Multiple microRNAs as biomarkers for early breast cancer diagnosis

Jang,Ji Young; Kim,Yeon Soo; Kang,Kyung Nam; Kim,Kyo Hyun; Park,Yu Jin; Kim,Chul Woo

doi:10.3892/mco.2020.2193

Multiple microRNAs as biomarkers for early breast cancer diagnosis

Authors:
- Ji Young Jang
- Yeon Soo Kim
- Kyung Nam Kang
- Kyo Hyun Kim
- Yu Jin Park
- Chul Woo Kim
View Affiliations

Affiliations: BIOINFRA Life Science Inc., Jongno‑gu, Seoul 03127, Republic of Korea, DIOGENE Inc., Bundang‑gu, Seongnam‑Si 13486, Republic of Korea
Published online on: December 17, 2020 https://doi.org/10.3892/mco.2020.2193
Article Number: 31
Copyright: © Jang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

MicroRNA (miRNA or miR) is stably present in plasma. It has been reported that miRNA could be used for detecting cancer. Circulating miRNAs are being increasingly recognized as powerful biomarkers in a number of different pathologies, including in breast cancer. The aim of the current study was to establish and validate miRNA sets that are useful for the early diagnosis of breast cancer. Specifically, the current study intended to determine whether miRNA biomarkers were tumor‑specific and to statistically verify whether circulating miRNA analysis could be used for breast cancer diagnosis. In the present study, a total of nine candidate miRNA biomarkers were selected by examining reference miRNAs associated with the generation and progression of breast cancer to identify novel miRNAs that could be used to detect early breast cancer. A total of 226 plasma samples from patients with breast cancer were used. In addition, 146 plasma healthy samples were used as non‑cancer controls. These samples were divided into training and validation cohorts. The training cohort was used to identify a combination of miRNA that could detect breast cancer. The validation cohort was used to validate this combination of miRNA. Total RNAs were isolated from collected samples. A total of 9 miRNAs were quantified using reverse‑transcription quantitative PCR. A total of nine candidate miRNA expression levels were compared between patients with breast cancer and healthy controls. It was indicated that combinations of two or more of the nine miRNAs could detect breast cancer with higher accuracy than the use of a single biomarker. As a representative example, combinations of four miRNAs (miR‑1246+miR‑206+miR‑24+miR‑373) of the nine miRNAs had a sensitivity of 98%, a specificity of 96% and an accuracy of 97% for breast cancer detection in the validation cohort. The results of the present study suggest that multiple miRNAs could be used as potential biomarkers for early diagnosis of breast cancer. These biomarkers are expected to overcome limitations of mammography when used as an auxiliary diagnosis of mammography.

