Tzu Chi Medical Journal

: 2020  |  Volume : 32  |  Issue : 1  |  Page : 1--4

The role of hydrogen peroxide-inducible clone-5 in tumor progression

Wen-Sheng Wu 
 Department of Laboratory Medicine and Biotechnology, College of Medicine, Tzu Chi University, Hualien, Taiwan

Correspondence Address:
Prof. Wen-Sheng Wu
Department of Laboratory Medicine and Biotechnology, College of Medicine, Tzu Chi University, 701, Section 3, Jhongyang Road, Hualien


The poor prognosis of cancers such as hepatocellular carcinoma is due to high recurrence rate mainly caused by metastasis. Target therapy aiming at critical signal molecules within these pathways is one of the promising strategies for the prevention of metastasis. Hydrogen peroxide-inducible clone-5 (Hic-5), which belongs to the paxillin superfamily, is emerging as a potential target along the metastatic signaling pathway. Hic-5 and paxillin share similar structural features; however, there are a lot of different biochemical properties between them, including tissue-specific distribution, regulation of gene expression, critical signal cascade, and the impacts on cellular phenotypes. This review focus on the recent studies of Hic-5 related to its impacts on signal transduction and transcription responsible for tumor progression. Hic-5 may regulate mitogen-activated protein kinase cascade for cell migration and invasion in various systems. Hic-5 can mediate transforming growth factor-β1-induced epithelial–mesenchymal transition (EMT) via RhoA- and Src-dependent signaling. Moreover, Hic-5 plays a central role in a positive feedback Hic-5-NADPH oxidase-ROS-JNK signal cascade. This sustained signaling is required for regulating EMT-related genes including E-cadherin, Snail, MMP9, and Zeb-1. In addition, Hic-5 can be a transcription coregulatory factor for a lot of nuclear receptors. Owing to the critical role of Hic-5 in signal transduction and transcription responsible for tumor progression, it can be a potential therapeutic target for the prevention of tumor metastasis.

How to cite this article:
Wu WS. The role of hydrogen peroxide-inducible clone-5 in tumor progression.Tzu Chi Med J 2020;32:1-4

How to cite this URL:
Wu WS. The role of hydrogen peroxide-inducible clone-5 in tumor progression. Tzu Chi Med J [serial online] 2020 [cited 2020 Sep 18 ];32:1-4
Available from:

Full Text


Tumor metastasis is responsible for the high recurrence rate and poor prognosis of cancers such as hepatocellular carcinoma (HCC) [1]. Metastasis is initiated by the epithelial–mesenchymal transition (EMT) which facilitates migration and invasion of primary tumor. This will promote the entrance of tumor cells into the blood vessel (intravasation). The tumor cells that survive in the circulation will move out of the blood vessel (extravasation) and finally colonize at the metastatic loci. The tumor microenvironment contains a lot of growth factors and cytokines such as hepatocyte growth factor (HGF) [2] and transforming growth factor-β (TGF-β) [3] which are able to trigger the progression of a lot of tumors such as HCC via various molecular pathways. Targeting the critical molecules in the signal pathways of metastasis is one of the promising strategies of antitumor progression. Recently, hydrogen peroxide-inducible clone-5 (Hic-5), which belongs to the paxillin superfamily, is emerging as a potential target along the metastatic signaling pathway. This review focuses on the recent studies of Hic-5 related to its impacts on signal transduction and transcription responsible for tumor progression.

