lncRNA OSER1-AS1 acts as a ceRNA to promote tumorigenesis in hepatocellular carcinoma by regulating miR-372-3p/Rab23 axis

doi:10.1016/j.bbrc.2019.10.105

Biochemical and Biophysical Research Communications

Volume 521, Issue 1, 1 January 2020, Pages 196-203

https://doi.org/10.1016/j.bbrc.2019.10.105 Get rights and content

Highlights

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The role of OSER1-AS1 in HCC was first explained.
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OSER1-AS1 may play an oncogenic role in HCC.
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OSER1-AS1 functions as a sponge for miR-372-3p to promote Rab23 expression.
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Rab23 is a novel target for miR-372-3p.

Abstract

Long non-coding RNAs (lncRNAs) are crucial regulators of tumorigenesis and progression in human cancer, including hepatocellular carcinoma (HCC). However, the role of most lncRNAs that are dysregulated in HCC remains to be elucidated. Here, we investigated the role of OSER1-AS1 in the progression of HCC. The results of database and qRT-PCR analysis demonstrated that OSER1-AS1 was highly expressed in HCC tissues and the high expression of OSER1-AS1 was closely associated with larger tumor size, advanced tumor stages, lower disease free survival and overall survival of HCC patients. OSER1-AS1 knockdown significantly inhibited the proliferation, invasion and migration of HCC cells, and induced the apoptosis. In addition, the dual luciferase reporter assay directly demonstrated that OSER1-AS1 functioned as a molecular sponge for miR-372-3p to promote Rab23 expression. Moreover, the results of immunohistochemistry and western blot analysis showed that Rab23 was highly expressed in HCC tissues, and the high expression of Rab23 was closely associated with the poor overall survival of HCC patients. Immunofluorescence assay also found the subcellular localization of Rab23 in HCC cells. Rab23 was obviously downregulated in cells that were transfected with miR-372-3p mimics. MiR-372-3p mimics significantly inhibited the proliferation and invasion of HCC cells). Rab23 restoration partially reversed miR-372-3p-induced tumor suppressive effects on HCC cells. In conclusion, we found that OSER1-AS1 acted as a ceRNA to sponge miR-372-3p, thereby positively regulating the Rab23 expression and ultimately acting as a tumor suppressor gene in HCC progression.

Introduction

Hepatocellular carcinoma (HCC) is the prevalent malignancy, recognized as the fourth leading cause of incidence and the second leading cause of mortality among all types of human cancers in China [1]. Although continued improvement in HCC diagnosis and therapy has been made, the 5-year survival rate of HCC patients remains dismal [2]. The high frequency of recurrence and metastasis following surgery lead to poor prognosis of HCC [3]. Therefore, it is vital to further understand the molecular mechanism of HCC progression.

Long non-coding RNAs (lncRNAs) are transcripts containing 200 nucleotides and possess no or limited protein coding function [4]. Increasing studies demonstrate that lncRNAs are crucial regulators of tumorigenesis and progression in human cancer [4]. LncRNAs could serve as a competitive endogenous RNA (ceRNA) to sponge microRNA (miRNA), indirectly regulating target genes. MiRNAs could silence gene expression through targeting 3′ untranslated regions (3′UTR) of mRNAs [5]. For example, in HCC, lncRNA SOX9-AS1 drove progression and metastasis by regulating miR-5590-3p/SOX9 axis [6]. lncRNA EIF3J-AS1 regulated CTNND2 by targeting miR-122-5p to promote HCC [7].

In present study, we confirmed the high expression of lncRNA OSER1-AS1 in HCC tissues, and examined the effect of OSER1-AS1 knockdown on HCC cell proliferation, apoptosis, migration and invasion. Furthermore, we found that OSER1-AS1 contributed to HCC progression by targeting miR-372-3p/Rab23 axis.

Section snippets

Clinical samples

Paired HCC and corresponding adjacent normal tissues were acquired from 34 cases of HCC patients who underwent pneumonectomy in Qilu Hospital. The pathological type of each tumor sample was confirmed by experienced pathologists. Fresh samples were immediately frozen in the liquid nitrogen. None of the patients had received any adjuvant chemotherapy or radiotherapy before surgery. The study was approved by the Ethical Committee of Qilu Hospital, and each patient signed the informed written

OSER1-AS1 is highly expressed in HCC

We firstly quantified the expression of OSER1-AS1 between HCC and adjacent non-tumor tissues via database analysis and qRT-PCR. The data from GEPIA [8] revealed the higher levels of OSER1-AS1 in HCC tissues compared to normal liver tissues (Fig. 1A). In additional, it demonstrated that the high expression of OSER1-AS1 was closely associated with lower disease free survival and overall survival of HCC patients (P < 0.05, Fig. 1B and C). We also clinically collected 34 paired HCC and adjacent

Discussion

HCC which accounts for about 90% of primary liver cancers witnesses low 5-year survival rate. Identifying new biomarkers for HCC diagnosis, prognosis and therapy are necessary to improve its status. LncRNAs play a key role in this process. For example, H19 is the first non-coding RNA found in HCC [9], and contributes to carcinogenesis [10]. HOTAIR also is overexpressed in HCC and is closely related to tumor progression and recurrence [11,12]. In fact, only a small fraction of lncRNAs in our

Acknowledgements

This study was supported by grants from the National Science and Technology Major Special Project of China (2017ZX09301058002), the Youth Foundation for Scientific and Technological Research in Colleges and Universities of Hebei Province, China (Grant No. QN2017054), the Shandong Provincial Natural Science Foundation, China (No. ZR2018MC008) and the National Natural Science Foundation of China (No 81971088).

