Downregulation of RIG-I mediated by ITGB3/c-SRC/STAT3 signaling confers resistance to interferon-α-induced apoptosis in tumor-repopulating cells of melanoma

Background Interferon-α (IFN-α) plays a pivotal role in host antitumor immunity, and the evasion of IFN-α signaling pathway can lead to IFN-α resistance during the treatment of cancer. Although the interplay between IFN-α and tumor cells has been extensively investigated in differentiated tumor cell...

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Main Authors: Jing Liu, Fengjun Cao, Tao Xu, Yingqiu Song, Pindong Li, Mingxing Li, Haizhou Wang, Xiongjie Yu, Yuandong Yu, YunYan Tai, Ping Chen, Xiaojun Cai, Xianhe Wang, Longchao Xiang, Rui Deng, Xiufang Zhang, Liping Gao, Xuanbin Wang
Format: Article
Language:English
Published: BMJ Publishing Group 2020-05-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/8/1/e000111.full
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author Jing Liu
Fengjun Cao
Tao Xu
Yingqiu Song
Pindong Li
Mingxing Li
Haizhou Wang
Xiongjie Yu
Yuandong Yu
YunYan Tai
Ping Chen
Xiaojun Cai
Xianhe Wang
Longchao Xiang
Rui Deng
Xiufang Zhang
Liping Gao
Xuanbin Wang
author_facet Jing Liu
Fengjun Cao
Tao Xu
Yingqiu Song
Pindong Li
Mingxing Li
Haizhou Wang
Xiongjie Yu
Yuandong Yu
YunYan Tai
Ping Chen
Xiaojun Cai
Xianhe Wang
Longchao Xiang
Rui Deng
Xiufang Zhang
Liping Gao
Xuanbin Wang
author_sort Jing Liu
collection DOAJ
description Background Interferon-α (IFN-α) plays a pivotal role in host antitumor immunity, and the evasion of IFN-α signaling pathway can lead to IFN-α resistance during the treatment of cancer. Although the interplay between IFN-α and tumor cells has been extensively investigated in differentiated tumor cells, much less attention has been directed to tumor-repopulating cells (TRCs).Methods Three-dimentional soft fibrin matrix was used to select and grow highly malignant and tumorigenic melanoma TRCs. The regulation of integrin β3 (ITGB3)-c-SRC-STAT signaling pathway in melanoma TRCs was investigated both in vitro and in vivo. The relevant mRNA and protein expression levels were analyzed by qRT-PCR and western blot analysis. Immunoprecipitation and chromatin immunoprecipitation (ChIP) followed by qPCR (ChIP-qPCR) assays were performed to detect protein-protein and protein-DNA interactions. The clinical impacts of retinoic acid inducible gene-I (RIG-I) were assessed in melanoma datasets obtained from The Cancer Genome Atlas and Gene Expression Omnibus profiles.Results IFN-α-induced apoptosis was decreased in melanoma TRCs. Compared with conventional flask-cultured cells, IFN-α-mediated STAT1 activation was diminished in melanoma TRCs. Decreased expression of RIG-I in melanoma TRCs led to diminished activation of STAT1 via enhancing the interaction between Src homology region 2 domain-containing phosphatase-1 and STAT1. In addition, low expression levels of RIG-I correlated with poor prognosis in patients with melanoma. STAT3 was highly phosphorylated in TRCs and knockdown of STAT3 reversed the downregulation of RIG-I in TRCs. Knockdown of STAT3 resulted in STAT1 activation and increased expression of the pro-apoptosis genes in IFN-α-treated TRCs. Combined treatment of STAT3 inhibitor and IFN-α increased the apoptosis rate of TRCs. Disruption of ITGB3/c-SRC/STAT3 signaling pathway significantly elevated the efficiency of IFN-α-induced apoptosis of TRCs.Conclusions In melanoma TRCs, ITGB3-c-SRC-STAT3 pathway caused RIG-I repression and then affect STAT1 activation to cause resistance to IFN-α-induced apoptosis. RIG-I is a prognostic marker in patients with melanoma. Combination of STAT3 inhibitor and IFN-α could enhance the efficacy of melanoma treatment. Our findings may provide a new concept of combinatorial treatment for future immunotherapy.
