基于肝素酶-多配体聚糖-外泌体探讨益气活血解毒中药干预三阴性乳腺癌化疗耐药

BACKGROUND: The chemotherapy resistance (CR) of triple negative breast cancer (TNBC) is a big fortress and challenge for doctors and researchers in tumor field. Heparanase (Hpa) has recently been reported to be new target for cancer therapy, as Hpa is expressed at high levels in malignant tumors. It is found that tumor Hpa is higher in CR TNBC patients compared with un-resistance ones by evidence-based medicine research. Although it is well established that Hpa plays a key role in promoting tumor progress,such as enhancing exosome secretion, cuting heparanase sulphate chains of syndecan, promoting metastasis directly. Exosome is one of the main vectors in chemotherapy resistance progress. However, there is no system study on hpa-syndecan-exosome axis while exploring CR mechanism. YIQIHUOXUEJIEDU is one of the successful principles for TCM treatment on tumor, which is widely used for kinds of tumors, and has been proved to be able to reduce tumor proliferation and metastasis abilities, alleviate side-effects of chemotherapy and radiotherapy, increase chemotherapy efficiency. GUBUYILIU II (GY) is the typical prescription based on this principle, which is the key prescription from famous national TCM doctor Ren-Cun Yu, and has been used clinically for more than 20 years. Pilot experiments suggest GYP (modified prescription based on GY) has the potential ability against chemotherapy resistance. By all of these foundings, we come to our hypothesis that hpa-syndecan-exosome axis is one of the key axis of CR in TNBC, higher Hpa level in tumor cut syndecan HS chains leading to its dysfunction, promote exosome formation and secretion, assist chemotherapy resistance; by intervention of hpa-syndecan-exosome axis, GYP can against CR to a certain degree..OBJECTIVE: To investigate the anti-chemoresistance activity of active component alignments isolated from the prescription GYP on triple negative breast cancer models, both in vivo and in vitro (MDA-MB-231-hpa/EPI and MDA-MB-231-mock/EPI); to explore the role of Hpa in chemoresistance; to compare the exosomes from different cells; and to analysis GYP effects on heparanase-syndecan-exosome..EXPECTED RESULTS: The aim of this project is to explore the mechanisms of GYP on triple negative breast cancer chemotherapy resistance, especially by its effects on heparanase, syndecan, and exosome. The expected result of this project is to provide a new therapeutic strategy. Articles published in journals and students educated for master and PhD thesis will be the direct results of this project.

三阴性乳腺癌(TNBC)化疗耐药(CR)是肿瘤治疗难点；CR患者肝素酶(Hpa)水平较高；Hpa剪切多配体聚糖(syndecan)，促进肿瘤转移，促进外泌体(exosome)分泌；exosome是新发现的CR载体。“益气活血解毒法”代表方固本抑瘤II号(GY)是郁仁存教授创制的乳腺癌协定处方，具有较好临床疗效，其变方GYP具有化疗增效及抑制Hpa作用。我们提出假说：高Hpa剪切syndecan影响其功能、促进exosome形成及CR，Hpa-syndecan-exosome轴是TNBC发生CR关键；GYP通过Hpa-syndencan-exosome干预CR。本研究以不同Hpa水平TNBC细胞为工具，从Hpa-syndecan- exosome角度诠释GYP干预化疗耐药作用机理，探讨“益气活血解毒法”干预化疗耐药作用新内涵，为临床更合理地应用该法则治疗肿瘤提供理论指导和实验数据支持。

肿瘤治疗抵抗是限制治疗疗效的重要瓶颈问题，肝素酶高表达是肿瘤不良预后、较短生存时间的独立危险因素。本研究围绕高肝素酶水平肿瘤细胞分泌外泌体介导化疗耐药这一科学问题，以不同肝素酶水平人三阴性乳腺癌细胞（MDA-MB-231-Hpa、MDA-MB-231-mock、MDA-MB-231-WT、MCF-7）及小鼠三阴性乳腺癌细胞4T1，肝素酶转基因鼠（Hpa-tg）及野生型为研究对象，诱导培养不同肝素酶水平乳腺癌表阿霉素耐药株（MDA-MB-231-Hpa/EPI、MDA-MB-231-mock/EPI、MDA-MB-231-EPI），探讨了“益气活血解毒法”代表方固本抑瘤II号、药对及单体的抗肿瘤及干预化疗耐药作用，并从外泌体角度探讨作用机制。实验结果验证修正了研究假说：肝素酶-多配体聚糖-外泌体是肿瘤耐药的关键，肝素酶高表达过度剪切多配体聚糖，促进外泌体释放，化疗耐药；益气活血解毒中药具有一定的抗肿瘤及干预化疗耐药作用，其机制与影响肝素酶表达、外泌体释放有关。研究过程中发表相关论文6篇，指导博士及硕士研究生各2名，衍生获得国家自然基金面上项目1项，研究结果从外泌体层面揭示了中药干预肿瘤作用机制，为临床应用益气活血解毒中药提供实验数据支持。