 Physiological Function of Hydrogen Peroxide-Inducible Clone-5 as a Focal Adhesion Adaptor

In the past decades, paxillin is known to be the most critical adaptor molecular on focal adhesion (FA) responsible for mediating signal crosstalk between integrin and metastatic factors [3]. Among the members in paxillin superfamily, Hic-5 is the most homologous to paxillin [4]. Given the structure similarity between Hic-5 and paxillin, they share many biological features, including the localization at FA and similar FA-binding factors such as protein tyrosine kinase 2β (Pyk2) and FA kinase [4]. Importantly, Hic-5 and paxillin can respond to the changes in integrin–extracellular matrix (ECM) and receptor tyrosine kinase-mediated signaling, leading to EMT and cell migration [5]. However, there are also a lot of different biochemical properties between them including tissue-specific distribution, regulations of critical signal cascade and gene expression, and the impacts on cellular phenotypes [5].

 the Role of Hydrogen Peroxide-Inducible Clone-5 in Tumor Progression

Hic-5 can be induced by a lot of tumor progressive factors such as TGF-β [4], HGF, [5] and reactive oxygen species (ROS) [6]. Previously, Hic-5 was reported to mediate TGF-β1-induced EMT and cell migration of mammary epithelial cells MCF10A via the repression of E-cadherin [7]. Pignatelli et al. further demonstrated that Hic-5 was able to promote ECM degradation and invasion that facilitate TGF-β1-induced EMT of MCF10A breast cancer cells [8]. Hic-5 can also regulate EMT of ovarian cancer cells in a TGF-β-independent manner [9]. In a more detailed cellular mechanistic study in three-dimensional ECM, Hic-5 may regulate the interchange of the amoeboid and mesenchymal phenotypes essential for the plasticity of MCF10A [10]. Recently, our study revealed that Hic-5 could be induced by HGF responsible for HCC progression and may serve as a potential HCC prognosis marker [11]. In addition, Hic-5 was also highly expressed in the cancer-associated fibroblast (CAF), required for deposition and remodeling of the stromal ECM in the tumor environment to promote noncell autonomous progression of breast [12] and colorectal cancer [13]. Moreover, Hic-5 is essential for the formation and organization of rosettes in cancer-associated fibroblasts [14]. Together, Hic-5 plays a critical role in tumor progression and may serve as a prognostic indicator of tumors such as HCC.

 the Molecular Mechanisms for Hydrogen Peroxide-Inducible Clone-5 to Trigger Tumor Progression

How Hic-5 can be a critical player in tumor progression greatly relies on its impact on signal transduction and transcription as described below.

The signaling pathway mediated by hydrogen peroxide-inducible clone-5

Hic-5 can enhance TGF-β-induced signaling by inactivating the inhibitory Smads, such as Smad3 [15] and Smad7, which facilitates cell migration. Hic-5-mediated TGF-β-induced EMT was also dependent on RhoA/ROCKI [7] and Src [8]. Hic-5 may regulate mitogen-activated protein kinase (MAPK) cascade for cell migration and invasion in various systems. For example, Hic-5 induced matrix degradation, cell migration, and invasion via the regulation of Rac1-p-38 (MAPK) pathway [8]. Hic-5 can associate with JNK (MAPK) and its upstream kinase MAPKK4 to trigger the activation of matrix metalloproteinase 2 [16]. In addition, Hic-5 enhances migration via MEK-ERK cascade during endothelial cell migration to lysophosphatidic acid [17]. Recently, it was demonstrated that Hic-5 mediated Src-induced invadopodia rosette formation and organization in active Src-transfected NIH3T3 fibroblasts and cancer-associated fibroblasts [14].

The association of Hic-5 with ROS signaling, which was well known to be critical for tumor progression [18],[19],[20], has been highlighted in recent years. Not only Hic-5 gene expression was found to be induced by ROS [21], as its name suggests, but also it has a great impact on ROS generation. Previously, Hic-5 was found to participate in ROS generation during the migration of endothelial cell [22]. In this context, Hic-5 serves as an adaptor for the assembly of FA complex required for NADPH oxidase-dependent ROS production [22]. In the vascular smooth muscle cells, Hic-5 mediated TGF-β-induced activation of NADPH oxidase required for ROS generation and cell adhesion [23]. Our recent report demonstrated that Hic-5 serves as a mediator of the ROS-JNK signaling pathway for HCC progression [11]. Initially, Hic-5 appears to locate both upstream and downstream of ROS-JNK cascade in a patient-derived HCC cell line, HCC329 [11]. Further, we found that Hic-5 plays a central role in the positive feedback Hic-5-NADPH oxidase-ROS-JNK-c-jun molecular circuit in another patient-derived HCC cell line, HCC413 (unpublished result). Briefly, in the upstream, Hic-5 can associate with the regulators of NADPH oxidase including Rac1, Traf4, and Pyk2 which are essential for the activation of NADPH oxidase and ROS generation. In the downstream, we confirm that ROS-JNK pathway, indeed, is responsible for the upregulation of Hic-5 gene expression via the transcriptional factor c-jun coupled with AP4. The Hic-5 thus induced in turn reactivates NADPH oxidase and ROS generation for sustaining JNK signaling. Taken together, a positive feedback Hic-5-Rac1-Traf4-Pyk2-NADPH oxidase-ROS-JNK/c-jun cascade was established for HCC progression [Figure 1].{Figure 1}

