Differential Expression of miR-101 and miR-744 in Nasopharyngeal Carcinoma in Pahang State of Malaysia

Azmir AHMAD; Haziq Abdul WAHID; Wan Ishlah LEMAN; Noor Syamila OTHMAN; Azlina Abdul RAHMAN; Kahairi ABDULLAH; Siti Aesah Naznin MUHAMMAD; Mohd. Arifin KADERI

doi:10.48048/wjst.2019.6285

Authors

Azmir AHMAD Department of Biomedical Science, Kulliyyah of Allied Health Sciences, International Islamic University Malaysia, Jalan Sultan Haji Ahmad Shah, Bandar Indera Mahkota, Pahang
Haziq Abdul WAHID Department of Biomedical Science, Kulliyyah of Allied Health Sciences, International Islamic University Malaysia, Jalan Sultan Haji Ahmad Shah, Bandar Indera Mahkota, Pahang
Wan Ishlah LEMAN Department of Otorhinolaryngology-Head and Neck Surgery, Kulliyyah of Medicine, International Islamic University Malaysia, Jalan Sultan Haji Ahmad Shah, Bandar Indera Mahkota, Pahang
Noor Syamila OTHMAN Department of Biomedical Science, Kulliyyah of Allied Health Sciences, International Islamic University Malaysia, Jalan Sultan Haji Ahmad Shah, Bandar Indera Mahkota, Pahang
Azlina Abdul RAHMAN Department of Pathology, Hospital Tengku Ampuan Afzan, Jalan Tanah Putih, Pahang
Kahairi ABDULLAH Department of Otorhinolaryngology-Head and Neck Surgery, Kulliyyah of Medicine, International Islamic University Malaysia, Jalan Sultan Haji Ahmad Shah, Bandar Indera Mahkota, Pahang
Siti Aesah Naznin MUHAMMAD Department of Pathology and Laboratory Medicine, Kulliyyah of Medicine, International Islamic University Malaysia, Jalan Sultan Haji Ahmad Shah, Bandar Indera Mahkota, Pahang
Mohd. Arifin KADERI Department of Biomedical Science, Kulliyyah of Allied Health Sciences, International Islamic University Malaysia, Jalan Sultan Haji Ahmad Shah, Bandar Indera Mahkota, Pahang

DOI:

https://doi.org/10.48048/wjst.2019.6285

Keywords:

Nasopharyngeal carcinoma, miR-101, miR-744, differential expression, diagnostic biomarker

Abstract

Previous study found that microRNA-101 (miR-101) and microRNA-744 (miR-744) were deregulated in head and neck cancers and were implicated in nasopharyngeal carcinoma (NPC) carcinogenesis. Thus, this study aimed to determine the expression of miR-101 and miR-744 in NPC and analyse the utility of these microRNAs (miRNAs) as diagnostic biomarkers. Total RNA was extracted from 31 NPC and 7 non-NPC control formalin-fixed paraffin-embedded (FFPE) samples. Complementary DNA (cDNA) was synthesized from the total RNA and proceeded with quantitative real-time polymerase chain reaction. Differential expression of miR-101 and miR-744 were calculated from quantification cycle (Cq) data using 2^-ΔΔCq calculation. The performance of these miRNAs were calculated using receiver operating characteristic (ROC) curve analysis. The differential expression for miR-101 and miR-744 were -1.39 (p < 0.05) and 2.48 (p > 0.05), respectively, where the deregulations were consistent with the previous report. The area under curve for miR-101, miR-744 and combination of miR-101 and miR-744 were 0.654 (95 % CI: 0.465 - 0.844), 0.588 (95 % CI: 0.368 - 0.808) and 0.626 (95 % CI: 0.481 - 0.771), respectively. However, re-analysis using balanced sample size between NPC and non-NPC control group showed the value decreased to 0.653 (95 % CI: 0.347 - 0.959) for miR-101 but increased to 0.827 (95 % CI: 0.601 - 1.000) for miR-744 and 0.758 (95 % CI: 0.576 - 0.939) for the combination of miR-101 and miR-744, indicating the importance of having a balanced sample size. We have successfully determined the expression of miR-101 and miR-744 in NPC samples. We also demonstrated statistically the utility of these miRNAs as diagnostic biomarkers.

Downloads

Download data is not yet available.

Metrics

Metrics Loading ...

References

M Azizah Ab, IT Nor Saleha, A Noor Hashimah, ZA, Azmah and W Mastulu. Malaysian National Cancer Registry Report 2007-2011. Ministry of Health, Malaysia. Available at: http://nci.moh.gov.my/index.php/en/announcement/340-malaysian-national-cancer-registry-report-2007-2011, accessed January 2018.

BCR Devi, P Pisani, TS Tang and DM Parkin. High incidence of nasopharyngeal cancer carcinoma in Native People of Sarawak, Borneo Island. Epidemiol. Biomarkers Prev. 2004; 13, 482-6.

