Department of Breast and Thyroid Surgery, Provincial Hospital Affiliated To Shandong University, Jingwu Road 324, Huaiyin District, Jinan, 250021, Shandong, China.
Mol Cell Biochem. 2020 Mar;466(1-2):103-115. doi: 10.1007/s11010-020-03692-z. Epub 2020 Jan 31.
Increasing evidence confirmed that the Warburg effect plays an important role involved in the progression of malignant tumors. Resibufogenin (RES) has been proved to have a therapeutic effect in multiple malignant tumors. However, the mechanism of whether RES exerted an antitumor effect on breast cancer through regulating the Warburg effect is largely unknown. The effect of RES on glycolysis was determined by glucose consumption, lactate production, ATP generation, extracellular acidification rate and oxygen consumption rate in breast cancer cells. The total RNA and protein levels were respectively measured by RT-qPCR and western blot. Cell proliferation and apoptosis were examined using the CCK-8 assay, colony formation assay, and flow cytometry, respectively. The interaction between miR-143-3p and HK2 was verified by dual-luciferase reporter gene assay. We also evaluated the influence of RES on the tumor growth and Warburg effect in vivo. RES treatment significantly decreased glycolysis, cell proliferation and induced apoptosis of both MDA-MB-453 and MCF-7 cells. Simultaneously, the expression of HK2 was decreased in breast cancer cells treated with RES, which was positively associated with tumor size and glycolysis. Moreover, HK2 was a direct target gene of miR-143-3p. Mechanistically, upregulation of miR-143-3p by RES treatment inhibited tumor growth by downregulating HK2-mediated Warburg effect in breast cancer. Our findings suggested that RES exerted anti-tumorigenesis and anti-glycolysis activities in breast cancer through upregulating the inhibitory effect of miR-143-3p on HK2 expression, which provided a new potential strategy for breast cancer clinical treatment.
越来越多的证据证实，瓦博格效应在恶性肿瘤的进展中起着重要作用。蟾毒灵（RES）已被证明对多种恶性肿瘤具有治疗作用。然而，RES 是否通过调节瓦博格效应对乳腺癌发挥抗肿瘤作用的机制在很大程度上尚不清楚。通过测定乳腺癌细胞中葡萄糖消耗、乳酸生成、ATP 生成、细胞外酸化率和耗氧量来确定 RES 对糖酵解的影响。分别通过 RT-qPCR 和 Western blot 测定总 RNA 和蛋白质水平。分别用 CCK-8 法、集落形成实验和流式细胞术检测细胞增殖和凋亡。通过双荧光素酶报告基因实验验证 miR-143-3p 和 HK2 之间的相互作用。我们还评估了 RES 对体内肿瘤生长和瓦博格效应的影响。RES 处理显著降低了 MDA-MB-453 和 MCF-7 细胞的糖酵解、细胞增殖并诱导细胞凋亡。同时，RES 处理的乳腺癌细胞中 HK2 的表达降低，与肿瘤大小和糖酵解呈正相关。此外，HK2 是 miR-143-3p 的直接靶基因。从机制上讲，RES 通过下调 HK2 介导的瓦博格效应来上调 miR-143-3p 抑制肿瘤生长。我们的研究结果表明，RES 通过上调 miR-143-3p 对 HK2 表达的抑制作用，在乳腺癌中发挥抗肿瘤和抗糖酵解作用，为乳腺癌的临床治疗提供了新的潜在策略。