The impact of hydrogen peroxide-inducible clone-5 on gene transcription

There are two ways for Hic-5 to impact gene expressions in various pathophysiological processes. One of them is the effect on the transcriptional level. Previous studies have demonstrated Hic-5 to be a transcription coregulatory factor of a lot of nuclear receptors [24], such as glucocorticoid receptor (GR) [25],[26], androgen receptor [27], and progesterone receptor [28]. For example, Hic-5 may influence genomic occupancy of GR as well as transcription complex assembly, thus serving as an on/off switch for glucocorticoid regulation of many genes [25]. Furthermore, Hic-5 can be a coactivator of progesterone receptor involved in the regulation of DKK1 and calcitonin expression [28].

The other way for Hic-5 to impact gene expression is based on the signal pathway triggered by Hic-5 as described above. For example, Hic-5 can mediate gene expression of collagen and α-smooth muscle actin triggered by TGF-β/Smad pathway, leading to liver fibrosis [29] or keloids [30]. Moreover, Hic-5 was responsible for transcription of oncogenes such as c-fos [31], and overexpression of Hic-5 could increase the expression of Sp1 and survivin, which inhibited the cell apoptosis, thereby reducing the cell damage induced by hypoxia [32]. In our study, we found that Hic-5-mediated NADPH oxidase-ROS-JNK/c-jun signal cascade can regulate the expressions of EMT-related genes including the suppression of E-cadherin and upregulation of transcription factors Snail and Zeb-1 and a matrix degradation enzyme MMP9. Finally, in the tumor environment, Hic-5 promoted the expression of lysyl oxidase and collagen I in CAFs, which is responsible for orchestrating or generating a tumor-promoting stroma [13].

 Hydrogen Peroxide-Inducible Clone-5 Can Be Regarded as a Promising Therapeutic Target

Given the critical role of Hic-5 in mediating the signal transduction and transcription, the suppression of Hic-5 may benefit the prevention of disease progression. Previously, Hic-5 was regarded as a novel factor in vascular remodeling and can be a potential therapeutic target for vascular disorders [33]. Moreover,in vivo Hic-5 knockdown by siRNA repressed CCl4-induced liver fibrosis in mice [29]. The feasibility that Hic-5 can be a molecular target for blocking tumor progression was also emerging. Previously, Hic-5 has been suggested to be a possible molecular target for the treatment of melanoma [34]. Recently, we also found significant suppression of intrahepatic metastasis of a liver-transplanted patient-derived HCC cell in mice injected peritoneally with chemically modified Hic-5 siRNA (unpublished result).

In addition, one advantage of using Hic-5 as a target is its limited expression in normal tissue compared with paxillin [6]. Thus, target therapy aiming at Hic-5 may cause fewer side effects than paxillin.

 Conclusion and Perspective

As an adaptor on the FA, Hic-5 can mediate a lot of signaling pathways including the positive feedback Traf4/Pyk2-NADPH oxidase-ROS-JNK signal cascade, regulating the expression of EMT-related genes. Hic-5 can also trigger MEK-ERK, Rac1-p38, or RhoA/ROCK-Src pathway for cell migration and invasion and upregulating survivin for anti-apoptosis [Figure 1]. In future, it is worthy of investigating whether Hic-5 can be an ideal therapeutic target for the prevention of tumor progression.

