Circulating miRNA-21 and miRNA-23a Expression Signature as Potential Biomarkers for Early Detection of Non-Small-Cell Lung Cancer

Helal   Fouad    Hetta; Asmaa   Mohammad   Zahran; Engy   A.   Shafik; Reham   I.    El-Mahdy; Nahed   A.   Mohamed; Emad   Eldin   Nabil; Hend    M.   Esmaeel; Ola   A.   Alkady; Azza      Elkady; Dina   A.   Mohareb; Amal      Hosni; Mohammed   Mahmoud    Mostafa; Abeer      Elkady

Abstract

Background and Aim: Lung Cancer (LC) is a major cancer killer worldwide, and 5-yr survival is extremely poor (≤15%), accentuating the need for more effective diagnostic and therapeutic strategies. Studies have shown cell-free microRNAs (miRNAs) circulating in the serum and plasma with specific expression in cancer, indicating the potential of using miRNAs as biomarkers for cancer diagnosis and therapy. This study aimed to identify differentially-expressed two miRNAs in the plasma of Non-Small Cell Lung Cancer (NSCLC) patients that might be a clinically useful tool for lung cancer early detection. miRNA-21 is one of the most abundant oncomirs. miRNA-23a functions as an oncogene in several human cancers, however, its clinical value has not been investigated in NSCLC.

Materials and Methods: A case-control study was conducted in Assiut University Hospital, Egypt, from 2017 to 2018. Plasma samples were obtained from 45 NSCLC patients. The expression level of miR-21 and miRNA-23a was detected by qRT-PCR and compared to 40 healthy control subjects. The relation between both miRNAs and clinicopathological parameters was evaluated.

Results: The expression level of miR-21 and miRNA-23a was significantly up-regulated (36.9 ± 18.7 vs. 1.12 ± 0.84 and 24.7 ± 19.09 vs. 1.16 ± 0.45) in NSCLC compared to matched controls (P<0.0001each). There was a significant difference in the level of plasma miRNA-21 and miRNA- 23a expression between the different grades of the disease (P = 0.032 and P = 0.001, respectively). The plasma miRNA-21 and miRNA-23a levels in the lung cancer patients with distant metastasis (n = 20) were significantly higher than those in the patients without metastasis (n = 25) (P<0.0001 each), the expression of miR-21 and miRNA-23a was significantly associated with tumor size (P = 0.001, P = 0.0001, respectively), but not significantly related to lymph node metastasis (P = 0.687 and 0.696, respectively). A positive correlation was observed between miRNA-21 and miRNA-23a (r = 0.784, P<0.01), There was no significant difference in the plasma miRNA-21 and miRNA-23a levels in the lung cancer patients with different histopathological types.

Conclusion: miR-21 and miR-23a might play an oncogenic role in LC and is a poor prognostic factor. Switching off miRNA-21 and miRNA-23a may improve the treatment of LC. Our results must be verified by large-scale prospective studies with standardized methodology.

Keywords: Biomarkers, early detection, miR-21, miR-23a, non-small-cell lung cancer, tumor.

