Tiliroside as a CAXII inhibitor suppresses liver cancer development and modulates E2Fs/Caspase-3 axis

Rui Han,Hongxing Yang,Lingeng Lu,Lizhu Lin

doi:10.1038/s41598-021-88133-7

Abstract

Liver cancer is the fatal cause of cancer deaths worldwide due to its aggressiveness and lack of effective therapies. Tiliroside (C30H26O13) is an active compound extracted from herb plant Tribulus terrestris L., which has been used as alternative therapy in clinic practice. However, its therapeutic use against liver cancer has not been previously reported. Here, we showed that Tiliroside exerted significantly higher anti-proliferation effect on liver cancer cell lines Hep3B and SNU-449 than on liver normal cell THLE-3 cells or NC group, respectively, by using MTS assay. Results from colony formation, immigration and invasion assays support the anticancer efficacy of Tiliroside and its low-toxic property while treating liver normal cell THLE-3. 3D spheroid formation and CD133 expression level also displays its anti-stemness effect. It has been showed that Tiliroside may function as Carbonic anhydrases XII (CAXII) inhibitor and affects apoptotic E2F1/E2F3/Caspase-3 axis by using CAXII esterase activity assay, Human carbonic anhydrase 12 (CA-12) ELISA Kit, quantitative reverse transcription PCR (RT-qPCR) as well as CaspACE Assay System, respectively. In summary, we demonstrate for the first time that Tiliroside suppresses liver cancer development possibly by acting as a novel CAXII inhibitor, which warrant further investigation on its therapeutic implications.

Highlights

Hepatocellular carcinoma (HCC) is the most frequent pathological type of liver cancer, which ranks the top six in the most common malignant tumors and cancer-caused death worldwide[1]
The inhibition rates of Hep3B in the presence of 40 μM Tiliroside ranged from 27.15 ± 3.7% to 52.27 ± 3.61% during 48 h to 96 h, and displayed significantly higher inhibition rates when compared to the 20 μM Tiliroside group with the range from 6.88 ± 0.8% to 11.57 ± 1.03% (P < 0.001, from 24 to 96 h) (Fig. 1A)
The inhibition rate of SNU-449 in the Tiliroside group ranged from 35.39 ± 5.82 to 55.74 ± 1.85, displayed significant differences to 20 μM Tiliroside group from 24 to 96 h (P < 0.001, from 24 to 96 h, respectively) (Fig. 1B)

Summary

Introduction

Hepatocellular carcinoma (HCC) is the most frequent pathological type of liver cancer, which ranks the top six in the most common malignant tumors and cancer-caused death worldwide[1]. The modulation of NF-κB signaling in liver cancer cells is one of the molecular mechanisms underlying TT extracts-induced apoptosis and proliferation suppression[5]. E2F1 and E2F3, on the other hand, are transcription factors that are involved in the maintenance and self-renewal of cancer stem cells (CSCs), and cancer cell apoptosis by regulating Caspase-3 activity which is a crucial apoptosis e xecutor[18,19,20,21]. Overexpression of these genes has been shown to increase mortality risk in human cancer including breast, ovarian and liver c ancer[18,19,22]. Functional experiments were carried out to confirm the CAXIIinhibiting role of Tiliroside and its impact on apoptosis-related pathways (Supplementary Information)

Objectives

Methods

Results

Conclusion