Chen Xi, Li Qi, Kan Xiao-Xi, Wang Ya-Jie, Li Yu-Jie, Yang Qing, Xiao Hong-Bin, Chen Ying, Weng Xiao-Gang, Cai Wei-Yan, Zhu Xiao-Xin
Capital Medical University School of Traditional Chinese Medicine, Beijing 100069, P.R. China.
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, P.R. China.
Oncol Rep. 2016 Dec;36(6):3215-3224. doi: 10.3892/or.2016.5184. Epub 2016 Oct 19.
Metastasis of breast cancer is the vital step for malignant progression. During such a process, hematogenous metastasis is an indispensable approach for the dissemination of cancer cells. A platelet, contributes to hypercoagulable state, and is also identified the crucial factor in the coagulation system for supporting metastasis. Therefore, the relationship of a platelet and a tumor cell plays a critical role in tumor cell metastasis. Consequently, inhibiting tumor cell‑induced platelet aggregation (TCIPA) is recongnized as a crucial target on suppression of tumor metastasis such as aspirin (ASA). Under such circumstance, here we report that, through dissociating the tumor‑platelet (T‑P) complex, 80% ethanol extracts of Caulis Spatholobi (SET) successfully alleviated the hypercoagulation state, thereby reducing tumor metastasis and improving the prospects of survival in breast cancer cell model. Through MTT and anti‑aggregation assay stimulated by ADP, we detected the optimum treatment time and the optimum dose of SET. By using confocal microscopy, we observed that SET can strongly block the formation of T‑P complex in vitro. The result was further quantified and confirmed by the FACS analysis. The fluorescent value of T‑P complex was obviously decreased in the drug‑treated groups. In vivo, 4T1 cells were injected through the mouse tail vein for dynamic visualization by small animal imaging system. The metastatic intensity was quantified and the survival curve was analyzed. Additionally, general observation and hematoxylin and eosin (H&E) staining of lung tissue was performed. SET exerted an obvious effect on the inhibition of metastasis and increasing the survival rate of mice. For the molecular mechanism study of anti‑TCIPA, zymography and RT‑PCR assay preliminarily revealed the molecular mechanism of SET in the regulation of P‑T interaction. Collectively, through drug efficacy identification and pharmacological revealing, we have obtained a promising candidate for the interference of breast metastasis by suppressing TCIPA, which will be beneficial for clinical cancer treatment.
乳腺癌转移是恶性进展的关键步骤。在此过程中，血行转移是癌细胞扩散不可或缺的途径。血小板有助于形成高凝状态，也是凝血系统中支持转移的关键因素。因此，血小板与肿瘤细胞的关系在肿瘤细胞转移中起着关键作用。因此，抑制肿瘤细胞诱导的血小板聚集（TCIPA）被认为是抑制肿瘤转移的关键靶点，如阿司匹林（ASA）。在这种情况下，我们在此报告，通过解离肿瘤-血小板（T-P）复合物，鸡血藤80%乙醇提取物（SET）成功缓解了高凝状态，从而减少肿瘤转移并改善乳腺癌细胞模型的生存前景。通过MTT和ADP刺激的抗聚集试验，我们检测了SET的最佳治疗时间和最佳剂量。通过共聚焦显微镜观察，我们发现SET在体外能强烈阻断T-P复合物的形成。流式细胞术分析进一步对结果进行了量化和确认。药物处理组中T-P复合物的荧光值明显降低。在体内，通过小鼠尾静脉注射4T1细胞，利用小动物成像系统进行动态可视化。对转移强度进行量化并分析生存曲线。此外，对肺组织进行大体观察和苏木精-伊红（H&E）染色。SET对抑制转移和提高小鼠存活率有明显作用。对于抗TCIPA的分子机制研究，酶谱分析和RT-PCR检测初步揭示了SET调节P-T相互作用的分子机制。总的来说，通过药物疗效鉴定和药理学揭示，我们获得了一种有前景的候选药物，通过抑制TCIPA干扰乳腺癌转移，这将有利于临床癌症治疗。
