Overexpression of Klotho suppresses liver cancer progression and induces cell apoptosis by negatively regulating wnt/β-catenin signaling pathway

Abstract

BackgroundKlotho is a discovered aging suppressor gene, and its overexpression in mice extends the life span of the animal. Recently, Klotho is also identified as a tumor suppressor gene in variety of tumors; however, the potential role and the antitumor mechanism remain unclarified in liver cancers.MethodsRT-PCR and western blotting analysis were used to detect the expression of Klotho, β-catenin, C-myc, and Cyclin D1. MTT assay was used to detect the survival rates of HepG2 and SMMC-7721 cells. Colony formation assay was used to test the proliferation ability in Klotho transfected cells. FACS was used to detect the cell apoptosis rate in different groups.ResultsThe results showed that lower expression of Klotho were found in liver cancer cell lines than the immortalized liver cell L02. Also, MTT assay results found that overexpression or recombinant Klotho administration suppressed the proliferation of liver cancer cells HepG2 and SMMC-7721. Moreover, the colony formation assay results showed that the number of colonies was significantly lower in the cells with transfection with pCMV-Klotho than the controls. Thus, functional analysis demonstrated that Klotho expression inhibited the proliferation of liver cancer cells and Klotho worked as an important antitumor gene in tumor progression. Next, the mechanism was partly clarified that Klotho expression induced cell apoptosis in HepG2 and SMMC-7721 cells, and this phenomenon was mainly involved in the Wnt/β-catenin signaling pathway. The western blotting analysis revealed that overexpression or recombinant administration of Klotho obviously decreased the expression levels of β-catenin, C-myc, and Cyclin D1 in HepG2 cells. Most importantly, the antitumor mechanism for Klotho due to that overexpression of Klotho not only decreased the endogenous β-catenin levels but also inhibited the nuclear translocation of β-catenin to delay the cell cycle progression.ConclusionsKlotho was a tumor suppressor gene, and overexpression of Klotho suppressed the proliferation of liver cancer cells partly due to negative regulation of Wnt/β-catenin signaling pathway. So, Klotho might be used as a potential target, and the study will contribute to treatment for therapy of liver cancer patients.

FLI‐1 mediates tumor suppressor function via Klotho signaling in regulating CRC

“…Our recent study also demonstrated that Toll‐like receptor 4 increased Klotho gene expression, but decreased NF‐κB p65 and IGF‐1R protein phosphorylation (Xie et al, 2019). Klotho functions as a tumor suppressor gene in liver cancer cells through negative regulation of the Wnt/β‐catenin signaling pathway (Sun et al, 2015). In this study, silencing of FLI‐1 gene expression significantly decreased Klotho protein expression and β‐catenin protein phosphorylation, but significantly increased Wnt3a and β‐catenin protein expression and IGF‐1R phosphorylation in HT29 cells; in contrast, overexpressing FLI‐1 gene significantly increased Klotho protein expression and β‐catenin protein phosphorylation, but significantly decreased Wnt3a and β‐catenin protein expression and IGF‐1R phosphorylation in Caco‐2 cells (Figure 5).…”

The suppressive effect of soluble Klotho on fibroblastic growth factor 23 synthesis in UMR‐106 osteoblast‐like cells

“…In a mammalian model of accelerated aging, Klotho suppressed augmented Wnt signaling (Liu et al, ). Overexpression of Klotho or recombinant Klotho administration suppressed the proliferation of liver cancer cells, partly due to negative regulation of the Wnt/β‐catenin signaling pathway (Sun et al, ). In addition, our group confirmed that Klotho inhibited the differentiation of vascular smooth muscle cells into osteoblasts and ameliorated vascular smooth muscle cell calcification partially mediated by the inhibition of the Wnt/β‐catenin signaling pathway (Chen et al, ).…”

Tissue expression and source of circulating αKlotho

“…However, there are some studies indicating that normal liver tissue and cell lines express Klotho mRNA and protein [180,181] and that the levels are decreased in hepatocellular carcinoma (HCC) samples or cell lines [182][183][184][185], and in steatotic liver tissue [186]. Whether low levels of Klotho predict poor outcome in HCC is also a matter of debate [182][183][184][185]187].…”