MiR-448 suppresses cell proliferation and glycolysis of hepatocellular carcinoma through inhibiting IGF-1R expression

Yilang Wang; Xiaohong Chen; Ninghua Yao; Jun Gong; Yongfeng Cao; Xiaoqing Su; Xiu Feng; Min Tao

doi:10.21037/jgo-22-90

MiR-448 suppresses cell proliferation and glycolysis of hepatocellular carcinoma through inhibiting IGF-1R expression

J Gastrointest Oncol. 2022 Feb;13(1):355-367. doi: 10.21037/jgo-22-90.

Authors

Yilang Wang^#^{1

2}, Xiaohong Chen^#³, Ninghua Yao^#⁴, Jun Gong², Yongfeng Cao², Xiaoqing Su², Xiu Feng⁴, Min Tao^{1

5}

Affiliations

¹ Department of Oncology, The First Affiliated Hospital of Soochow University, Suzhou, China.
² Department of Medical Oncology, The Affiliated Tumor Hospital of Nantong University, Nantong, China.
³ Department of Ultrasound Medicine, The Second Affiliated Hospital of Nantong University, Nantong, China.
⁴ Department of Oncology, Affiliated Hospital of Nantong University, Nantong, China.
⁵ Department of Oncology, Dushu Lake Hospital Affiliated to Soochow University, Suzhou, China.

^# Contributed equally.

Abstract

Background: Microribonucleic acids (miRNAs) have been shown to play important roles in hepatocellular carcinoma (HCC) progression. MiR-448 has frequently been shown to be a tumor suppressor, and is abnormally expressed in HCC tumor tissues. However, little is known about the role of miR-448 in HCC development. In this article, the regulatory role of miR-448 on insulin-like growth factor 1 receptor (IGF-1R) in modulating hepatoma cell viability and glycolysis was investigated.

Methods: The expression of miR-448 profiles in clinical tumor tissues and cell lines was examined using quantitative real-time polymerase chain reaction (qRT-PCR). HepG2 and Huh7 cells were transfected with miR-448 mimics, inhibitors, and scramble sequences. Cell viability and apoptosis were determined by a Cell Counting Kit-8 assay and a flow cytometry analysis. IGF-1R, a potential target of miR-448, was selected following a bioinformatic analysis, and the regulatory effects of miR-448 on IGF-1R expression was confirmed by luciferase reporter assay, qRT-PCR, and western blot. Glucose uptake, lactate production, and adenosine triphosphate (ATP) generation were detected by corresponding kits.

Results: Decreased miR-448 expression was observed in both HCC patients' tumor tissues and hepatoma cells in vitro. The overexpression of miR-448 in HepG2 and Huh7 cells decreased cell viability and increased apoptosis. Additionally, the overexpression of miR-448 or the knockdown of IGF-1R lowered the level of glucose uptake, lactate production, and ATP generation, while the knockdown of miR-448 increased glycolysis. Further, aberrantly expressed miR-448 downregulated IGF-1R levels, while the inhibition of miR-448 resulted in the upregulation of IGF-1R in both HepG2 and Huh7 cells. In addition, miR-448 interacted with the wild-type 3'untranslated regions (3'UTRs) of IGF-1R, but had no effect on the mutant 3'UTRs. The expression of IGF-1R was increased in HCC patients' tumor tissues and serum, and was inversely correlated with miR-448 expression.

Conclusions: The increased expression of miR-448 appears to downregulate the expression of IGF-1R by interacting with the 3'UTR in HCC progression. These findings highlight its role as a potential target for HCC therapy.

Keywords: Hepatocellular carcinoma (HCC); apoptosis; glycolysis; insulin-like growth factor 1 receptor (IGF-1R); miR-448.