The anticancer effects of cinobufagin on hepatocellular carcinoma Huh‑7�cells are associated with activation of the p73 signaling pathway

Abstract

The Na + /K + -ATPase inhibitor cinobufagin exhibits numerous anticancer effects on hepatocellular carcinoma (HCC) cells expressing wild-type p53 via inhibition of aurora kinase A (AURKA) and activation of p53 signaling. However, the effects of cinobufagin on HCC cells expressing mutant p53 remain unclear. In the present study, the anticancer effects of cinobufagin were investigated on HCC Huh-7 cells with mutant p53, and the effects of AURKA overexpression or inhibition on the anticancer effects of cinobufagin were analyzed. Viability, cell cycle progression and apoptosis of cells were determined using an MTT assay, flow cytometry and Hoechst 33342 staining, respectively. The expression levels of p53 and p73 signaling-associated proteins were investigated via western blot analysis. The results demonstrated that the expression levels of AURKA, B-cell lymphoma 2 (Bcl-2), cyclin-dependent kinase 1, cyclin B1, proliferating cell nuclear antigen and heterogeneous nuclear ribonucleoprotein K, as well as the phosphorylation of p53 and mouse double minute 2 homolog, were significantly decreased in Huh-7 cells treated with 5 µmol/l cinobufagin for 24 h. Conversely, the expression levels of Bcl-2-associated X protein, p21, p53 upregulated modulator of apoptosis and phorbol-12-myristate-13-acetate-induced protein 1, were significantly increased by cinobufagin treatment. Overexpression or inhibition of AURKA suppressed or promoted the anticancer effects of cinobufagin on Huh-7 cells, respectively. These results indicated that cinobufagin may induce anticancer effects on Huh-7 cells via the inhibition of AURKA and p53 signaling, and via the activation of p73 signaling, in an AURKA-dependent manner.

Cinobufagin Suppresses Melanoma Cell Growth by Inhibiting LEF1

“…Although cytotoxic side effects in cardiac myocytes by the administration of a high dose of cinobufagin contained in Chinese Medicine has been reported [37], emerging evidence has shown that cinobufagin effectively suppresses cancer growth and progression by inducing cell cycle arrest and apoptosis and inhibiting angiogenesis [38]. For example, cinobufagin was found to promote cell cycle arrest at the G2/M phase and apoptosis in hepatocellular carcinoma and esophageal squamous cell carcinoma by increasing the expression levels of pro-apoptotic p73 and BAX [39,40]. Cinobufagin was found to induce apoptosis in osteosarcoma cancer cells by suppressing cancer-promoting signaling pathways such as STAT3, Notch and NF-kB, and activating mitochondria-mediated apoptosis pathways [41][42][43][44].…”

WITHDRAWN: A Bioinformatics-Based Screening and Analysis of Key Genes in Hepatocellular Carcinoma

“…And it plays an important role in the mitotic cell cycle by participating in centrosome replication, isolation and maturation [42][43][44]. It was demonstrated that the overexpression or inhibition of AURKA significantly opposed or promoted the anticancer effects of cinobufagin in Huh-7 cells, respectively [45]. Genetic variations in the gene encoding AURKA may be a significant predictor of early HCC occurrence and a reliable biomarker for disease progression [46].…”

Cinobufagin Induces Cell Cycle Arrest at the G2/M Phase and Promotes Apoptosis in Malignant Melanoma Cells

“…Initially, cinobufagin as a painkiller was primitively used to treat pain caused by cancer such as liver, prostate, and breast cancers (16). Recently, it has been reported that cinobufagin can effectively inhibit the proliferation of tumor cells by inducing apoptosis and cell cycle arrest in several tumor cells (e.g., hepatocellular carcinoma Huh-7 cells, colorectal cancer HCT-116 and breast cancer MCF-7 cells) (17–21). Moreover, cinobufagin is one of the chemotherapeutic drugs approved by Chinese State Food and Drug Administration for the treatment of liver and prostate cancers in China (22).…”