Financial support and sponsorship

This work is supported by the project of Tzu Chi Medical Mission Project (TCMMP108-03-02) granted by Tzu Chi Medical Foundation.

Conflicts of interest

There are no conflicts of interest.


1Tang ZY, Ye SL, Liu YK, Qin LX, Sun HC, Ye QH,et al . Adecade's studies on metastasis of hepatocellular carcinoma. J Cancer Res Clin Oncol 2004;130:187-96.
2Goyal L, Muzumdar MD, Zhu AX. Targeting the HGF/c-MET pathway in hepatocellular carcinoma. Clin Cancer Res 2013;19:2310-8.
3Deakin NO, Turner CE. Paxillin comes of age. J Cell Sci 2008;121:2435-44.
4Brown MC, Turner CE. Paxillin: Adapting to change. Physiol Rev 2004;84:1315-39.
5Deakin NO, Turner CE. Distinct roles for paxillin and Hic-5 in regulating breast cancer cell morphology, invasion, and metastasis. Mol Biol Cell 2011;22:327-41.
6Deakin NO, Pignatelli J, Turner CE. Diverse roles for the paxillin family of proteins in cancer. Genes Cancer 2012;3:362-70.
7Tumbarello DA, Turner CE. Hic-5 contributes to epithelial-mesenchymal transformation through a RhoA/ROCK-dependent pathway. J Cell Physiol 2007;211:736-47.
8Pignatelli J, Tumbarello DA, Schmidt RP, Turner CE. Hic-5 promotes invadopodia formation and invasion during TGF-β-induced epithelial-mesenchymal transition. J Cell Biol 2012;197:421-37.
9Sheta R, Wang ZQ, Bachvarova M, Plante M, Gregoire J, Renaud MC,et al . Hic-5 regulates epithelial to mesenchymal transition in ovarian cancer cells in a TGFβ1-independent manner. Oncotarget 2017;8:82506-30.
10Gulvady AC, Dubois F, Deakin NO, Goreczny GJ, Turner CE. Hic-5 expression is a major indicator of cancer cell morphology, migration, and plasticity in three-dimensional matrices. Mol Biol Cell 2018;29:1704-17.
11Wu JR, Hu CT, You RI, Pan SM, Cheng CC, Lee MC,et al . Hydrogen peroxide inducible clone-5 mediates reactive oxygen species signaling for hepatocellular carcinoma progression. Oncotarget 2015;6:32526-44.
12Goreczny GJ, Ouderkirk-Pecone JL, Olson EC, Krendel M, Turner CE. Hic-5 remodeling of the stromal matrix promotes breast tumor progression. Oncogene 2017;36:2693-703.
13Omoto T, Kim-Kaneyama JR, Lei XF, Orimo A, Ohnishi K, Yoshihara K,et al . The impact of stromal hic-5 on the tumorigenesis of colorectal cancer through lysyl oxidase induction and stromal remodeling. Oncogene 2018;37:1205-19.
14Gulvady AC, Forsythe IJ, Turner CE. Hic-5 regulates Src-induced invadopodia rosette formation and organization. Mol Biol Cell 2019;30:1298-313.
15Wang H, Song K, Sponseller TL, Danielpour D. Novel function of androgen receptor-associated protein 55/Hic-5 as a negative regulator of Smad3 signaling. J Biol Chem 2005;280:5154-62.
16Lei XF, Kim-Kaneyama JR, Arita-Okubo S, Offermanns S, Itabe H, Miyazaki T,et al . Identification of Hic-5 as a novel scaffold for the MKK4/p54 JNK pathway in the development of abdominal aortic aneurysms. J Am Heart Assoc 2014;3:e000747.
17Avraamides C, Bromberg ME, Gaughan JP, Thomas SM, Tsygankov AY, Panetti TS. Hic-5 promotes endothelial cell migration to lysophosphatidic acid. Am J Physiol Heart Circ Physiol 2007;293:H193-203.