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Accumulating evidence has unfolded aberrantly expressed long non-coding RNAs (lncRNAs) in the blood of cancer sufferers and exhibit significant potential as non-invasive tumor markers [4]. LncRNAs refer to a novel class of noncoding RNA longer than 200 nucleotides [5], and have been suggested to be of vital functionality in tumor development and progression [6,7]. For instance, knockdown of MALAT1 exerts inhibitory action in the progression of anaplastic thyroid carcinoma [8].
Abnormal long non-coding RNAs (lncRNAs) have been detected in esophageal squamous cell carcinoma (ESCC). Here, we focused on lncRNA ZNF667-AS1 and its downstream mechanism in ESCC progression. Differentially expressed lncRNAs in ESCC were predicted by bioinformatics analysis. ZNF667-AS1, microRNA-1290 (miR-1290), and prune homolog 2 with BCH domain (PRUNE2) expression was determined with their relationship in cell biological processes analyzed also by means of gain- and loss-of-function assays. Xenograft mouse models were performed to re-produce the in vitro findings. We found a decline in ZNF667-AS1 expression in ESCC tissues and cell lines. ZNF667-AS1 overexpression indicated a favorable prognosis of ESCC sufferers. ZNF667-AS1 overexpression suppressed ESCC cell malignant potentials. ZNF667-AS1 reduced miR-1290 to result in upregulation of the miR-1290 target gene PRUNE2. The inhibiting property of ZNF667-AS1 on the malignant characteristics of ESCC cells was achieved by disrupting the miR-1290-mediated downregulation of PRUNE2. ZNF667-AS1 suppressed the tumorigenesis of ESCC in vivo. Collectively, our study demonstrates that ZNF667-AS1 can function as a tumor suppressor in ESCC by upregulating PRUNE2 and downregulating miR-1290.
NEAT1/hsa-miR-372–3p axis participates in rapamycin-induced lipid metabolic disorder
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Hence, this study provides unequivocal evidence that the down-regulation of hsa-miR-372–3p interferes with rapamycin-induced disruption of lipid homeostasis. Besides the metabolic regulation of hsa-miR-372–3p reported in this study, low hsa-miR-372–3p expression has also been reported to be associated with poor prognosis and tumor metastasis in HCC [42,43]. This means that despite the advantages of rapamycin in treating HCC, it still has the risk of promoting HCC through down-regulating hsa-miR-372–3p.
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Thousands of long non-coding RNAs (lncRNAs) have been discovered in human genomes by gene chip, next-generation sequencing, and/or other methods in recent years, which represent a significant subset of the universal genes involved in a wide range of biological functions. An abnormal expression of lncRNAs is associated with the growth, invasion, and metastasis of various types of human cancers, including hepatocellular carcinoma (HCC), which is an aggressive, highly malignant, and invasive tumor, and a poor prognosis in China. With a more in-depth understanding of lncRNA research for HCC and the emergence of new molecular-targeted therapies, the diagnosis, treatment, and prognosis of HCC will be considerably improved. Therefore, this review is expected to provide recommendations and directions for future lncRNA research for HCC.
LncRNA TMPO-AS1 Aggravates the Development of Hepatocellular Carcinoma via miR-429/GOT1 Axis; TMPO-AS1 aggravates the development of hepatocellular carcinoma
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MicroRNAs (miRNAs), a type of noncoding RNAs with a length of 20-24 nucleotides, play pivotal roles in regulating the expression of target genes at a post-transcriptional level.19,20 It has been widely recognized that lncRNAs serve as competing endogenous RNAs (ceRNAs) by competitively sponging miRNAs and then regulating specific cancers.21–23 TMPO-AS1 has been reported to regulate the progression of cervical cancer by sponging miR-577.12
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View full text

lncRNA OSER1-AS1 acts as a ceRNA to promote tumorigenesis in hepatocellular carcinoma by regulating miR-372-3p/Rab23 axis

Highlights

Abstract

Introduction

Section snippets

Clinical samples

OSER1-AS1 is highly expressed in HCC

Discussion

Acknowledgements

Cancer Cell

Biochem. Biophys. Res. Commun.

J. Urol.

Cell

Cancer statistics in China, 2015

CA A Cancer J. Clin.

Prophylactic liver transplantation for high-risk recurrent hepatocellular carcinoma

World J. Hepatol.

Recent developments with immunotherapy for hepatocellular carcinoma

Expert Opin. Biol. Ther.

MicroRNA binding sites in the coding region of mRNAs: extending the repertoire of post-transcriptional gene regulation

Bioessays

A SOX9-AS1/miR-5590-3p/SOX9 positive feedback loop drives tumor growth and metastasis in hepatocellular carcinoma through the Wnt/beta-catenin pathway

Mol. Oncol.

GEPIA: a web server for cancer and normal gene expression profiling and interactive analyses

Nucleic Acids Res.

Imprinted H19 oncofetal RNA is a candidate tumour marker for hepatocellular carcinoma

Mol. Pathol.

Epigenetic activation of the MiR-200 family contributes to H19-mediated metastasis suppression in hepatocellular carcinoma

Carcinogenesis