Original ArticlesADAM9 mediates the interleukin-6-induced Epithelial–Mesenchymal transition and metastasis through ROS production in hepatoma cells
Introduction
Hepatocellular carcinoma (HCC) is the third leading cause of worldwide cancer-related deaths [1]. Because of the high percentage of tumor recurrences and metastases following curative resections, it is important to understand the mechanisms of HCC metastasis and to identify new therapeutic targets. The epithelial–mesenchymal transition (EMT), in which epithelial cancer cells lose their polarity and become motile mesenchymal cells, plays a pivotal role in the dissemination of malignant hepatocytes during HCC progression [2]. Moreover, an increasing number of important EMT-related transcription factors, including Snail, Slug, Twist, and Zeb1/2 have been reported to activate the EMT during tumor progression [3].
Inflammation is a common characteristic of cancers. An increasing number of epidemiological and experimental studies have reported that inflammation plays an important role in promoting invasion and metastasis of human cancers [4,5]. In particular, interleukin 6 (IL-6) is an important regulator linking chronic inflammation to HCC progression [6]. IL-6 levels are elevated in the serum and cancer tissues of patients with HCC, and the levels correlate with tumor metastasis and patient survival [7]. Recent studies have also reported that IL-6 was capable of inducing an EMT phenotype in human liver cancer cells [8,9].
A disintegrin and metalloprotease (ADAM) is a modular type I transmembrane protein that contains a metalloprotease and a disintegrin-like domain. Recent studies have reported a relationship between increased levels of ADAM and cancer progression [10,11]. Twenty-one members of the ADAM family have been identified in humans, 13 of which have functional protease activities. ADAM9 possesses potent biological activities, is highly expressed in cancers including HCC and cancers of the prostate, pancreas, breast, and colon, and is associated with cancer progression and poor clinical outcomes [[11], [12], [13], [14]]. Many studies have also reported that ADAM9 is expressed at the invasion edges of several cancers, and has the ability to degrade specific extracellular matrix substrates, release active growth factors, and interact with key regulatory factors, which suggests that ADAM9 regulates several cancer-related processes, including cell growth, apoptosis, and invasion [[15], [16], [17]].
Increased knowledge of the mechanism involving the IL-6-induced EMT in HCC is needed. In the present report, we described the first evidence that induced expression of ADAM9 mediates this process. We also identified a crucial role of ADAM9 in HCC cell invasion, lung metastasis, and Snail-mediated E-cadherin suppression. Furthermore, we showed a positive correlation between ADAM9 and IL-6 levels and Snail expression in HCC samples. Taken together, our data indicated that the overexpression of ADAM9, stimulated by IL-6, is a strong inducer of the EMT, and may therefore serve as a therapeutic target.
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Cell lines and animals
MHCC97H cells, which have high malignant potential, were established at the Liver Cancer.
Institute of Fudan University [18]. Huh7 cells, which have low malignant potential, were obtained from the Chinese Academy of Sciences (Shanghai, China). Both hepatoma cell lines were routinely maintained.
Male BALB/c nude mice (4–6 weeks old; Shanghai SLAC Laboratory Animal Co., Shanghai, China) were raised in specific pathogen-free conditions. All animal protocols were approved by the Ethical Committee on
IL-6 promotes HCC metastasis and invasion
We screened different concentrations of soluble IL-6 in Huh7 and MHCC97H cells to identify the maximal response of genes associated with invasion and metastasis (MMP2, MMP9, SPP1, and CD44). Gene expression levels in Huh7 cells were altered following incubation with IL-6, and significant differences were found at concentrations of 10 and 25 ng/mL (Fig. 1A). The expression levels were also significantly elevated in MHCC97H cells using soluble IL-6 concentrations of 10 and 25 ng/mL (Fig. 1A).
Discussion
HCC is an inflammation-related cancer that often develops in association with chronic liver inflammation caused by hepatitis B or hepatitis C virus infections [31]. IL-6 is recognized as a major mediator involved in the regulation and maintenance of the inflammatory response [32]. Previous studies suggested that IL-6 levels increase in liver cancer and that serum IL-6 levels are associated with poor prognoses in liver cancer [[7], [8], [9]]. In the present study, our objective was to determine
Authors' contributions
Yinying Dong, Zhifeng Wu and Mingyan He performed the research, analyzed data, and participated in writing this manuscript. Yuhan Chen, Yixing Che, Xiaoyun Shen, Li Zhang and Li Zhang participated in the conduction of this study. Zhaochong Zeng designed this study, provided funding, and gave the final approval for the submission of this paper for publication.
Conflicts of interest
The authors declare no potential conflicts of interest.
Acknowledgements
This work was supported by the National Natural Science Foundation of China (Grant No. U1505229). We are very grateful to Ke Qiao for the excellent technical assistance.
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2022, GeneCitation Excerpt :In this study, 26 genes were identified to be potential regulators in HCC metastasis via WGCAN, differentially expressed gene analysis, and gene enrichment analysis (Figs. 2–4, Table S2). Among these 26 genes, 20 genes (ALDOA(Li et al., 2019b), BSG(Yang et al., 2021), CLIC1(Peng et al., 2021), ENO1(Jiang et al., 2020), HSP90AB1(Meng et al., 2018), DNAJB1(Kastenhuber et al., 2017), IGF1(Xu et al., 2019), ITGA6(Duan et al., 2021), PPP1CA(Xiang et al., 2021), RAN(Yao et al., 2019), TSPAN8(Fang et al., 2016), TMPO(Guo and Wang, 2020), VWF(Mochizuki et al., 2012), TAGLN2(Shi et al., 2020), RUVBL1(Mello et al., 2020), ADAM9(Dong et al., 2018), CCNB2(Li et al., 2019a), NDRG1(Cheng et al., 2011), SCRIB(Wan et al., 2018), ANLN(Jia et al., 2021)) were previously identified to be functionally involved in HCC metastasis. Our results verified the previous reports which indicates that the method used in this study is feasible and effective.
Systematic review of the roles of interleukins in hepatocellular carcinoma
2020, Clinica Chimica ActaCitation Excerpt :ADAM9 and NOX1 (one of the homologues of gp91phox) interact to promote ROS production, followed by enhanced Snail expression. Because ADAM9 regulates HCC metastasis and poor prognosis by increasing Snail expression and promoting EMT by processes involving ROS production, it is probable that overexpression of ADAM9 stimulated by IL-6 is a strong inducer of EMT [44]. In response to IL-6 stimulation, increased progranulin (PGRN) expression from HCC cells is via MEK/Erk signaling activating transcriptional factor C/EBPβ.
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These authors contributed equally to this work.