Abstract
Abnormal interaction between non-coding RNAs has been demonstrated to be a common molecular event in various human cancers, but its significance and underlying mechanisms have not been well documented. RNA-binding proteins (RBPs) are key regulators of RNA transcription and post-transcriptional processing. In this study, we found that RNA-binding protein 24 (RBM24) was frequently downregulated in nasopharyngeal carcinoma (NPC). The restoration of RBM24 expression suppressed NPC cellular proliferation, migration and invasion and impeded metastatic colonization in mouse models. Microarray analyses revealed that miR-25 expression was upregulated by RBM24 expression in NPC cells. Similarly, ectopic miR-25 expression suppressed NPC cellular growth and motility by targeting the pro-oncogenic lncRNA MALAT1, and the knockdown of MALAT1 expression exhibited similar effects as RBM24 restoration in NPC cells. Overall, these findings suggest a novel role of RBM24 as a tumor suppressor. Mechanistically, RBM24 acts at least in part through upregulating the expression of miR-25, which in turn targets MALAT1 for degradation.
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Main
Nasopharyngeal carcinoma (NPC) is a highly malignant cancer that often invades adjacent regions and metastasizes to regional lymph nodes and distant organs. Although early-stage NPC is highly radiocurable, the treatment results of locoregionally advanced NPC have been disappointing.1, 2 Therefore, elucidation of the molecular mechanisms underlying the tumorigenicity, invasion and metastasis of NPC is very important for the treatment of this disease.
Recently, comprehensive microarray analysis has revealed a microRNA (miRNA) signature that is significantly associated with the prognosis and progression of NPC.3, 4, 5 Among the numerous differentially expressed miRNAs in NPC, three miRNAs, including miR-29c, miR-9 and miR-26a, have been shown to be significantly downregulated and have been extensively studied in association with this disease.5, 6, 7, 8, 9, 10, 11, 12 Accumulating evidence indicates that long noncoding RNAs (lncRNAs) are frequently deregulated in the malignant transformation and progression of various types of cancer, including NPC.13, 14, 15, 16, 53, 54 Briefly, 2.5 μg total RNA was labeled with pCp-DY647 (Dharmacon, Lafayette, CO, USA). After hybridization, the arrays were scanned with a LuxScan 10 K Microarray Scanner (CapitalBio, Bei**g, China), and the resulting images were analyzed with GenePix Pro 6.0 software (Axon Instruments, Foster City, CA, USA).
In vivo experiments
Female 4- to 5-week-old athymic mice were purchased (BALB/c nu/nu; Guangdong Medical Laboratory Animal Center, Guangzhou, China) and were maintained under a specific pathogen-free environment. All animal experiments were approved by the Institutional Animal Care and Use Committee of Sun Yat-sen University Cancer Center. For the tumor xenograft experiments, tumor cells (5 × 104 or 1 × 106 cells/tumor in 100 μl of serum-free culture medium) were suspended in 200 μl RPMI 1640 complete culture medium with 25% Matrigel (BD Biosciences, Bedford, MA, USA) and inoculated subcutaneously into the right flanks of the nude mice. The mice were monitored daily for palpable tumor formation, and tumors were measured using a Vernier caliper, weighed and photographed. Tumor width (W) and length (L) were measured every 2 days. RBM24 expression was repressed by the addition of doxycycline (1 g/l) to the drinking water until the mice were killed at 3 (5 × 104 cells/tumor, n=16) or 9 weeks (1 × 106 cells/tumor, n=22) after inoculation. Then, the tumors were isolated and weighed. Tumor volumes were calculated using the formula V=1/2 (L × W2).
Accession numbers
The Gene Expression Omnibus database accession number for the miRNA array data reported in this paper is GSE66878.
Statistical analysis
All in vitro experiments were repeated at least three times unless stated otherwise. Differences among the groups and treatments were determined by Student’s t-test unless stated otherwise. Kaplan–Meier survival analyses were performed to compare the survival times between the RBM24-induced and non-induced mice, and the log-rank test was used to generate P-values. The differences were considered significant at a P<0.05.
Accession codes
Abbreviations
- RBPs:
-
RNA-binding proteins
- RBM24:
-
RNA-binding protein 24
- RRM:
-
RNA recognition motif
- Dox:
-
Doxycycline
- WT:
-
Wild type
- Ago2:
-
Argonaute2
- lncRNAs:
-
long noncoding RNAs
- MALAT1:
-
metastasis associated in lung adenocarcinoma transcript 1
- NPC:
-
nasopharyngeal carcinoma
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Acknowledgements
This study was supported by the grants from the Ministry of Science and Technology of China (2012CB967003 and 2015AA020931) and the National Natural Science Foundation of China (91440106, 81230045 and 81202137).
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Hua, WF., Zhong, Q., **a, TL. et al. RBM24 suppresses cancer progression by upregulating miR-25 to target MALAT1 in nasopharyngeal carcinoma. Cell Death Dis 7, e2352 (2016). https://doi.org/10.1038/cddis.2016.252
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DOI: https://doi.org/10.1038/cddis.2016.252
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