Graphical Abstract

[1] 
Torre L, Bray F, Siegel R, et al. Global cancer statistics, 2012. CA Cancer J Clin  2015; 65: 87-108.
[2] 
Sittka A, Schmeck B. MicroRNAs in the lung In microRNA cancer regulationSpringer.  Heidelberg 2013; pp. 121-34.
[3] 
Rudin CM, Durinck S, Stawiski EW, et al. Comprehensive genomic analysis identifies SOX2 as a frequently amplified gene in small-cell lung cancer. Nat Genet  2012; 44: 1111-6.
[4] 
Travis WD, Brambilla E, Burke AP, et al. Introduction to the 2015 World Health Organization classification of tumors of the lung, pleura, thymus, and heart. J Thorac Oncol  2015; 10: 1240-2.
[5] 
George J, Lim JS, Jang SJ, et al. Comprehensive genomic profiles of small cell lung cancer. Nature  2015; 524: 47.
[6] 
Lind H, Zienolddiny S, Ryberg D, et al. Interleukin 1 receptor antagonist gene polymorphism and risk of lung cancer: a possible interaction with polymorphisms in the interleukin 1 beta gene. Lung Cancer  2005; 50: 285-90.
[7] 
Sun S, Schiller JH, Gazdar AF. Lung cancer in never smokers-a different disease. Nat Rev Cancer  2007; 7: 778.
[8] 
Amos C, Xu W, Spitz M. Is there a genetic basis for lung cancer susceptibility? In chemoprevention of cancer.  Springer Chicago 1999; pp. 3-12.
[9] 
Bach PB, Kelley MJ, Tate RC, et al. Screening for lung cancer: a review of the current literature. Chest  2003; 123: S72-82.
[10] 
Heuvelmans MA, Groen HJ, Oudkerk M. Early lung cancer detection by low-dose CT screening: therapeutic implications. Expert Rev Respir Med  2017; 11: 89-100.
[11] 
Lee RC, Feinbaum RL, Ambros V. The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell  1993; 75: 843-54.
[12] 
Pasquinelli AE, Reinhart BJ, Slack F, et al. Conservation of the sequence and temporal expression of let-7 heterochronic regulatory RNA. Nature  2000; 408: 86.
[13] 
Johnson CD, Esquela-Kerscher A, Stefani G, et al. The let-7 microRNA represses cell proliferation pathways in human cells. Cancer Res  2007; 67: 7713-22.
[14] 
Wang L, Zhang L-F, Wu J, et al. IL-1β-mediated repression of microRNA-101 is crucial for inflammation-promoted lung tumorigenesis. Cancer Res  2014; 74(17): 4720-30.
[15] 
Gasparini P, Cascione L, Landi L, et al. microRNA classifiers are powerful diagnostic/prognostic tools in ALK-, EGFR-, and KRAS-driven lung cancers. Proc Natl Acad Sci USA  2015; 112: 14924-9.
[16] 
Hu L, Ai J, Long H, et al. Integrative microRNA and gene profiling data analysis reveals novel biomarkers and mechanisms for lung cancer. Oncotarget  2016; 7: 8441.
[17] 
Mitchell PS, Parkin RK, Kroh EM, et al. Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci USA  2008; 105: 10513-8.
[18] 
Qu WQ, Liu L, Yu Z. Clinical value of microRNA-23a upregulation in non-small cell lung cancer. Int Clin Exp Med  2015; 8: 13598-603.
[19] 
Capodanno A, Boldrini L, Proietti A, et al. Let-7g and miR-21 expression in non-small cell lung cancer: correlation with clinicopathological and molecular features. Int J Oncol  2013; 43: 765-74.
[20] 
Gao Y, Dai M, Liu H, et al. Diagnostic value of circulating miR-21: an update meta-analysis in various cancers and validation in endometrial cancer. Oncotarget  2016; 7: 68894-908.
[21] 
Jazbutyte V, Thum T. MicroRNA-21: from cancer to cardiovascular disease. Curr Drug Targets  2010; 11: 926-35.
[22] 
Meng F, Henson R, Wehbe–Janek H, et al. MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer. Gastroenterology  2007; 133: 647-58.
[23] 
Frankel LB, Christoffersen NR, Jacobsen A, et al. Programmed cell death 4 (PDCD4) is an important functional target of the microRNA miR-21 in breast cancer cells. J Biol Chem  2008; 283(2): 1026-33.
[24] 
