Primate-specific miRNA-637 inhibited tumorigenesis in human pancreatic ductal adenocarcinoma cells by suppressing Akt1 expression

doi:10.1016/j.yexcr.2018.01.026

Experimental Cell Research

Volume 363, Issue 2, 15 February 2018, Pages 310-314

https://doi.org/10.1016/j.yexcr.2018.01.026 Get rights and content

Highlights

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miR-637 acted as a tumor suppressor in pancreatic cancer.
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Akt1 was a real target of miR-637 in pancreatic ductal adenocarcinoma (PDAC) cells.
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Akt1 knockdown induced the cell growth inhibition and apoptosis in PDAC cells.

Abstract

As a primate-specific microRNA, miR-637 has been discovered for nearly 10 years. Our previous study demonstrated that miR-637 acted as a suppressor in hepatocellular carcinoma. However, its biomedical significance in pancreatic cancer remains obscure. In the present study, miR-637 was found to be significantly downregulated in pancreatic ductal adenocarcinoma (PDAC) cell lines and most of the PDAC specimens. Furthermore, the enforced overexpression of miR-637 dramatically inhibited cell proliferation and induced apoptosis of PDAC cells. Akt1, as a serine/threonine-protein kinase, has been identified as an oncogene in multiple cancers including pancreatic cancer. Our data confirmed that Akt1 was a novel target for miR-637, and its knockdown also induced cell growth inhibition and apoptosis in PDAC cells. In conclusion, our data indicated that miR-637 acted as a tumor-suppressor in PDAC, and the suppressive effect was mediated, at least partially, by suppressing Akt1 expression.

Introduction

Pancreatic cancer is the fourth leading cause of cancer deaths, and the most common type is pancreatic ductal adenocarcinoma (PDAC). As one of the most lethal malignancies, it is responsible for hundreds of thousands of deaths each year [1], [2]. Recent advances in therapeutic strategies have improved the quality of the patients with early-stage PDAC [3]. However, most patients were detected with advanced stages and the 5-year survival rate remains about 6% [4]. Thus, further investigations are urgently needed to find the potential therapeutic targets for PDAC patients.

MicroRNAs are a family of small non-coding RNAs which act as gene regulators in various biological activities [5]. They modulate the target gene expression by repressing translation or regulating mRNA degradation via binding to the 3′UTR of their target genes [6]. It is well known that miRNAs involve in human carcinogenesis and aberrantly expressed miRNAs play essential roles in pancreatic cancer progression [7]. Several miRNAs have been considered as potential biomarkers for pancreatic cancer, such as miR-21, miR-155, miR-196a, and miR-210, which were shown to be upregulated in cancer tissues [8], [9], [10], serum [11], [12], [13], [14], fecal specimens [15], [16] and pancreatic juices [17], [18] derived from pancreatic cancer patients. Other miRNAs have been reported to directly regulate cell proliferation and survival, i.e. miR-216 [19], miR-372 [20], miR-195 [21], miR-186 [22] were downregulated in pancreatic cancer specimens and served as suppressors in tumorigenesis. As for miR-637, our previous study showed that it inhibited cell proliferation and tumorigenesis of hepatocellular carcinoma [23]. However, the role of miR-637 in pancreatic cancer remains elusive.

In the present study, miR-637 was found to be downregulated in PDAC cells and clinical specimens. Further investigation revealed that miR-637 overexpression suppressed cell proliferation and induced apoptosis of PDAC cells via directly suppressing oncogene Akt1 expression. Therefore, miR-637 may be a promising molecular target for PDAC patients.

Section snippets

Cell culture

A panel of human PDAC cell lines including SW1990, BxPC-3 and Capan-2 and the pancreatic duct epithelial cells (HPDE) were cultured in Dulbecco's modified Eagle's medium (DMEM, Invitrogen) supplemented with 10% FBS and 1% Penicillin-Streptomycin.