GCC Lim, S Rampal and Y Halimah. Cancer Incidence in Peninsular Malaysia, 2003-2005. The Third Report of the National Cancer Registry Malaysia. Ministry of Health, Malaysia. Available at: http://www.moh.gov.my/images/gallery/Report/Cancer/CancerIncidenceinPeninsularMalaysia2003-2005x1x.pdf, accessed January 2018.

K Breving and A Esquela-Krescher. The complexities of microRNA regulation: Mirandering around the rules. Int. J. Biochem. Cell Biol. 2006; 42, 1316-29.

R Schickel, B Boyerinas, SM Park and ME Peter. MicroRNAs: Key players in the immune system, differentiation, tumorigenesis and cell death. Oncogene 2008; 27, 5959-74.

A Esquela-Krescher and FJ Slack. Oncomirs-microRNA with a role in cancer. Nat. Rev. Cancer 2006; 6, 259-69.

S Pfeffer and O Voinnet. Viruses, microRNAs and cancer. Oncogene 2006; 25, 6211-9.

SK Shenouda and SK Alahari. MicroRNA function in cancer: Oncogene or a tumor suppressor? Cancer Metastasis Rev. 2009; 28, 369-78.

T Spence, J Bruce, KW Yip and FF Liu. MicroRNAs in nasopharyngeal carcinoma. Chin. Clin. Oncol. 2016; 5, 17.

J Lu, H Luo, X Liu, Y Peng, B Zhang, L Wang, X Xu, X Peng, G Li, W Tian, ML He, H Kung and XP Li. miR-9 targets CXCR4 and functions as a potential tumor suppressor in nasopharyngeal carcinoma. Carcinogenesis 2014; 35, 554-63.

K Cai, Y Wan, G Sun, L Shi, X Bao and Z Wang. Let-7a inhibits proliferation and induces apoptosis by targeting EZH2 in nasopharyngeal carcinoma cells. Oncol. Rep. 2012; 28, 2101-6.

N Liu, LL Tang, Y Sun, RX Cui, HY Wang, BJ Huang, QM He, W Jiang and J Ma. MiR-29c suppresses invasion and metastasis by targeting TIAM1 in nasopharyngeal carcinoma. Cancer Lett. 2013; 329, 181-8.

JX Zhang, D Qian, FW Wang, DZ Liao, JH Wei, ZT Tong, J Fu, XX Huang, YJ Liao, HX Deng, YX Zeng, D Xie and SJ Mai. MicroRNA-29c enhances the sensitivities of human nasopharyngeal carcinoma to cisplatin-based chemotherapy and radiotherapy. Cancer Lett. 2013; 329, 91-8.

EY Choy, KL Siu, KH Kok, RW Lung, CM Tsang, KF To, DL Kwong, SW Tsao and DY Jin. An Epstein-Barr virus-encoded microRNA targets PUMA to promote host cell survival. J. Exp. Med. 2008; 205, 2551-60.

L Dölken, G Malterer, S Erhard, S Kothe, CC Friedel, G Suffert, L Marcinowski, N Motsch, S Barth, M Beitzinger, D Lieber, SM Bailer, R Hoffmann, Z Ruzsics, E Kremmer, S Pfeffer, R Zimmer, UH Koszinowski, F Grässer, G Meister and J Haas. Systematic analysis of viral and cellular microRNA targets in cells latently infected with human gamma-herpesviruses by RISC immunoprecipitation assay. Cell Host Microbe. 2010; 7, 324-34.

L Cai, Y Ye, Q Jiang, Y Chen, X Lyu, J Li, S Wang, T Liu, H Cai, K Yao, JL Li and X Li. Epstein-Barr virus-encoded microRNA BART1 induces tumour metastasis by regulating PTEN-dependent pathways in nasopharyngeal carcinoma. Nat. Commun. 2015; 6, 7353.

Z Sha, X Zu, N Li, Y Li and D Li. Proto-oncogenic miR-744 is upregulated by transcription factor c-Jun via promoter activation mechanism. Oncotarget 2016; 7, 64977-86.

XR Tang, X Wen, QM He, YQ Li, XY Ren, XJ Yang, J Zhang, YQ Wang, J Ma and N Liu. MicroRNA-101 inhibits invasion and angiogenesis through targeting ITGA3 and its systemic delivery inhibits lung metastasis in nasopharyngeal carcinoma. Cell Death Dis. 2017; 8, e2566.

S Chen, H Wang, WL Ng, WJ, Curran and Y Wang. Radiosensitizing effects of ectopic miR-101 on non-small-cell lung cancer cells depend on the endogenous miR-101 level. Int. J. Radiat. Oncol. Biol. Phys. 2011; 81, 1525-9.

NM Alajez, W Shi, ABY Hui, J Bruce, M Lenarduzzi, E Ito, S Yue, BO Sullivan and FF Liu. Enhancer of Zeste homolog 2 (EZH2) is overexpressed in recurrent nasopharyngeal carcinoma and is regulated by miR-26a, miR-101 and miR-98. Cell Death Dis. 2010; 1, e85.

