N-Acetyltransferase 10 Enhances Doxorubicin Resistance in Human Hepatocellular Carcinoma Cell Lines by Promoting the Epithelial-to-Mesenchymal Transition
N-Acetyltransferase 10 Enhances Doxorubicin Resistance in Human Hepatocellular Carcinoma Cell Lines by Promoting the Epithelial-to-Mesenchymal Transition
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N-乙酰转移酶 10 通过促进上皮细胞向间质细胞的转变增强人肝细胞癌细胞系中的阿霉素耐药性
DOI：
10.1155/2019/7561879
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发表时间：
2019-07-01
影响因子：
--
通讯作者：
Liang, Xiao
中科院分区：
文献类型：
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作者：
Zhang, Xiuming;Chen, Jiang;Liang, Xiao
背景N-乙酰转移酶10（NAT 10）在肝细胞癌（HCC）中高表达，但其在化疗耐药中的作用尚不清楚。本研究的目的是探讨NAT 10是否调控肝癌的上皮-间质转化（EMT）和化疗耐药。方法用NAT 10抑制剂remodelin处理HCC细胞株Huh-7、Bel-7402、SNU 387和SNU 449，或用靶向NAT 10或Twist的小抑制RNA（siRNA）转染HCC细胞株Huh-7、Bel-7402、SNU 387和SNU 449。EMT由缺氧诱导。CCK-8测定用于定量细胞活力，EdU掺入测定用于评估细胞增殖。通过蛋白质印迹法评估siRNA敲低效率和上皮/间充质标志物表达。结果使用siRNA敲低NAT 10或使用remodelin抑制NAT 10增加HCC细胞系对阿霉素的敏感性;在用Twist siRNA转染的细胞中观察到类似的效果。使用重塑素抑制NAT 10也逆转了阿霉素诱导HCC细胞中EMT的能力。抑制NAT 10可逆转缺氧诱导的EMT。最后，我们证实，在小鼠肝癌异种移植模型中，多柔比星与重塑蛋白联合应用可延缓肿瘤生长并减少肿瘤细胞增殖。结论NAT 10可能通过调控EMT而参与肝癌的化疗耐药。NAT 10调节肝癌细胞EMT和阿霉素敏感性的机制值得进一步研究。
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.