Elsevier

Biomedicine & Pharmacotherapy

Volume 91, July 2017, Pages 803-811
Biomedicine & Pharmacotherapy

MiR-330 inhibits IL-22-induced keratinocyte proliferation through targeting CTNNB1

https://doi.org/10.1016/j.biopha.2017.05.005Get rights and content

Abstract

Psoriasis is a common chronic inflammatory skin disease which is characterized by hyperproliferation and aberrant differentiation of keratinocytes; however the exact pathogenesis is largely unknown. Interleukin-22 (IL-22) has demonstrated its vital role in T cell-mediated immune response by interacting with keratinocytes in the pathogenesis of psoriasis. The microRNAs (miRNAs) are a class of small non-coding RNA molecules that play important roles in cellular processes by regulating gene expression at the post-transcriptional level. MiR-330 has been reported to inhibit the proliferation and migration of mouse keratinocytes. In the present study, we indicated that miR-330 expression in lesion tissue of psoriasis patients was specifically down-regulated, and could inhibit IL-22-induced proliferation of HaCaT and HKC cell. Wnt/β-catenin pathway plays an essential role in the pathogenesis of psoriasis. By direct targeting CTNNB1, miR-330 could significantly downregulate IL-22-induced CTNNB1 expression. In addition, we found that the downstream targets of β-catenin, CyclinD1 and Axin2, could be affected by miR-330; miR-330 could suppress CyclinD1 protein expression and rescue Axin2 protein expression. Taken together, we indicated miR-330 inhibits IL-22-induced proliferation of HaCaT and HKC cell by targeting CTNNB1 and subsequently affect the downstream factors, CyclinD1 and Axin2 for the first time, and provide diagnostic markers and a novel target for psoriasis treatment.

Introduction

Psoriasis is a chronic, immune-mediated inflammatory skin disease which is characterized by abnormal hyperplasia and differentiation in keratinocytes, resulting in histopathologic changes, including thickening of the epidermis and parakeratosis [1], [2], [3]. Even though the underlying mechanisms remain largely elusive, immunological dysfunction plays a crucial role in the process of this disease. In particular, immune response mediated by T cells and keratinocytes has participated in the initiation and maintenance of psoriasis [3]. Numerous inflammatory cytokines produced by immune cells contribute to the proliferation of keratinocytes [4]. Interleukin-22 (IL-22), belonging to the IL-10 superfamily [5], is mainly produced by activated Th1, Th17, Th22 and acts only on keratinocytes rather than immune cells in the skin [6], [7], [8]. The expression of IL-22 mRNA is massively increased in lesions and serum from psoriatic patients and it is correlated with the disease severity [9], [10]. In addition, IL-22 promotes keratinocyte proliferation, induces keratinocyte migration and down-regulates the expression of genes associated with keratinocyte differentiation in vitro [11], [12]. All evidence strongly suggests that IL-22 plays a critical role in the pathogenesis of psoriasis.

MicroRNAs (miRNAs) are small single-stranded noncoding RNAs that play important roles in the physiological and pathological processes in human beings. They can negatively regulate the expression of target genes at the post-transcriptional level by binding to the 3’untranslated region (UTR) of the target mRNA [13]. Recent studies have demonstrated that altered miRNA profiles were linked to the pathogenesis of psoriasis, thus opening up new horizons for the research on psoriasis [14], [15], [16], [17], [18].

The Wnt/β-catenin pathway is best known for its role in embryogenesis and development [19], [20]. Some researchers highlight that the Wnt/β-catenin pathway is also involved in the regulation of cancer cell proliferation [21], [22]. In addition, β-catenin has been reported to be promoted keratinocyte proliferation [23], [24]. In the present study, we investigated the functional role of miR-330 in the regulation of IL-22 induced keratinocyte proliferation. Further, we investigated whether miR-330 exerts its functions through β-catenin signaling and the downstream target factors. Taken together, these findings provided a novel molecular basis for the potential effect of miR-330 in the treatment of psoriasis.

Section snippets

Clinical specimens

With the approval of the Ethic Committee of The Second Affiliated Hospital of Hunan University of Chinese Medicine, we collected 26 paired psoriatic skin lesional tissues and normal non-lesional tissues from December 2014 to June 2016. 3 mm punch biopsies were taken from lesional skin. The non-lesional skin was taken about 5 cm away from the lesion biopsy. All psoriasis patients were diagnosed by dermatologists according to diagnostic standard for psoriasis. All cases signed informed consent. All

Expression of miR-330 in psoriatic keratinocytes

The disorders of miRNAs have been frequently reported [25], [26], [27]. Kim et al. revealed that miR-330-5p inhibits proliferation and migration of keratinocytes by targeting Pdia3 expression [25]. Here, we firstly evaluated the expression levels of miR-330 in 26 paired of psoriatic lesion tissues and non-lesion tissues using real-time PCR assays. Results showed that miR-330 expression was significantly down-regulated in lesion tissues, compared to non-lesion tissues (Fig. 1A, P < 0.01). Given

Discussion

Studies have indicated that small variation in miRNA expression profiles can lead to important biological alterations [35]. Dysfunctions of miRNAs in the pathogenesis of psoriasis have been frequently reported [16], [18], [36]. Disordered miRNAs might mediate the hyperproliferation of keratinocyte in psoriasis vulgaris [16]. Previously, miR-330 has been reported to inhibit the proliferation and migration of keratinocytes by targeting Pdia3 expression [25]. Through direct targeting the 3′UTR of

Conflict of interest

All authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Acknowledgement

This study was funded by National Natural Science Foundation of China (81273769).

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