Huang Di, Huang Yu, Huang Zisheng, Weng Jiefeng, Zhang Shuai, Gu Weili
Department of Surgery, Guangzhou First People's Hospital, School of Medicine, South China University of Technology, No.1 Panfu Road, Yuexiu District, Guangzhou, 510180 Guangdong China.
Cancer Cell Int. 2019 Jun 18;19:166. doi: 10.1186/s12935-019-0885-z. eCollection 2019.
New therapeutic drug for breast cancer (BRCA), especially triple negative BRCA (TNBC), is urgently needed. Even though 2-(4-morpholinoanilino)-6-cyclohexylaminopurine (reversine) is an aurora kinase inhibitor, it also inhibits some cancer cells and human BRCA cells. However, the potential roles of reversine as a novel therapeutic agent for the treatment of BRCA remains unknown and must be further investigation. Thus, the relationship of reversine to aurora kinase in BCRA has not been reported. The relationship between AURKB and survival rate in BRCA has never been reported. Herein, we tested the roles of reversine on different BRCA cell line subtypes. We also investigated the relationship between AURKB and survival rate in BRCA as well as reversine to Aurora kinase expression in BCRA cell lines, including TNBC subtype, 4T1, MDA-MB-231, and luminal subtype MCF-7.
Cell viability and apoptosis were detected using Cell Counting Kit-8 and flow cytometry analysis, respectively. Apoptotic and tumor-related proteins were tested using Western blot analysis. Important microRNAs that regulate BRCA were analyzed using RT-PCR. UALCAN public databases were used to analyze the targeted gene profiles, and the PROGgeneV2 database was used to study the prognostic implications of genes.
Reversine inhibits cell proliferation and induces cell apoptosis by modulating caspase-3 and bax/bcl-2 among the three cell lines. Data from the UALCAN public database show that BRCA tissues expressed high gene levels of , and compared with the normal tissue. Among the over-expressed genes in BRCA, ranks 9th in TNBC, 49th in luminal subtype, and 48th in HER2 subtype. High level in BRCA is highly related to the low survival rate in patients displayed in 18 databases searched via PROGgeneV2. The protein levels of aurora B kinase (Aurora B), which is encoded by gene, are highly suppressed by reversine in the three cell lines. The tumor-related proteins TGF-β1, TIMP1, and MMP9 are partially suppressed by reversine but with different sensitivity in the three cell lines. The reversine-affected microRNAs, such as miR129-5p, miR-199a-3p, and miR-3960, in MDA-MB-231 cell line might be the research targets in TNBC regulation.
In BRCA, the level of are over-expressed and is related to low survival rate. Reversine contributes to anti-growth effect in BRCA cell lines, especially for TNBC, by modulating the aurora B. However, the invasiveness, metastasis, and anti-tumor effects of reversine in vivo and in vitro must be further investigated.
乳腺癌（BRCA），尤其是三阴性乳腺癌（TNBC），急需新的治疗药物。尽管2-（4-吗啉代苯胺基）-6-环己基氨基嘌呤（reversine）是一种极光激酶抑制剂，但它也能抑制一些癌细胞和人BRCA细胞。然而，reversine作为一种新型治疗药物治疗BRCA的潜在作用仍不清楚，必须进一步研究。因此，尚未有关于reversine与BRCA中极光激酶关系的报道。AURKB与BRCA生存率之间的关系也从未被报道过。在此，我们测试了reversine对不同BRCA细胞系亚型的作用。我们还研究了AURKB与BRCA生存率之间的关系，以及reversine与BRCA细胞系（包括TNBC亚型、4T1、MDA-MB-231和管腔亚型MCF-7）中极光激酶表达的关系。
分别使用细胞计数试剂盒-8和流式细胞术分析检测细胞活力和凋亡。使用蛋白质免疫印迹分析检测凋亡和肿瘤相关蛋白。使用逆转录-聚合酶链反应分析调控BRCA的重要微小RNA。使用UALCAN公共数据库分析靶向基因谱，并使用PROGgeneV2数据库研究基因的预后意义。
reversine通过调节三种细胞系中的半胱天冬酶-3和bax/bcl-2来抑制细胞增殖并诱导细胞凋亡。UALCAN公共数据库的数据显示，与正常组织相比，BRCA组织中 、 和 的基因水平较高。在BRCA中过表达的基因中， 在TNBC中排名第9，在管腔亚型中排名第49，在HER2亚型中排名第48。通过PROGgeneV2搜索的18个数据库显示，BRCA中高水平的 与患者低生存率高度相关。由 基因编码的极光B激酶（Aurora B）的蛋白水平在三种细胞系中被reversine高度抑制。肿瘤相关蛋白TGF-β1、TIMP1和MMP9被reversine部分抑制，但在三种细胞系中的敏感性不同。在MDA-MB-231细胞系中，受reversine影响的微小RNA，如miR129-5p、miR-199a-3p和miR-3960，可能是TNBC调控中的研究靶点。
在BRCA中， 的水平过表达且与低生存率相关。reversine通过调节极光B对BRCA细胞系，尤其是TNBC，发挥抗生长作用。然而，reversine在体内和体外的侵袭性、转移性和抗肿瘤作用必须进一步研究。
