He Jiaming, Wei Qiang, Jiang Rong, Luan Tiankuo, He Shuang, Lu Ruijin, Xu Hang, Ran Jianhua, Li Jing, Chen Dilong
Laboratory of Stem Cells and Tissue Engineering, Department of Histology and Embryology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
Pharmaceuticals (Basel). 2022 Dec 28;16(1):42. doi: 10.3390/ph16010042.
Berberine hydrochloride (BBR) could inhibit the proliferation, migration, and invasion of various cancer cells. As the only enzyme for the de novo synthesis of ribonucleotides, RRM2 is closely related to the development of tumorigenesis. However, not much is currently known about the functional roles of RRM2 in breast cancer (BRCA), and whether BBR regulates the migration and invasion of BRCA cells by regulating the expression of RRM2 remains to be determined. We study the effects of BBR on BRCA cell proliferation in vitro and tumorigenesis in vivo by using colony formation assays, EdU assays, and xenograft models. Transcriptome sequencing, the random forest algorithm, and KEGG analysis were utilized to explore the therapeutic target genes and relative pathways. The expression of RRM2 in BRCA patients was analyzed with The Cancer Genome Atlas (TCGA) dataset, the GEPIA website tool, the Gene Expression Omnibus (GEO) database, and the UALCAN database. The survival probability of BRCA patients could be predicted by survival curve and nomogram analysis. Molecular docking was used to explore the affinity between BBR and potential targets. Gain- and loss-of-function methods were employed to explore the biological process in RRM2 participants. We comprehensively investigated the pharmacological characteristics of BBR on BRCA cell lines and discovered that BBR could inhibit the proliferation of BRCA cells in vitro and in vivo. Combining transcriptome sequencing and KEGG analysis, we found that BBR mainly affected the biological behavior of BRCA cells via HIF-1α and AMPK signal pathways. Additionally, by using bioinformatics and molecular docking, we demonstrated that RRM2 plays an oncogenic role in BRCA samples and that it acts as the hub gene of BBR on BRCA cells. Knockdown and overexpression studies indicated that RRM2 promoted BRCA cell migration as well as invasion in vitro by affecting the epithelial-to-mesenchymal transition (EMT). Our study demonstrated the significance of BBR regulating HIF-1α and AMPK signaling pathways in BRCA cells. Moreover, we revealed the carcinogenic role and potential mechanism of RRM2 as a core regulatory factor of BBR in BRCA in controlling BRCA invasion, migration, and EMT, suggesting that RRM2 may be a therapeutic target and prognostic biomarker for BRCA therapy.
盐酸小檗碱（BBR）可抑制多种癌细胞的增殖、迁移和侵袭。作为核糖核苷酸从头合成的唯一酶，RRM2与肿瘤发生发展密切相关。然而，目前关于RRM2在乳腺癌（BRCA）中的功能作用知之甚少，BBR是否通过调节RRM2的表达来调控BRCA细胞的迁移和侵袭仍有待确定。我们通过集落形成试验、EdU试验和异种移植模型研究了BBR对体外BRCA细胞增殖和体内肿瘤发生的影响。利用转录组测序、随机森林算法和KEGG分析来探索治疗靶点基因和相关途径。使用癌症基因组图谱（TCGA）数据集、GEPIA网站工具、基因表达综合数据库（GEO）和UALCAN数据库分析BRCA患者中RRM2的表达。通过生存曲线和列线图分析预测BRCA患者的生存概率。利用分子对接探索BBR与潜在靶点之间的亲和力。采用功能获得和功能丧失方法探索RRM2参与的生物学过程。我们全面研究了BBR对BRCA细胞系的药理特性，发现BBR可在体外和体内抑制BRCA细胞的增殖。结合转录组测序和KEGG分析，我们发现BBR主要通过HIF-1α和AMPK信号通路影响BRCA细胞的生物学行为。此外，通过生物信息学和分子对接，我们证明RRM2在BRCA样本中发挥致癌作用，并且它是BBR作用于BRCA细胞的枢纽基因。敲低和过表达研究表明，RRM2通过影响上皮-间质转化（EMT）促进体外BRCA细胞的迁移和侵袭。我们的研究证明了BBR调节BRCA细胞中HIF-1α和AMPK信号通路的重要性。此外，我们揭示了RRM2作为BBR在BRCA中的核心调节因子在控制BRCA侵袭、迁移和EMT中的致癌作用及潜在机制，表明RRM2可能是BRCA治疗的治疗靶点和预后生物标志物。