View Figures

View References

1	Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D and Bray F: Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 136:E359–E386. 2015.PubMed/NCBI View Article : Google Scholar
2	Wang L: Early diagnosis of breast cancer. Sensors. 17(1572)2017.PubMed/NCBI View Article : Google Scholar
3	Korean Breast Cancer Society. Early screening of breast cancer in Korea. J Korean Breast Cancer Soc. 5:225–234. 2002.
4	Devolli-Disha E, Manxhuka-Kërliu S, Ymeri H and Kutllovci A: Comparative accuracy of mammography and ultrasound in women with breast symptoms according to age and breast density. Bosn J Basic Med Sci. 9:131–136. 2009.PubMed/NCBI View Article : Google Scholar
5	Marrugo-Ramírez J, Mir M and Samitier J: Blood-Based cancer biomarkers in liquid biopsy: A promising non-invasive alternative to tissue biopsy. Int J Mol Sci. 19(2877)2018.PubMed/NCBI View Article : Google Scholar
6	Bai Y and Zhao H: Liquid biopsy in tumors: Opportunities and challenges. Ann Transl Med. 6(S89)2018.PubMed/NCBI View Article : Google Scholar
7	Castro-Giner F, Gkountela S, Donato C, Alborelli I, Quagliata L, Ng CKY, Piscuoglio S and Aceto N: Cancer diagnosis using a liquid biopsy: Challenges and expectations. Diagnostics (Basel). 8(31)2018.PubMed/NCBI View Article : Google Scholar
8	Simpson RJ, Lim JW, Moritz RL and Mathivanan S: Exosomes: Proteomic insights and diagnostic potential. Expert Rev Proteomics. 6:267–283. 2009.PubMed/NCBI View Article : Google Scholar
9	Chen X, Liang H, Zhang J, Zen K and Zhang CY: Horizontal transfer of microRNAs: Molecular mechanisms and clinical applications. Protein Cell. 3:28–37. 2012.PubMed/NCBI View Article : Google Scholar
10	Sanz-Rubio D, Martin-Burriel I, Gil A, Cubero P, Forner M, Khalyfa A and Marin JM: Stability of circulating Exosomal miRNAs in healthy subjects. Sci Rep. 8(10306)2018.PubMed/NCBI View Article : Google Scholar
11	Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, Pogosova-Agadjanyan EL, Peterson A, Noteboom J, O'Briant KC, Allen A, et al: Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci USA. 105:10513–10518. 2008.PubMed/NCBI View Article : Google Scholar
12	Salehi M and Sharifi M: Exosomal miRNAs as novel cancer biomarkers: Challenges and opportunities. J Cell Physiol. 233:6370–6380. 2018.PubMed/NCBI View Article : Google Scholar
13	Calin GA and Croce CM: MicroRNA signatures in human cancers. Nat Rev Cancer. 6:857–866. 2006.PubMed/NCBI View Article : Google Scholar
14	He L, He X, Lim LP, de Stanchina E, Xuan Z, Liang Y, Xue W, Zender L, Magnus J, Ridzon D, et al: A microRNA component of the p53 tumour suppressor network. Nature. 447:1130–1134. 2007.PubMed/NCBI View Article : Google Scholar
15	Lu J, Getz G, Miska EA, Alvarez-Saavedra E, Lamb J, Peck D, Sweet-Cordero A, Ebert BL, Mak RH, Ferrando AA, et al: MicroRNA expression profiles classify human cancers. Nature. 435:834–838. 2005.PubMed/NCBI View Article : Google Scholar
16	Calin GA, Ferracin M, Cimmino A, Di Leva G, Shimizu M, Wojcik SE, Iorio MV, Visone R, Sever NI, Fabbri M, et al: A MicroRNA signature associated with prognosis and progression in chronic lymphocytic leukemia. N Engl J Med. 353:1793–1801. 2005.PubMed/NCBI View Article : Google Scholar
17	Schetter AJ, Leung SY, Sohn JJ, Zanetti KA, Bowman ED, Yanaihara N, Yuen ST, Chan TL, Kwong DL, Au GK, et al: MicroRNA expression profiles associated with prognosis and therapeutic outcome in colon adenocarcinoma. JAMA. 299:425–436. 2008.PubMed/NCBI View Article : Google Scholar