Q Sun, T Liu, T Zhang, S Du, GX Xie, X Lin, L Chen and Y Yuan. MiR-101 sensitizes human nasopharyngeal carcinoma cells to radiation by targeting stathmin 1. Mol. Med. Rep. 2015; 11, 3330-6.

Y Fang, X Zhu, J Wang, N Li, D Li, N Sakib, Z Sha and W Song. MiR-744 functions as a proto-oncogene in nasopharyngeal carcinoma progression and metastasis via transcriptional control of ARHGAP5. Oncotarget 2015; 6, 13164-75.

Z Sha, X Zhu, N Li, Y Li and D Li. Proto-oncogenic miR-744 is up-regulated by transcription factor c-Jun via a promoter activation mechanism. Oncotarget 2016; 7, 64977-86.

Q Yu, F Zhang, Z Du and Y Xiang. Up-regulation of serum miR-744 predicts poor prognosis in patients with nasopharyngeal carcinoma. Int. J. Clin. Exp. Med. 2015; 8, 13296-302.

AM Nurul-Syakima, C Yoke-Kqueen, AR Sabariah, MS Shiran, A Singh and L Learn-Han. Differential microRNA expression and identification of putative microRNA targets and pathways in head and neck cancers. Int. J. Mol. Med. 2011; 28, 327-36.

T Li, JX Chen, XP Fu, S Yang, Z Zhang, KhH Chen and Y Li. microRNA expression profiling of nasopharyngeal carcinoma. Oncol. Rep. 2011; 25, 1353-63.

N Liu, NY Chen, RX Cui, WF Li, Y Li, RR Wei, MY Zhang, Y Sun, BJ Huang, M Chen, QN He, N Jiang, L Chen, WC Cho, JP Yun, J Zeng, LZ Liu, L Li, Y Guo, HY Wang and J Ma. Prognostic value of a microRNA signature in nasopharyngeal carcinoma: A microRNA expression analysis. Lancet Oncol. 2012; 13, 633-41.

N-SA Mutalib, L Learn-Han, SM Sidik, SA Rahman, ASM Singh. And C Yoke-Kqueen. miR-181a regulates multiple pathways in hypopharyngeal squamous cell carcinoma. Afr. J. Biotechnol. 2012; 11, 6129-37.

YK Cheah, RW Cheng, SK Yeap, CH Khoo and HS See. Analysis of TP53 gene expression and p53 level of human hypopharyngeal FaDu (HTB-43) head and neck cancer cell line after microRNA-181a inhibition. Genet. Mol. Res. 2014; 13, 1679-83.

AB Hui, W Shi, PC Boutros, N Miller, M Pintilie, T Fyles, D McCready, D Wong, K Gerster, L Waldron, I Jurisica, L Penn and FF Liu. Robust global micro-RNA profiling with formalin-fixed paraffin-embedded breast cancer tissues. Lab. Invest. 2009; 89, 597-606.

RS Goswami, L Waldron, J Machado, NK Cervigne, W Xu, PP Reis, DJ Bailey, L Jurisica, MR Crump and S Kamel-Reid. Optimization and analysis of a quantitative real-time PCR-based technique to determine microRNA expression in formalin-fixed paraffin-embedded samples. BMC Biotechnol. 2010; 10, 47.

S Romero-Cordoba, S Rodriguez-Cuevas, R Rebollar-Vega, V Quintanar-Jurado, A Maffuz-Aziz, G Jimenez-Sanchez, V Bautista-Piňa, R Arellano-Llamas and A Hidalgo-Miranda. Identification and pathway analysis of microRNAs with no previous involvement in breast cancer. PloS One 2012; 7, e31904.

Z Vojtechova, J Zavadil, J Klozar, M Grega and R Tachezy. Comparison of the miRNA expression profiles in fresh frozen and formalin-fixed paraffin-embedded tonsillar tumors. PloS One 2017; 12, e0179645.

A Torres, K Torres, P Wdowiak, T Paszkowski and R Maciejewski. Selection and validation of endogenous controls for microRNA expression studies in endometrioid endometrial cancer tissues. Gynecol. Oncol. 2013; 130, 588-94.

K Hajian-Tilaki. Receiver Operating Characteristic (ROC) curve analysis for medical diagnostic test evaluation. Caspian J. Intern. Med. 2013; 4, 627-35.

J Wang, JT Yu, L Tan, Y Tian, J Ma, CC Tan, HF Wang, Y Liu, MS Tan, T Jiang and L Tan. Genome-wide circulating microRNA expression profiling indicates biomarkers for epilepsy. Sci. Rep. 2015; 5, 9522.

MB Wozniak, G Scelo, DC Muller, A Mukeria, D Zaridze and P Brennan. Circulating microRNAs as non-invasive biomarkers for early detection of non-small-cell lung cancer. PloS One 2015; 10, e0125026.