中国科学院昆明动物研究所机构知识库

乳腺癌是乳腺上皮组织细胞恶性转化形成的肿瘤，是严重威胁女性身心健康的疾病。在女性肿瘤患者中，乳腺癌的的发病率位居首位，发病率逐年提高，并呈年轻化趋势。乳腺癌患者的死亡率仅次于肺癌。乳腺癌作为一种在分子分型、组织形态和临床表现等有很强异质性的疾病，备受关注。其中，恶性程度高且易复发的三阴性乳腺癌（TNBC）由于缺乏特异的诊断方法、治疗靶点和治疗方法，一直是乳腺癌领域亟待解决的难点问题。TNBC对内分泌治疗和靶向治疗不敏感，所以目前只能按常规化疗治疗。近年来，天然药物成分以其抗肿瘤独特的机制、显著的效果、毒副作用小的特点受到国内外学者广泛的关注。冬凌草（Rabdosia rubescens）是唇形科香茶菜属碎米桠种的草本植物。冬凌草甲素是其主要的有效成分之一。众多研究已经证明冬凌草甲素具有广泛抗炎、抗菌和抗肿瘤作用，包括对乳腺癌的抑制效果也很好。但因冬凌草甲素是萜类化合物，结构复杂、极性差，用药浓度高、生物利用度低等缺点，极大的限制了其作为临床抗肿瘤药物的开发和应用。从药物植物中发掘先导化合物并加以改造合成新型衍生物是开发抗癌药物的新途径。本文以冬凌草甲素为先导化合物，设计合成多个结构新颖未见报道的新型衍生物。通过前期的筛选，发现2’-正丁氨基噻唑并[4,5-a]-1-去氧冬凌草甲素，即CYD-6-28在体外能有效抑制TNBC细胞HCC1806和HCC1937的增殖，抑制作用比冬凌草甲素分别提高约17倍、6倍，具有明显的抗癌活性。进一步研究发现，CYD-6-28能够显著抑制TNBC的细胞DNA合成，并将细胞周期阻滞在G2/M期。更重要的是，CYD-6-28能明显诱导TNBC细胞发生凋亡。实验证明CYD-6-28能显著上调p21表达，激活Caspase-3、-7、-8及其底物PARP 的切割，并下调Cyclin D1、FLIPL、XIAP等蛋白的表达水平。CYD-6-28还能抑制细胞信号转导分子STAT3 、AKT 的激活，增强ERK的激活。CYD-6-28诱导TNBC细胞凋亡的部分是通过诱导DR5的上调表达而实现的。体内实验证明，CYD-6-28能有效抑制荷瘤裸鼠乳腺癌细胞的生长，对实验动物的毒副作用小，用药安全性较高。 本项研究首次报道了冬凌草甲素新型衍生物CYD-6-28通过调控与细胞周期、凋亡相关蛋白的表达，明显抑制TNBC细胞的生长，诱导TNBC细胞凋亡。具有开发为治疗TNBC药物的潜力。CYD-6-28的结构新颖，极性提高，抗肿瘤作用优于冬凌草甲素。目前未见国内外的相似文献报道。该研究对进一步开发利用冬凌草甲素，开发植物来源的抗肿瘤药物，推动中医药现代化有着重要的科学意义。

Breast cancer is a malignant tumor which occurs in the breast epithelial tissue. It remains the leading cause of death among women with cancer in the world. Triple negative breast cancer ( TNBC ) is a distinct subtype of breast cancer with unique pathologic, molecular and clinical behavior. It is a basal-like tumor associated with high malignancy, high recurrence rate and poor prognosis. It poses a therapeutic challenge due to the lack of the conventional targeted drugs and targeted therapies. Currently, chemotherapy drugs are commonly used to treat TNBC, although it is effective, but the patient is suffering from pain and easy to relapse. In recent years, compounds from natural herbs has been widely concerned by scholars for its unique mechanism, remarkable effect and little side effect. Oridonin is a natural anti-cancer compound that was originally isolated from the traditional Chinese herb Rabdosia rubescens. It has extensive pharmacological and physiological effects, including anti-inflammatory, antibacterial and antitumor effects, including breast cancer. However, the clinical application of oridonin are far from ideal due to inferior aqueous solubility and poor cell permeability. It is necessary to synthesize the novel anti-tumor compounds for the clinical treatment. In this study, we investigated a series of newly designed oridonin analogs in terms of their actions against HCC1806 and HCC1937 TNBC cell lines and found that CYD-6-28 significantly inhibits cancer cell proliferation and induces G2/M phase arrest and apoptosis. CYD-6-28 induces the expression of p21 and the cleavage of Caspase-3, -7, -8 and PARP and inhibits the expression levels of Cyclin D1, FLIPL and XIAP. CYD-6-28 also inhibits the activations of STAT3 and AKT and induces the activation of ERK. We demonstrated that CYD-6-28 induces apoptosis at least partially by inducing the expression of death receptor 5 (DR5). Finally, CYD-6-28 significantly suppresses HCC1806 xenograft tumor growth in nude mice at 5 mg/kg without significant toxicity. Taken together,it is the first to report the anti-tumor in TNBC. Itindicates that CYD-6-28 has the potential to be developed as a therapeutic agent to treat TNBC. The analog has novel structure and superior anti-tumor effect. The research will promote the further development and utilization of oridonin and the modernization of Traditional Chinese medicine.