The status of microRNA-21 expression and its clinical significance in human cutaneous malignant melanoma

doi:10.1016/j.acthis.2011.11.001

Acta Histochemica

Volume 114, Issue 6, October 2012, Pages 582-588

https://doi.org/10.1016/j.acthis.2011.11.001 Get rights and content

Abstract

Dysregulation of microRNA-21 plays critical roles in tumor initiation and progression. The purpose of this study was to investigate the status of microRNA-21 expression in human cutaneous malignant melanoma and determine its clinical significance. TaqMan^® real-time RT-PCR assay was performed to examine the expression of microRNA-21 in 10 cases of dysplastic nevi, 86 cases of primary cutaneous melanomas, 10 cases of melanoma metastases. The correlation of microRNA-21 expression with clinicopathological factors or prognosis of patients with cutaneous melanoma was statistically analyzed. Additionally, the effects of microRNA-21 expression on growth, apoptosis and chemo- or radiosensitivity of melanoma cells were also investigated by transfection of microRNA-21 inhibitor. We firstly showed that increased levels of microRNA-21 expression were shown from dysplastic nevi to primary cutaneous melanomas to melanoma metastases. Moreover, high miR-21 expression was found to be correlated with Breslow thickness and advanced clinical stage. Patients with high microRNA-21 expression showed shorter 5-year disease-free or overall survival than those with low microRNA-21 expression. Furthermore, multivariate regression analysis showed that the status of microRNA-21 expression was an independent prognostic factor for overall survival of patients. Antisense-mediated microRNA-21 inhibition could significantly suppress growth, increase apoptosis and enhance chemo- or radiosensitivity of human cutaneous melanoma cells by inducing the increased Bax/Bcl-2 ratio. Thus, the status of microRNA-21 might be an independent prognostic factor for patients with cutaneous melanoma, and microRNA-21 has the potential of being a novel molecular target for the treatment of human cutaneous melanoma.

Introduction

Melanoma is a malignancy of pigment-producing cells melanocytes located predominantly in the skin, but also found in the eyes, ears, leptomeninges, and oral and genital mucous membranes, and accounts for 80% of deaths arising from skin cancer (Miller et al., 2006). Malignant melanoma is highly characteristic of aggressive invasion, early metastasis and resistance to chemotherapy or radiotherapy, which results in the increased incidence and mortality worldwide, especially among white populations during past decades (Terando et al., 2003). Therefore, better understanding of the molecular mechanisms about malignant melanoma tumorigenesis and progression will be helpful to explore novel therapeutic agents and prognostic markers in the treatment of patients with cutaneous malignant melanoma.

MicroRNAs (miRNAs) are short single-stranded non-coding RNA sequences, 20–22nt in length, that play an important role in post-transcriptional gene regulation by interaction with complementary sites in the 3′-UTR of target mRNAs (Lai, 2002). Several hundred miRNAs have been cloned or predicted, but a functional role for these in many cellular processes is not fully elucidated. MiRNAs have been reported to play a vital role in many human physiological and pathological processes including development, differentiation, cellular proliferation, and apoptosis (Jovanovic and Hengartner, 2006, Tsuchiya et al., 2006, Belver et al., 2011). Hence, dysregulation of miRNAs might lead to a variety of human disorders (Braun et al., 2011, Muntoni et al., 2011, Sayed and Abdellatif, 2011). Recently, the associations of miRNAs with human cancers are also well documented (McManus, 2003). MiRNAs may function as tumor suppressors or oncogenes by targeting oncogenes or tumor suppressor genes (Shenouda and Alahari, 2009). As an oncomir, miR-21 is a well-known modulator of cell proliferation and viability in many different cell types. The overexpression of miR-21 has been found in many human cancers such as pancreatic cancer, breast cancer, colorectal cancer, lung cancer, gastric cancer (Dillhoff et al., 2008, Markou et al., 2008, Huang et al., 2009, Motoyama et al., 2010, Shibuya et al., 2010). Also, it has been reported that miR-21 could modulate proliferation, invasion, and chemosensitivity of human tumor cells (Pan et al., 2011). However, the status of miR-21 expression in cutaneous malignant melanoma and its clinical significance is still unknown.

In this study, we show for the first time that expression levels of miR-21 are frequently upregulated in cutaneous malignant melanoma. Also, high levels of miR-21 are correlated with advanced tumor stage, degree of invasion and tumor recurrence of patients. Furthermore, cutaneous malignant melanoma patients with high miR-21 expression were found to have significantly worse prognosis after potentially curative resection, and the status of miR-21 expression might be an independent prognostic factor by multivariate COX regression analysis. Additionally, we show that antisense-mediated knockdown of miR-21 can inhibit growth, induce apoptosis enhancement and increase chemo- or radiosensitivity of malignant melanoma cells.

Section snippets

Patient and tissue samples

A total of 10 cases of dysplastic nevi, 10 cases of melanoma metastases and 86 cases of primary cutaneous melanoma tissue samples were collected directly from surgery after removal of the necessary amount of tissue for routine pathology examination in the Department of Pathology, Fourth Military Medical University between 1998 and 2003. The histological diagnosis, Breslow thickness and Clark level were re-examined from 1 to 5 original sections of the primary tumor by the same pathologist who

Results

Firstly, TaqMan^® real-time RT-PCR assay was performed to examine the expression of miR-21 in 10 cases of dysplastic nevi, 10 cases of primary cutaneous melanomas, 10 cases of melanoma metastases. As shown in Fig. 1, there were significant differences in the pattern of relative miR-21 expression, with increased levels of expression from dysplastic nevi to primary cutaneous melanomas to metastatic cutaneous melanomas. Significant differences for miR-21 expression were shown between dysplastic

Discussion

There is increasing evidence that dysregulation of miRNAs plays critical roles in cancer initiation and progression, suggesting that miRNAs may play roles as tumor suppressor genes or oncogenes (Koturbash et al., 2011). Consequently, researchers are attempting to exploit and identify miRNAs that may serve as either diagnostic or prognostic markers or therapeutic targets in many different tumor types (Shah et al., 2010, Nana-Sinkam and Croce, 2011). Although the functions of some miRNAs in a

Acknowledgement

We are grateful to everyone of the Department of Pathology, the Fourth Military Medical University for their sincere help.

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¹: These authors contributed equally to this work.

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The status of microRNA-21 expression and its clinical significance in human cutaneous malignant melanoma

Abstract

Introduction

Section snippets

Patient and tissue samples

Results

Discussion

Acknowledgement

Gemcitabine sensitivity can be induced in pancreatic cancer cells through modulation of miR-200 and miR-21 expression by curcumin or its analogue CDF

Cancer Res

MicroRNA control of lymphocyte differentiation and function

Curr Opin Immunol

Knockdown of microRNA-21 inhibits proliferation and increases cell death by targeting programmed cell death 4 (PDCD4) in pancreatic ductal adenocarcinoma

Gastrointest Surg

Hyaluronan-CD44 interaction with protein kinase (Cepsilon) promotes oncogenic signaling by the stem cell marker Nanog and the production of microRNA-21, leading to down-regulation of the tumor suppressor protein PDCD4, anti-apoptosis, and chemotherapy resistance in breast tumor cells

J Biol Chem

Transcriptional mechanisms regulating skeletal muscle differentiation, growth and homeostasis

Nat Rev Mol Cell Biol

The PTEN/PI3K/AKT signalling pathway in cancer, therapeutic implications

Curr Cancer Drug Targets

MicroRNA-34b and MicroRNA-34c are targets of p53 and cooperate in control of cell proliferation and adhesion-independent growth