活血化瘀治法药组靶向FBP1激活HIF-1α/PDK1轴调控糖代谢重编辑干预Her-2阳性乳腺癌的作用及机制研究

乳腺癌发病率高且易复发和转移。活血化瘀是中医临床治疗乳腺癌的常用治法，有报道称其对Her-2阳性乳腺癌更适宜，但未见相关的基础研究。本课题组发现：活血化瘀治法药组（PBC）能抑制DMBA诱导型乳腺癌，RNA-SEQ筛选出糖异生限速酶FBP1，RT-PCR证实PBC使FBP1mRNA升高、乳酸含量与LDHA、PDK1mRNA降低。FBP1不仅抑制糖酵解，且能抑制核中HIF-1的功能，下调HIF-1下游的糖酵解与氧化磷酸化交联的守门基因PDK1，进行糖代谢重编辑。鉴于此提出假说：PBC靶向升高FBP1，激活HIF-1α/PDK1轴，调控糖代谢重编辑，促进线粒体细胞凋亡、抑制Her-2乳腺癌增殖。该研究将采用UPLC-MS/MS、CRISPR／Cas9敲除等技术，结果将明确活血化瘀治法药组对Her-2乳腺癌的作用与机制，丰富与发展活血化瘀治法的临床前研究，推进中医治法在乳腺癌上的精准应用。

Breast cancer has a high incidence and is prone to recurrence and metastasis. The therapy of promoting blood circulation (PBC) (Huo Xue Hua Yu Therapy) is a common treatment for clinical treatment of breast cancer in Traditional Chinese Medicine. Reports showed this therapy had better efficacy for Her-2 positive breast cancer, but no related basic research to support this version. Our research team found that the PBC can inhibit DMBA-induced breast cancer, and found out the gluconeogenesis rate limiting enzyme FBP1 by RNA-SEQ . We confirmed that PBC could increase FBP1 mRNA, decrease LDHA and PDK1 mRNA by RT-PCR, also decrease lactic acid content. FBP1 not only directly inhibits the glycolysis process, but also inhibits the function of HIF-1 in the nucleus, which will down-regulate its target gene PDK-1,also a glycolytic rate-limiting enzyme, which have an obviously effect on reprogramming glucose metabolism on cancer. So as that a hypothesis is proposed: PBC will target to elevate FBP1 and regulate the HIF-1α/PDK1 axis, inhibit glycolysis and promote oxidative phosphorylation in order to reprogram glucose metabolism. PBC promote apoptosis of mitochondria and inhibit the proliferation of Her-2 breast cancer. In this study, we will use UPLC-MS/MS and CRISPR/cas9 knock-out techniques. The results will clarify the effect and mechanism of the Huo Xue Hua Yu Therapy on Her-2 breast cancer, enrich and develop preclinical studies of Huo Xue Hua Yu Therapy, and promote the accurate application of TCM treatment in breast cancer.