18Wu WS. The signaling mechanism of ROS in tumor progression. Cancer Metastasis Rev 2006;25:695-705.
19Hu CT, Wu JR, Cheng CC, Wang S, Wang HT, Lee MC,et al . Reactive oxygen species-mediated PKC and integrin signaling promotes tumor progression of human hepatoma HepG2. Clin Exp Metastasis 2011;28:851-63.
20Wu WS, Wu JR, Hu CT. Signal cross talks for sustained MAPK activation and cell migration: The potential role of reactive oxygen species. Cancer Metastasis Rev 2008;27:303-14.
21Nose K. Regulation of gene expression by active oxygen species. Yakugaku Zasshi 2002;122:773-80.
22Wu RF, Xu YC, Ma Z, Nwariaku FE, Sarosi GA Jr., Terada LS. Subcellular targeting of oxidants during endothelial cell migration. J Cell Biol 2005;171:893-904.
23Fernandez I, Martin-Garrido A, Zhou DW, Clempus RE, Seidel-Rogol B, Valdivia A,et al . Hic-5 mediates TGFβ-induced adhesion in vascular smooth muscle cells by a NOX4-dependent mechanism. Arterioscler Thromb Vasc Biol 2015;35:1198-206.
24Heitzer MD, DeFranco DB. Hic-5, an adaptor-like nuclear receptor coactivator. Nucl Recept Signal 2006;4:e019.
25Chodankar R, Wu DY, Schiller BJ, Yamamoto KR, Stallcup MR. Hic-5 is a transcription coregulator that acts before and/or after glucocorticoid receptor genome occupancy in a gene-selective manner. Proc Natl Acad Sci U S A 2014;111:4007-12.
26Yang L, Guerrero J, Hong H, DeFranco DB, Stallcup MR. Interaction of the tau2 transcriptional activation domain of glucocorticoid receptor with a novel steroid receptor coactivator, Hic-5, which localizes to both focal adhesions and the nuclear matrix. Mol Biol Cell 2000;11:2007-18.
27Heitzer MD, DeFranco DB. Hic-5/ARA55, a LIM domain-containing nuclear receptor coactivator expressed in prostate stromal cells. Cancer Res 2006;66:7326-33.
28Aghajanova L, Velarde MC, Giudice LC. The progesterone receptor coactivator Hic-5 is involved in the pathophysiology of endometriosis. Endocrinology 2009;150:3863-70.
29Lei XF, Fu W, Kim-Kaneyama JR, Omoto T, Miyazaki T, Li B,et al . Hic-5 deficiency attenuates the activation of hepatic stellate cells and liver fibrosis through upregulation of Smad7 in mice. J Hepatol 2016;64:110-7.
30Inui S, Shono F, Noguchi F, Nakajima T, Hosokawa K, Itami S. In vitro and in vivo evidence of pathogenic roles of Hic-5/ARA55 in keloids through Smad pathway and profibrotic transcription. J Dermatol Sci 2010;58:152-4.
31Kim-Kaneyama Jr., Shibanuma M, Nose K. Transcriptional activation of the c-fos gene by a LIM protein, Hic-5. Biochem Biophys Res Commun 2002;299:360-5.
32Jiang N, Xia J, Jiang B, Xu Y, Li Y. TUG1 alleviates hypoxia injury by targeting miR-124 in H9c2 cells. Biomed Pharmacother 2018;103:1669-77.
33Kim-Kaneyama JR, Lei XF, Arita S, Miyauchi A, Miyazaki T, Miyazaki A. Hydrogen peroxide-inducible clone 5 (Hic-5) as a potential therapeutic target for vascular and other disorders. J Atheroscler Thromb 2012;19:601-7.
34Noguchi F, Inui S, Nakajima T, Itami S. Hic-5 affects proliferation, migration and invasion of B16 murine melanoma cells. Pigment Cell Melanoma Res 2012;25:773-82.