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series Journal for ImmunoTherapy of Cancer
spelling doaj-art-999d5c11cbc24000816b0c24f64a172f2024-11-08T16:30:08ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262020-05-018110.1136/jitc-2019-000111Downregulation of RIG-I mediated by ITGB3/c-SRC/STAT3 signaling confers resistance to interferon-α-induced apoptosis in tumor-repopulating cells of melanomaJing Liu0Fengjun Cao1Tao Xu2Yingqiu Song3Pindong Li4Mingxing Li5Haizhou Wang6Xiongjie Yu7Yuandong Yu8YunYan Tai9Ping Chen10Xiaojun Cai11Xianhe Wang12Longchao Xiang13Rui Deng14Xiufang Zhang15Liping Gao16Xuanbin Wang17School of Nursing and Midwifery, The University of Newcastle, Newcastle, New South Wales, Australia1 Department of Oncology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, ChinaSchool of Acu-Mox and Tuina, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan, ChinaDepartment of Radiotherapy, Cancer Hospital of China Medical University, Shenyang, Liaoning, China2 Cancer Center of Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaDepartment of Pharmacy, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, China4 Department of Gastroenterology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China1 Department of Oncology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, China1 Department of Oncology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, China1 Department of Oncology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, China1 Department of Oncology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, ChinaDepartment of Ultrasound in Medicine, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai Institute of Ultrasound in Medicine, Shanghai Jiao Tong University School of Medicine, Shanghai, China1 Department of Oncology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, China1 Department of Oncology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, China1 Department of Oncology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, China1 Department of Oncology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, ChinaDepartment of Gastroenterology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China1 Department of Oncology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, ChinaBackground Interferon-α (IFN-α) plays a pivotal role in host antitumor immunity, and the evasion of IFN-α signaling pathway can lead to IFN-α resistance during the treatment of cancer. Although the interplay between IFN-α and tumor cells has been extensively investigated in differentiated tumor cells, much less attention has been directed to tumor-repopulating cells (TRCs).Methods Three-dimentional soft fibrin matrix was used to select and grow highly malignant and tumorigenic melanoma TRCs. The regulation of integrin β3 (ITGB3)-c-SRC-STAT signaling pathway in melanoma TRCs was investigated both in vitro and in vivo. The relevant mRNA and protein expression levels were analyzed by qRT-PCR and western blot analysis. Immunoprecipitation and chromatin immunoprecipitation (ChIP) followed by qPCR (ChIP-qPCR) assays were performed to detect protein-protein and protein-DNA interactions. The clinical impacts of retinoic acid inducible gene-I (RIG-I) were assessed in melanoma datasets obtained from The Cancer Genome Atlas and Gene Expression Omnibus profiles.Results IFN-α-induced apoptosis was decreased in melanoma TRCs. Compared with conventional flask-cultured cells, IFN-α-mediated STAT1 activation was diminished in melanoma TRCs. Decreased expression of RIG-I in melanoma TRCs led to diminished activation of STAT1 via enhancing the interaction between Src homology region 2 domain-containing phosphatase-1 and STAT1. In addition, low expression levels of RIG-I correlated with poor prognosis in patients with melanoma. STAT3 was highly phosphorylated in TRCs and knockdown of STAT3 reversed the downregulation of RIG-I in TRCs. Knockdown of STAT3 resulted in STAT1 activation and increased expression of the pro-apoptosis genes in IFN-α-treated TRCs. Combined treatment of STAT3 inhibitor and IFN-α increased the apoptosis rate of TRCs. Disruption of ITGB3/c-SRC/STAT3 signaling pathway significantly elevated the efficiency of IFN-α-induced apoptosis of TRCs.Conclusions In melanoma TRCs, ITGB3-c-SRC-STAT3 pathway caused RIG-I repression and then affect STAT1 activation to cause resistance to IFN-α-induced apoptosis. RIG-I is a prognostic marker in patients with melanoma. Combination of STAT3 inhibitor and IFN-α could enhance the efficacy of melanoma treatment. Our findings may provide a new concept of combinatorial treatment for future immunotherapy.https://jitc.bmj.com/content/8/1/e000111.full
spellingShingle Jing Liu
Fengjun Cao
Tao Xu
Yingqiu Song
Pindong Li
Mingxing Li
Haizhou Wang
Xiongjie Yu
Yuandong Yu
YunYan Tai
Ping Chen
Xiaojun Cai
Xianhe Wang
Longchao Xiang
Rui Deng
Xiufang Zhang
Liping Gao
Xuanbin Wang
Downregulation of RIG-I mediated by ITGB3/c-SRC/STAT3 signaling confers resistance to interferon-α-induced apoptosis in tumor-repopulating cells of melanoma
Journal for ImmunoTherapy of Cancer
title Downregulation of RIG-I mediated by ITGB3/c-SRC/STAT3 signaling confers resistance to interferon-α-induced apoptosis in tumor-repopulating cells of melanoma
title_full Downregulation of RIG-I mediated by ITGB3/c-SRC/STAT3 signaling confers resistance to interferon-α-induced apoptosis in tumor-repopulating cells of melanoma
title_fullStr Downregulation of RIG-I mediated by ITGB3/c-SRC/STAT3 signaling confers resistance to interferon-α-induced apoptosis in tumor-repopulating cells of melanoma
title_full_unstemmed Downregulation of RIG-I mediated by ITGB3/c-SRC/STAT3 signaling confers resistance to interferon-α-induced apoptosis in tumor-repopulating cells of melanoma
title_short Downregulation of RIG-I mediated by ITGB3/c-SRC/STAT3 signaling confers resistance to interferon-α-induced apoptosis in tumor-repopulating cells of melanoma
title_sort downregulation of rig i mediated by itgb3 c src stat3 signaling confers resistance to interferon α induced apoptosis in tumor repopulating cells of melanoma
url https://jitc.bmj.com/content/8/1/e000111.full
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