N-Acetyltransferase 10 Enhances Doxorubicin Resistance in Human Hepatocellular Carcinoma Cell Lines by Promoting the Epithelial-to-Mesenchymal Transition.

Xiuming Zhang,Shi Jiang,Jiang Chen,Rui Ma,Shilin He,Yanfeng Bai,Linghua Zhu,Xiao Liang

doi:10.1155/2019/7561879

Abstract

Background N-Acetyltransferase 10 (NAT10) has been reported to be expressed at high levels in hepatocellular carcinoma (HCC); however, its role in chemoresistance is unclear. This study is aimed at investigating whether NAT10 regulates the epithelial-mesenchymal transition (EMT) and chemoresistance in HCC. Methods HCC cell lines (Huh-7, Bel-7402, SNU387, and SNU449) were treated with remodelin, an inhibitor of NAT10, or transfected with small inhibitory RNAs (siRNAs) targeting NAT10 or Twist. The EMT was induced by hypoxia. The CCK-8 assay was used to quantify cell viability, the EdU incorporation assay to assess cell proliferation. siRNA knockdown efficiency and epithelial/mesenchymal marker expression were assessed by western blotting. Results Knockdown of NAT10 using siRNA or inhibition of NAT10 using remodelin increased the sensitivity of HCC cell lines to doxorubicin; similar effects were observed in cells transfected with the Twist siRNA. Inhibition of NAT10 using remodelin also reversed the ability of doxorubicin to induce the EMT in HCC cells. Furthermore, inhibiting NAT10 reversed the hypoxia-induced EMT. Finally, we confirmed that combining doxorubicin with remodelin delayed tumor growth and reduced tumor cell proliferation in a mouse xenograft model of HCC. Conclusions NAT10 may contribute to chemoresistance in HCC by regulating the EMT. The mechanism by which NAT10 regulates the EMT and doxorubicin sensitivity in HCC cells merits further investigation.

Highlights

Hepatocellular carcinoma (HCC) is the sixth most common malignant cancer worldwide
We examined the cell viabilities of hepatocellular carcinoma (HCC) cells treated with doxorubicin and remodelin, an inhibitor of N-Acetyltransferase 10 (NAT10), for 48 h
The Ethynyl Deoxyuridine (EdU) incorporation assay confirmed that the inhibition of NAT10 using remodelin decreased the proliferation of all four HCC cell lines when treated with doxorubicin (Figures 1(e)–1(h) and Table 1)

Summary

Introduction

Hepatocellular carcinoma (HCC) is the sixth most common malignant cancer worldwide. The 5-year overall survival rate for HCC is very low [1, 2], and the poor prognosis is mainly attributed to acquisition of chemoresistance during therapy [3]. The EMT is implicated in the progression of cancer, and in recent decades, the EMT has been confirmed to play a role in the chemoresistance of various carcinomas, including HCC [11, 12]. N-Acetyltransferase 10 (NAT10) has been reported to be expressed at high levels in hepatocellular carcinoma (HCC); its role in chemoresistance is unclear. This study is aimed at investigating whether NAT10 regulates the epithelialmesenchymal transition (EMT) and chemoresistance in HCC. Knockdown of NAT10 using siRNA or inhibition of NAT10 using remodelin increased the sensitivity of HCC cell lines to doxorubicin; similar effects were observed in cells transfected with the Twist siRNA. Inhibition of NAT10 using remodelin reversed the ability of doxorubicin to induce the EMT in HCC cells. The mechanism by which NAT10 regulates the EMT and doxorubicin sensitivity in HCC cells merits further investigation

Objectives

Methods

Results

Discussion

Conclusion