Bioinformatics prediction and experimental validation of microRNA-20a targeting Cyclin D1 in hepatocellular carcinoma

Abstract

Hepatocellular carcinoma is the major form of primary liver cancer, which is the second and sixth leading cause of cancer-related death in men and women, respectively. Extensive research indicates that Wnt/β-catenin signaling pathway, which plays a pivotal role in growth, development, and differentiation of hepatocellular carcinoma, is one of the major signaling pathways that is dysregulated in hepatocellular carcinoma. Cyclin D1 is a proto-oncogene and is one of the major regulators of Wnt signaling pathway, and its overexpression has been detected in various types of cancers including hepatocellular carcinoma. Using several validated bioinformatic databases, we predicted that the microRNAs are capable of targeting 3′-untranslated region of Cyclin D1 messenger RNA. According to the results, miR-20a was selected as the highest ranking microRNA targeting Cyclin D1 messenger RNA. Luciferase assay was recruited to confirm bioinformatic prediction results. Cyclin D1 expression was first assessed by quantitative real-time polymerase chain reaction in HepG2 cell line. Afterward, HepG2 cells were transduced by lentiviruses containing miR-20a. Then, the expression of miR-20a and Cyclin D1 was evaluated. The results of luciferase assay demonstrated targeting of 3′-untranslated region of Cyclin D1 messenger RNA by miR-20a. Furthermore, 238-fold decline in Cyclin D1 expression was observed after lentiviral induction of miR-20a in HepG2 cells. The results highlighted a considerable effect of miRNA-20a induction on the down-regulation of Cyclin D1 gene. Our results suggest that miR-20a can be used as a novel candidate for therapeutic purposes and a biomarker for hepatocellular carcinoma diagnosis.

The PI3K/AKT signaling pathway: Associations of miRNAs with dysregulated gene expression in colorectal cancer

“…Overexpression of miR‐17‐5p has been shown to inhibit proliferation and trigger apoptosis . MiR‐20 has previously been associated with cyclin D1 and has been shown to be associated with vascular endothelial growth factor, a cytokine, via activation of Akt signaling . MiR‐27a‐3p has been shown to promote cell proliferation and invasion possibly through its association with the cytokine TRAIL (TNF‐related apoptosis‐inducing ligand) or through its effects on insulin .…”

Retracted: Effect of the Wnt/β‐catenin signaling pathway on apoptosis, migration, and invasion of transplanted hepatocellular carcinoma cells after transcatheter arterial chemoembolization in rats

“…Activation of the Wnt/β‐catenin pathway has been observed in at least 1/3 of HCC, and a significant number of these have mutations in the β‐catenin gene . According to investigation, the Wnt/β‐catenin signaling pathway is involved in the growth, development, differentiation of HCC . A previous study demonstrated that the Wnt/β‐catenin signaling pathway regulates the protein expression of the angiogenic factors, MMP‐2, MMP‐9, VEGF‐A, VEGF‐C, and bFGF in HCC cells .…”

Overexpression of miR‐623 suppresses progression of hepatocellular carcinoma via regulating the PI3K/Akt signaling pathway by targeting XRCC5

“…[26][27][28] It has also been demonstrated that miR-623 targeted cyclin D1 in gastric cancer cells. 7-12 Jiang et al 13 had revealed that miR-623 was downregulated in gastric cancer, and they further found that upregulation of miR-623 inhibited the proliferation in SGC-7901 and BGC-823 cells and enhanced the chemosensitivity of cells to 5-Fluorouracil.…”

“…Cyclin D1 is a key regulator in cell cycle and has been reported to be upregulated in HCC and thought to be a potential target for HCC. [26][27][28] It has also been demonstrated that miR-623 targeted cyclin D1 in gastric cancer cells. 13 In this study, we found that miR-623 could inhibit the cyclin D1 expression in HCC cells.…”