从肝素酶-硫酸肝素-自噬探讨益气活血解毒中药干预乳腺癌转移的机制

BACKGROUND: YIQIHUOXUEJIEDU is one of the successful principles for TCM treatment on tumor, which is widely used for kinds of tumor clinic treatment, and has been proved to can reduce tumor proliferation and metastasis, alleviate side-effects of chemotherapy/radiotherapy. GUBUYILIU II (GY) is the typical prescription based on this principle. Heparanase and heparan sulfate have recently been reported to be new targets for cancer therapy, as heparanase often is expressed at high levels in malignant tumors. Autophagy, which is constitutively executed at basal level in all cells, promotes cellular homeostasis by regulating organelles and proteins turnover. In tumor cells, autophagy is activated in response to various cellular stresses, including nutrient and growth factor starvation, as well as hypoxia. It is now well established that autophagy can act as tumor suppressor and tumor promoter. The different roles of autophagy in cancer cells seem to depend on tumor type, stage, and genetic context. Recently it is reported that heparanase enhances tumor growth and chemoresistance by promoting autophagy. Since GY is found to reduce tumor proliferation and metastasis by inhibit heparanase expression, and autophagy is one of the target of GY on breast cancer, we come to our hypothesis that by heparanase sulfate structure modification, heparanase promote autophagy, and GY reduce tumor proliferation and metastasis by heparanase, heparan sulfate, and autophagy.. OBJECTIVE: To investigate the anti-tumor activity of active component alignments isolated from the prescription GY on both mice models and breast cancer cells models (heparanase transgenic mice/cells and wide type mice/cells), to investigate its effects on heparanase, heparan sulfate, and autophagy in vitro and in vivo.. EXPECTED RESULTS: The aim of this project is to make clear of GY mechanisms on breast cancer metastasis, expecially by its effects on heparanase, heparan sulfate, and autophagy. The expected result of this project is to provide a new therapeutic strategy. Articles published in journals and patents will be the direct results of this project.

虚、瘀、毒是乳腺癌的病机要点，基于益气活血解毒法的固本抑瘤II号(GY)是肿瘤专家郁仁存教授多年经验总结创制的代表方药，具有良好的临床疗效。肝素酶(HPSE)切割底物硫酸肝素(HS)，影响其结构和功能，进而影响多种下游信号转导。基于高HPSE水平的微环境中自噬(AP)增多、细胞死亡减少而促进肿瘤转移的最新研究发现，分析我们前期证实的GY对AP的双向调节作用、对HPSE的抑制作用，我们提出假说：作为HPSE的唯一底物，HS结构的变化是开启自噬-存活/自噬-死亡的关键；GY通过干预HS结构，影响HPSE及AP信号转导，抑制乳腺癌转移。本研究以不同HPSE水平小鼠/细胞为研究工具，从肝素酶-硫酸肝素-自噬角度探讨“益气活血解毒”法单独应用及与西药联合应用的作用及机理，诠释其抗肿瘤作用的新内涵，为临床更好应用该中药提供理论指导和实验数据支持。

益气活血解毒是针对肿瘤“虚、瘀、毒”证候要点的重要治疗法则，益气活血解毒代表方剂固本抑瘤II号(GY)是郁仁存多年经验总结的该法则有效代表方药。肝素酶(HPSE)切割底物硫酸肝素(HS)，影响其结构和功能，进而影响多种下游信号转导。基于高HPSE水平的微环境中AP增多 、细胞死亡减少而促进肿瘤转移的最新研究发现，分析我们前期证实的GY对AP的双向调节作用、对HPSE的抑制作用，我们提出假说:作为HPSE的唯一底物，HS结构的变化是开启自噬-存活/自噬-死亡的关键;GY通过干预HS结构，影响HPSE及AP信号转导，抑制乳腺癌转移。通过以益气活血解毒中药为工具，以不同肝素酶表达水平的乳腺癌细胞株、以及相应的乳腺癌乳垫原位移植模型作为研究对象，观察益气活血解毒中药对不同肝素酶水平的乳腺癌细胞生长、迁移能力的影响；益气活血解毒中药（与自噬抑制剂联用）对乳腺癌细胞自噬过程的影响及相关信号通路的调控；对肝素酶高表达小鼠乳腺癌肿瘤生长过程中血管生成及自噬的干预作用；以及益气活血解毒中药的体内安全性评价等，我们证实了肝素酶对肿瘤生长、转移的促进作用，本研究中所采用的益气活血解毒解毒中药诱导了肿瘤细胞自噬过程，引起自噬性死亡，但此效应与肝素酶无直接关联。益气活血解毒中药可通过上调自噬过程等相关机制抑制肿瘤发展，具有良好的研究前景。