Clinical samples

Twenty-five paired primary PDAC specimens and matched adjacent non-tumor tissues were collected from tumor resections at the Eighth Affiliated Hospital, Sun Yat-sen University and the people's Hospital of Baoan Shenzhen. All tissues were confirmed by

MiR-637 was decreased in human PDAC cells and most clinical specimens

Previous studies have demonstrated that miR-637 expression was decreased in hepatocellular carcinoma [23], and our results corroborated that it was significantly downregulated in most PDAC cells (Fig. 1A). Moreover, we also found that miR-637 was decreased in PDAC tissues compared to their adjacent non-tumor tissues and the normal PDAC tissues (Fig. 1B). Therefore, miR-637 downregulation is a frequent event in human PDAC, which may be involved in malignant tumor development and progression.

MiR-637 overexpression induced growth inhibition and apoptosis in PDAC cells

Discussion

As a tumor suppressor, miR-637 has been demonstrated to be associated with several types of human cancers, such as hepatocellular carcinoma [23], breast cancer [24], follicular thyroid carcinomas [25] and glioma [26]. However, its functions in pancreatic cancer have not been completely clarified. In the present study, we identified miR-637 as a putative tumor suppressor in PDAC cells and tissues, and provided a basis for the potential application of miR-637 in cancer therapy.

Pancreatic cancer

Acknowledgement

This work was partially supported by grants from the National Natural Science Foundation of China (81773066 to W.F. and 81772404 to J.Z.).

Conflict of interest

We declare that there are no potential conflicts of interest.

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Non-small cell lung cancer (NSCLC) is the most common type of lung cancer with a high mortality rate and poor prognosis. miR-637 has been reported to regulate tumor progression and act as a prognosis biomarker of various cancers. Its functional role in NSCLC was investigated in this study.
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miR-637 expression was significantly decreased in NSCLC tissues and cell lines relative to normal tissues and cells. The survival rate of NSCLC patients with low miR-637 expression was lower than that of patients with high miR-637 expression. Additionally, miR-637 served as a tumor suppressor that inhibited cell proliferation, migration, and invasion of NSCLC.
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It functions as a tumor suppressor via targeting and downregulating NUPR1 expression, which was shown to lead to the inhibition of proliferation, migration, and invasion in colorectal cancer cells.34 Moreover, supporting our previous results, miR-637 was found to target Akt1 and hamper tumorigenesis in both thyroid carcinoma and pancreatic ductal adenocarcinoma.35,36 miR-637 could also inhibit Akt phosphorylation and promote melanoma progression.37
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Additionally, our research demonstrated that lncRNA FAL1 promotes CRC tumorigenesis as the target ceRNA of miR-637, thus indirectly regulating the downstream targets of miR-637. Studies have demonstrated that miR-637 was downregulated and functioned as a tumour suppressor in several cancers (Que et al., 2015; Xu et al., 2018; Yuan et al., 2018; Zhang et al., 2011). In this study, we also identified that miR-637 inhibited tumorigenesis of CRC cells, and the expression level of miR-637 presented a negative correlation with lncRNA FAL1, further suggested that lncRNA FAL1 could sponge miR-637 to promote cell proliferation and invasion in CRC.
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Primate-specific miRNA-637 inhibited tumorigenesis in human pancreatic ductal adenocarcinoma cells by suppressing Akt1 expression

Highlights

Abstract

Introduction

Section snippets

Cell culture

Clinical samples

MiR-637 was decreased in human PDAC cells and most clinical specimens

MiR-637 overexpression induced growth inhibition and apoptosis in PDAC cells

Discussion

Acknowledgement

Conflict of interest

Cancer Treat. Rev.

Cell

J. Gastrointest. Surg.

Transl. Oncol.

Cell Signal.

Cancer statistics, 2015

CA Cancer J. Clin.

Pancreatic adenocarcinoma

New Engl. J. Med.

Genetics and biology of pancreatic ductal adenocarcinoma

Genes Dev.

The functions of animal microRNAs

Nature

Current and future biomarkers for pancreatic adenocarcinoma

Tumour Biol.