18	Heneghan HM, Miller N, Lowery AJ, Sweeney KJ, Newell J and Kerin MJ: Circulating microRNAs as novel minimally invasive biomarkers for breast cancer. Ann Surg. 251:499–505. 2010.PubMed/NCBI View Article : Google Scholar
19	Zhao H, Shen J, Medico L, Wang D, Ambrosone CB and Liu S: A pilot study of circulating miRNAs as potential biomarkers of early stage breast cancer. PLoS One. 5(e13735)2010.PubMed/NCBI View Article : Google Scholar
20	Kim VN, Han J and Siomi MC: Biogenesis of small RNAs in animals. Nat Rev Mol Cell Biol. 10:126–139. 2009.PubMed/NCBI View Article : Google Scholar
21	Han J, Lee Y, Yeom KH, Kim YK, Jin H and Kim VN: The Drosha-DGCR8 complex in primary microRNA processing. Genes Dev. 18:3016–3027. 2004.PubMed/NCBI View Article : Google Scholar
22	Lee Y, Ahn C, Han J, Choi H, Kim J, Yim J, Lee J, Provost P, Rådmark O, Kim S and Kim VN: The nuclear RNase III Drosha initiates microRNA processing. Nature. 425:415–419. 2003.PubMed/NCBI View Article : Google Scholar
23	Esquela-Kerscher A and Slack FJ: Oncomirs-microRNAs with a role in cancer. Nat Rev Cancer. 6:259–269. 2006.PubMed/NCBI View Article : Google Scholar
24	Suzuki HI and Miyazono K: Emerging complexity of microRNA generation cascades. J Biochem. 149:15–25. 2011.PubMed/NCBI View Article : Google Scholar
25	Salido-Guadarrama I, Romero-Cordoba S, Peralta-Zaragoza O, Hidalgo-Miranda A and Rodríguez-Dorantes M: MicroRNAs transported by exosomes in body fluids as mediators of intercellular communication in cancer. Onco Targets Ther. 7:1327–1338. 2014.PubMed/NCBI View Article : Google Scholar
26	Lim SS and Kim YS: WO2016137031-Dumbell-structure oligonucleotide, nucleic acid amplification primer comprising same, and nucleic acid amplification method using same. KR Patent WO/2016/137031. DIOGENE Inc. Filed February 25, 2015; issued September 1, 2016.
27	Pinatel EM, Orso F, Penna E, Cimino D, Elia AR, Circosta P, Dentelli P, Brizzi MF, Provero P and Taverna D: miR-223 is a coordinator of breast cancer progression as revealed by bioinformatics predictions. PLoS One. 9(e84859)2014.PubMed/NCBI View Article : Google Scholar
28	Yoshikawa M, Iinuma H, Umemoto Y, Yanagisawa T, Matsumoto A and Jinno H: Exosome-encapsulated microRNA-223-3p as a minimally invasive biomarker for the early detection of invasive breast cancer. Oncol Lett. 15:9584–9592. 2018.PubMed/NCBI View Article : Google Scholar
29	Fu L, Li Z, Zhu J, Wang P, Fan G, Dai Y, Zheng Z and Liu Y: Serum expression levels of microRNA-382-3p, -598-3p, -1246 and -184 in breast cancer patients. Oncol Lett. 12:269–274. 2016.PubMed/NCBI View Article : Google Scholar
30	Li XJ, Ren ZJ, Tang JH and Yu Q: Exosomal MicroRNA miR-1246 Promotes Cell Proliferation, Invasion and Drug Resistance by Targeting CCNG2 in Breast Cancer. Cell Physiol Biochem. 44:1741–1748. 2017.PubMed/NCBI View Article : Google Scholar
31	Zhou Y, Wang M, Tong Y, Liu X, Zhang L, Dong D, Shao J and Zhou Y: miR-206 promotes cancer progression by targeting Full-Length Neurokinin-1 Receptor in breast cancer. Technol Cancer Res Treat. 18(1533033819875168)2019.PubMed/NCBI View Article : Google Scholar
32	Khodadadi-Jamayran A, Akgol-Oksuz B, Afanasyeva Y, Heguy A, Thompson M, Ray K, Giro-Perafita A, Sánchez I, Wu X, Tripathy D, et al: Prognostic role of elevated mir-24-3p in breast cancer and its association with the metastatic process. Oncotarget. 9:12868–12878. 2018.PubMed/NCBI View Article : Google Scholar
