OncoTargets and Therapy (Aug 2020)
Huaier Restrains Cholangiocarcinoma Progression in vitro and in vivo Through Modulating lncRNA TP73-AS1 and Inducing Oxidative Stress
Abstract
Daolin Ji,1,2 Wangyang Zheng,1,2 Peng Huang,1,2 Yue Yao,3 Xiangyu Zhong,1 Pengcheng Kang,1 Zhidong Wang,1 Guojing Shi,4 Yi Xu,1,2 Yunfu Cui1 1Department of Hepatopancreatobiliary Surgery, The Second Affiliated Hospital, Harbin Medical University, Harbin, Heilongjiang Province, People’s Republic of China; 2The Key Laboratory of Myocardial Ischemia, Harbin Medical University, Ministry of Education, Harbin, Heilongjiang Province, People’s Republic of China; 3Department of Endocrinology and Metabolism, The Second Affiliated Hospital, Harbin Medical University, Harbin, Heilongjiang Province, People’s Republic of China; 4Department of Biochemistry and Molecular Biology, Harbin Medical University, Harbin, Heilongjiang Province, People’s Republic of ChinaCorrespondence: Yunfu Cui; Yi Xu Department of Hepatopancreatobiliary Surgery, The Second Affiliated HospitalHarbin Medical University, 246, Xuefu Road, Harbin, Heilongjiang Province 150086, People’s Republic of ChinaTel +86 451-86605113Email [email protected]; [email protected]: Huaier, the fruiting body of Trametes robiniophila Murr, is a kind of traditional Chinese medicine. Recently, many studies have confirmed that Huaier has antitumor effects on various malignancies. Moreover, studies have demonstrated that long noncoding RNAs play an important regulatory role in the occurrence and progression of malignancies. Our present study was to explore whether Huaier has a potential antitumor effect in cholangiocarcinoma and reveal the relationship between lncRNAs and Huaier-induced tumor inhibition.Methods: Microarray assay was performed to identify the candidate lncRNAs regulated by Huaier. Quantitative real-time PCR was applied to assess the effect of Huaier on TP73-AS1 expression. The effect of Huaier on the cell viability, proliferation, migration and invasion was evaluated by CCK-8, colony formation, wound healing and Transwell assays, respectively. The ratio of cell apoptosis was determined using AO/EB, Hoechst 33342 and flow cytometry. The effect of Huaier on oxidative stress was revealed using DCFH-DA, mito-SOX, JC-1 probes and Western blotting. In addition, the effect of Huaier on tumor growth and metastasis was explored using subcutaneous tumor model and lung metastatic tumor model in nude mice.Results: In vitro, Huaier inhibited the proliferation, migration and invasion of cholangiocarcinoma cells by down-regulating TP73-AS1 and induced apoptosis through mitochondrial apoptotic pathway. In vivo, Huaier suppressed the growth and metastasis of cholangiocarcinoma by modulating the expression of proliferation and EMT-associated proteins.Conclusion: Huaier could inhibit cell proliferation, invasion and metastasis by modulating the expression of TP73-AS1, meanwhile promote apoptosis of CCA cells through disturbing mitochondrial function, inducing oxidative stress and activating caspases in vitro. In addition, Huaier could suppress tumor growth and metastasis by regulating the expression of proliferation and EMT-related proteins. In the meantime, Huaier prolonged the survival of nude mice in lung metastatic model with acceptable drug safety.Keywords: cholangiocarcinoma, Huaier, TP73-AS1, oxidative stress, apoptosis
