Genistein Restricts the Epithelial Mesenchymal Transformation (EMT) and Stemness of Hepatocellular Carcinoma via Upregulating miR-1275 to Inhibit the EIF5A2/PI3K/Akt Pathway

Abstract

Purpose: Genistein is a natural phytoestrogen with various antitumor effects. In recent years, some microRNAs (miRNA) in cancer cells have been reported to be regulated by genistein. Our study focused on exploring the mechanisms of miRNA upregulation to inhibit the epithelial mesenchymal transformation (EMT) and stemness of hepatocellular carcinoma (HCC). Patients and Methods: MiR-1275 was discovered by the transcriptome sequencing of miRNA expression profiles in HepG2 cells treated with genistein or DMSO as a control. Then, we performed series functional experiments in vitro and vivo to explore the relationship between genistein and miR-1275 in HCC. The target gene (Eukaryotic initiation factor 5A2, EIF5A2) of miR-1275 was predicted by databases and finally determined by a dual luciferase reporter assay. The downstream signaling pathway of EIF5A2 was assessed by bioinformatics analysis and Western blot. Results: the inhibition of genistein on the viability of HCC cells was enhanced by the increase in treatment time and dose, but it had no obvious inhibitory effect on normal hepatocytes (QSG-7701). Through qRT-PCR and transcriptome sequencing, we discovered that miR-1275 was lowly expressed in HCC, and it can be raised by genistein. The overall survival (OS) and recurrence-free survival (RFS) of HCC patients with lowly expressed miR-1275 were lower than those of those with high expression levels. In vitro and vivo experiments exhibited that genistein and the overexpression of miR-1275 can both significantly suppress the proliferation, migration, invasion, metastasis, EMT and stemness of HCC. Moreover, the inhibition can be further enhanced when miR-1275 mimic and genistein exist together. Finally, we demonstrated that miR-1275 can inhibit the epithelial mesenchymal transformation (EMT) and stemness of HCC via inhibiting the EIF5A2/PI3K/Akt pathway. Conclusion: Our findings proved that genistein can inhibit the EIF5A2/PI3K/Akt pathway by upregulating miR-1275 so as to attenuate the EMT and stemness of HCC cells to restrict their progression and metastasis.

Identification of genes that promote PI3K pathway activation and prostate tumour formation

“…Our study shows that chromosome amplifications containing EIF5A2 are associated with various cancers, including prostate, where higher expression levels are seen in amplified tumours. EIF5A2 has been proposed to act in various mechanistic processes, including in the regulation of the PI3K pathway in other tumour types [ 39 – 41 ]. Our data directly connects high levels of EIF5A2 with activation of the PI3K pathway in patient prostate tumours implicating its role in the modulation of this pathway, which has been highly associated with prostate cancer formation and progression.…”

Dietary Epigenetic Modulators: Unravelling the Still-Controversial Benefits of miRNAs in Nutrition and Disease

“…The resveratrol-related modulation of the tumor promoter TGF-β1 was further associated with upregulated miR-663 levels as reported in the SW480 cell line [ 104 ]. Genistein exhibited in vitro and in vivo anticancer effect inhibiting miR-95 and its targets AKT and serum and glucocorticoid-regulated kinase 1 (SGK1) on HCT116 cells [ 105 , 106 ] and enhanced miR-1275 levels thus suppressing Eukaryotic Translation Initiation Factor 5A2 (EIF5A2)/PI3K/AKT pathway in hepatocellular carcinoma [ 107 ]. In mice colorectal cancer tissues, the walnut-based diet PUFAs have been associated with reduced levels of miR-1903, miR-467c, and miR-3068, along with augmented miR-297a expression levels, resulting in anti-inflammatory, antiangiogenetic, antiproliferative and pro-apoptotic effects [ 108 , 109 ].…”

Serum microRNA Profiles and Pathways in Hepatitis B-Associated Hepatocellular Carcinoma: A South African Study

“…Two of these miRNAs in previous HCC studies demonstrated mIR-373-3p targeting TFAP4/CDH1/CSDC2 and PPP6C [72][73][74][75], and miR-1290 targeting SMEK1/FOXC1/SLU7/GLIPR1. The 10 top downregulated miRNAs in terms of LFC ranged between miR-1306-3p (UMI = 2.0; LFC −2.705) modulating FBXL5 [76] and miR-1275 (UMI = 8.0 LFC −2.007) targeting IGFBP1/2/3/E1F5A2 [77][78][79][80][81] and, in general, were less referenced, except for mIR-491-5p (UMI = 3.0; LFC −2.488) targeting PKM2 and SEC61A1/EGFR [82][83][84] and miR-221-5p (UMI = 2.0; LFC −2.146) targeting ITGB5/GCDH and CD44-TGF-B1 [85][86][87][88] (see Supplementary Table S2).…”