33	Lu K, Wang J, Song Y, Zhao S, Liu H, Tang D, Pan B, Zhao H and Zhang Q: miRNA-24-3p promotes cell proliferation and inhibits apoptosis in human breast cancer by targeting p27^Kip1. Oncol Rep. 34:995–1002. 2015.PubMed/NCBI View Article : Google Scholar
34	Eichelser C, Flesch-Janys D, Chang-Claude J, Pantel K and Schwarzenbach H: Deregulated Serum Concentrations of Circulating Cell-Free MicroRNAs miR-17, miR-34a, miR-155 and miR-373 in Human Breast Cancer Development and Progression. Clin Chem. 59:1489–1496. 2013.PubMed/NCBI View Article : Google Scholar
35	Piasecka D, Braun M, Kordek R, Sadej R and Romanska H: MicroRNAs in regulation of triple-negative breast cancer progression. J Cancer Res Clin Oncol. 144:1401–1411. 2018.PubMed/NCBI View Article : Google Scholar
36	Asaga S, Kuo C, Nguyen T, Terpenning M, Giuliano AE and Hoon DS: Direct serum assay for microRNA-21 concentrations in early and advanced breast cancer. Clin Chem. 57:84–91. 2011.PubMed/NCBI View Article : Google Scholar
37	Corcoran C, Friel AM, Duffy MJ, Crown J and O'Driscoll L: Intracellular and Extracellular MicroRNAs in Breast Cancer. Clin Chem. 57:18–32. 2011.PubMed/NCBI View Article : Google Scholar
38	Shimomura A, Shiino S, Kawauchi J, Takizawa S, Sakamoto H, Matsuzaki J, Ono M, Takeshita F, Niida S, Shimizu C, et al: Novel combination of serum microRNA for detecting breast cancer in the early stage. Cancer Sci. 107:326–334. 2016.PubMed/NCBI View Article : Google Scholar
39	Schrauder MG, Strick R, Schulz-Wendtland R, Strissel PL, Kahmann L, Loehberg CR, Lux MP, Jud SM, Hartmann A, Hein A, et al: Circulating micro-RNAs as potential blood-based markers for early stage breast cancer detection. PLoS One. 7(e29770)2012.PubMed/NCBI View Article : Google Scholar
40	Zhao C, Li X, Han B, You Z, Qu L, Liu C, Song J, Lian L and Yang N: Gga-miR-219b targeting BCL11B suppresses proliferation, migration and invasion of Marek's disease tumor cell MSB1. Sci Rep. 7(4247)2017.PubMed/NCBI View Article : Google Scholar
41	Løberg M, Lousdal ML, Bretthauer M and Kalager M: Benefits and harms of mammography screening. Breast Cancer Res. 17(63)2015.PubMed/NCBI View Article : Google Scholar
42	Guo R, Lu G, Qin B and Fei B: Ultrasound imaging technologies for breast cancer detection and management. Ultrasound Med Biol. 44:37–70. 2018.PubMed/NCBI View Article : Google Scholar
43	Lebron-Zapata L and Jochelson MS: Overview of breast cancer screening and diagnosis. PET Clin. 13:301–323. 2018.PubMed/NCBI View Article : Google Scholar
44	Alimirzaie S, Bagherzadeh M and Akbari MR: Liquid biopsy in breast cancer: A comprehensive review. Clin Genet. 95:643–660. 2019.PubMed/NCBI View Article : Google Scholar
45	Hamam R, Hamam D, Alsaleh KA, Kassem M, Zaher W, Alfayez M, Aldahmash A and Alajez NM: Circulating microRNAs in breast cancer: Novel diagnostic and prognostic biomarkers. Cell Death Dis. 8(e3045)2017.PubMed/NCBI View Article : Google Scholar
46	He Y, Deng F, Yang S, Wang D, Chen X, Zhong S, Zhao J and Tang J: Exosomal microRNA: A novel biomarker for breast cancer. Biomark Med. 12:177–188. 2018.PubMed/NCBI View Article : Google Scholar
47	Hannafon BN, Trigoso YD, Calloway CL, Zhao YD, Lum DH, Welm AL, Zhao ZJ, Blick KE, Dooley WC and Ding WQ: Plasma exosome microRNAs are indicative of breast cancer. Breast Cancer Res. 18(90)2016.PubMed/NCBI View Article : Google Scholar