantly promoted the local enrichment of chemotherapy drugs in breast cancer, the vascular normalization time window was preliminarily verified by photoacoustic imaging in vivo and detection of SMA, VEGFA and HIF1 expression in tissue. Proteomics indicated that peritumoral needling promoted tumor vascular normalization via PI3K/AKT signaling pathway. Pik3ap1 was a unique member of PI3K family which was significantly inhibited. Hence a hypothesis was put forward that peritumoral needling promoted tumor vascular normalization via Pik3ap1-PI3K/AKT signaling pathway..In this study, we intend to confirm the vascular normalization time window by photoacoustic imaging in vivo and structure and function detection of tumor vasculature in vitro. Changes of ultrastructure and related molecules of pericyte and endothelial cells will be observed to clarify the morphological and molecular characteristics of vascular normalization. The target genes of peritumoral needling will be identified by comparing and observing the protein changes of pik3ap1-pi3k / Akt signaling pathway. The pk3ap1-null 4T1 breast cancer mice model will be established through CRISPR / cas9 technology, and the core role of pik3ap1 will be verified by observing the protein changes of pik3ap1-pi3k / Akt signaling pathway after peritumoral needling treatment. We aim to uncover the molecular mechanism of vascular normalization induced by peritumoral needling, which provides an acupuncture regimen for tumor vascular normalization.