Asangani IA, Rasheed SA, Nikolova D, et al. MicroRNA-21 (miR-21) post-transcriptionally downregulates tumor suppressor PDCD4 and stimulates invasion, intravasation and metastasis in colorectal cancer. Oncogene  2008; 27: 2128.
[25] 
Liu S, Sun X, Wang M, et al. A microRNA 221- and 222-mediated feedback loop maintains constitutive activation of NFkappaB and STAT3 in colorectal cancer cells. Gastroenterology  2014; 147: 847-59.
[26] 
Ma G, Dai W, Sang A, et al. Upregulation of microRNA-23a/b promotes tumor progression and confers poor prognosis in patients with gastric cancer. Int Clin Exp Pathol  2014; 7: 8833.
[27] 
Zhang X-W, Liu N, Chen S, et al. High microRNA-23a expression in laryngeal squamous cell carcinoma is associated with poor patient prognosis. Diagn Pathol  2015; 10: 22.
[28] 
Cao M, Seike M, Soeno C, et al. MiR-23a regulates TGF-β-induced epithelial-mesenchymal transition by targeting E-cadherin in lung cancer cells. Int Oncol  2012; 41: 869-75.
[29] 
Molina JR, Yang P, Cassivi SD, et al. Non-small cell lung cancer: epidemiology, risk factors, treatment, and survivorship  Mayo Clin proc Mayo Clinic 2008; 83: 584-94.
[30] 
Hamam R, Ali AM, Alsaleh KA, et al. microRNA expression profiling on individual breast cancer patients identifies novel panel of circulating microRNA for early detection. Sci Rep  2016; 6: 25997.
[31] 
Petricoin EF 3rd, Ornstein DK, Paweletz CP, et al. Serum proteomic patterns for detection of prostate cancer. J Natl Cancer Inst  2002; 94: 1576-8.
[32] 
MacFarlane LA. R Murphy P. MicroRNA: biogenesis, function and role in cancer. Curr Genomics  2010; 11: 537-61.
[33] 
Zhou X, Yang P-C. MicroRNA: a small molecule with a big biological impact. MicroRNA  2012; 1(1): 1.
[34] 
Shen J, Liu Z, Todd NW, et al. Diagnosis of lung cancer in individuals with solitary pulmonary nodules by plasma microRNA biomarkers. BMC Cancer  2011; 11: 374.
[35] 
Shen J, Todd NW, Zhang H, et al. Plasma microRNAs as potential biomarkers for non-small-cell lung cancer. Lab Invest  2011; 91: 579.
[36] 
Markou A, Sourvinou I, Vorkas P, et al. Clinical evaluation of microRNA expression profiling in non small cell lung cancer. Lung Cancer  2013; 81: 388-96.
[37] 
Tang D, Shen Y, Wang M, et al. Identification of plasma microRNAs as novel noninvasive biomarkers for early detection of lung cancer. Eur J Cancer Prev  2013; 22: 540-8.
[38] 
Markou A, Zavridou M, Lianidou ES. miRNA-21 as a novel therapeutic target in lung cancer. Lung Cancer: Targets Ther   2016; 7: 19.
[39] 
Novello S, Barlesi F, Califano R, et al. Metastatic non-small-cell lung cancer: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol  2016; 27: v1-v27.
[40] 
Vucic EA, Thu KL, Pikor LA, et al. Smoking status impacts microRNA mediated prognosis and lung adenocarcinoma biology. BMC Cancer  2014; 14: 778.
[41] 
Izzotti A, Longobardi M, La Maestra S, et al. Release of microRNAs into body fluids from ten organs of mice exposed to cigarette smoke. Theranostics  2018; 8: 2147.
[42] 
Suzuki K, Yamada H, Nagura A, et al. Association of cigarette smoking with serum microRNA expression among middle-aged Japanese adults. Fujian Med J  2016; 2: 1-5.
[43] 
Qu W-Q, Liu L, Yu Z. Clinical value of microRNA-23a upregulation in non-small cell lung cancer. Int Clin Exp Med  2015; 8: 13598.
[44] 
Liu X-G, Zhu W-Y, Huang Y-Y, et al. High expression of serum miR-21 and tumor miR-200c associated with poor prognosis in patients with lung cancer. Med Oncol  2012; 29: 618-26.
[45] 
Wei J, Gao W, Zhu C-J, et al. Identification of plasma microRNA-21 as a biomarker for early detection and chemosensitivity of non-small cell lung cancer. Chin J Cancer  2011; 30: 407.

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