Oviductus ranae protein hydrolysate (ORPH) inhibits the growth, metastasis and glycolysis of HCC by targeting miR-491-5p/PKM2 axis
Oviductus ranae protein hydrolysate (ORPH) inhibits the growth, metastasis and glycolysis of HCC by targeting miR-491-5p/PKM2 axis
复制标题
哈蟆油蛋白水解物 (ORPH) 通过靶向 miR-491-5p/PKM2 轴抑制 HCC 的生长、转移和糖酵解
DOI：
10.1016/j.biopha.2018.07.071
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发表时间：
2018-11-01
影响因子：
7.5
通讯作者：
Huang, Dongsheng
中科院分区：
文献类型：
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作者：
Xu, Qiuran;Dou, Changwei;Huang, Dongsheng
背景：蛙卵（Oviductus Ranae， OR）是一种珍贵的中药材，具有广泛的生物活性。从OR中提取的蛋白水解物（ORPH）先前被发现具有免疫调节作用和抗胶质瘤活性。本研究旨在探讨ORPH对肝细胞癌（HCC）进展的影响。方法：采用MTT法、BrdU法、菌落形成法和transwell法检测肝癌细胞的体外增殖和移动性。葡萄糖消耗和乳酸生成测定测定HCC细胞的糖酵解。建立裸鼠皮下肿瘤模型和肺转移模型，检测肝癌在体内的生长和转移情况。通过荧光素酶报告基因检测证实miR-491-5p与丙酮酸激酶M2 （PKM2）的3'UTR直接结合。结果：体外实验表明，ORPH能显著抑制HCC细胞的增殖、迁移、侵袭、上皮向间质转化（EMT）和糖酵解。此外，ORPH治疗显著抑制小鼠肝癌的生长和转移。我们证实ORPH能有效降低HCC细胞中PKM2的表达。强制表达PKM2可消除ORPH对HCC细胞的抑制作用。机械上，ORPH通过上调miR-4915p以转录后方式降低PKM2的表达。miR-491-5p在HCC细胞中表现出与ORPH相似的抑瘤作用。此外，ORPH通过调节miR-491-5p/PKM2轴发挥对HCC细胞的抑制作用。最后，miR-491-5p水平降低和PKM2表达升高与HCC患者的不良临床特征和不良预后相关。结论：本研究表明ORPH具有抑制肿瘤生长、转移和糖酵解的作用
Background: Oviductus Ranae (OR) is a valuable Chinese crude drug and has been reported to have a range of biological activities. Protein hydrolysate extracted from OR (ORPH) was previously found to have immune regulatory effect and anti-glioma activity. This study was aimed to investigate the effects of ORPH on hepatocellular carcinoma (HCC) progression.Methods: MTT, BrdU, colony formation and transwell assays were used to determine proliferation and mobility of HCC cells in vitro. Glucose consumption and lactate production assays were carried out to measure the glycolysis of HCC cells. The subcutaneous tumor model and lung metastasis model in nude mice were established to detect tumor growth and metastasis of HCC in vivo. The direct binding of miR-491-5p to 3'UTR of pyruvate kinase M2 (PKM2) was confirmed by luciferase reporter assay.Results: In vitro experiments showed that ORPH significantly inhibited proliferation, migration, invasion, epithelial- to-mesenchymal transition (EMT) and glycolysis of HCC cells. Moreover, ORPH treatment prominently suppressed HCC growth and metastasis in mice. We demonstrated that ORPH effectively decreased the expression of PKM2 in HCC cells. Forced expression of PKM2 abrogated the inhibitory effects of ORPH on HCC cells. Mechanically, ORPH reduced PKM2 expression in a post-transcriptional manner by up-regulating miR-4915p. miR-491-5p exhibited a similar tumor suppressive effects with ORPH in HCC cells. Moreover, ORPH exerted its inhibitory effects on HCC cells through regulating miR-491-5p/PKM2 axis. Lastly, decreased miR-491-5p level and increased PKM2 expression were correlated with unfavorable clinical features and poor prognosis of HCC patients.Conclusions: In all, this study reveals that ORPH inhibits the growth, metastasis and glycolysis