Musa EKİCİ, Öner ODABAŞ, Yakup DÜLGEROĞLU, Gönül ERDEN, Ahmet YEŞİLKURT, Fatma UÇAR, Gülfer ÖZTÜRK

Malign ve benign prostat hastalıklarında miR-21 ve miR-34a serum düzeylerinin tanısal etkinliği

Amaç: Bu çalışmada, benign prostat hiperplazisi, kronik prostatit ve prostat kanseri ayrımında miR-21 ve miR-34a serum seviyelerinin tanısal etkinliğinin belirlenmesi amaçlanmıştır.Gereç ve Yöntemler: Prostat iğne biyopsisi yapılan toplamda 70 hastadan (25 benign prostat hiperplazisi, 10 kronik prostatit ve 35 prostat kanseri) kan örnekleri alındı. Uygun koşullarda serum eldesinden sonra RNA izolasyonu, cDNA sentezi ve qRT-PCR analizi Rotor-Gene® Q (Qiagen, Germany) cihazında Qiagen marka kitler kullanılarak gerçekleştirildi. Normalizasyon için referans gen olarak RNU6 kullanılarak -∆Ct değerleri hesaplandı. Tüm istatistiksel hesaplamalarda -∆Ct değerleri kullanıldı. Bulgular: Benign prostat hiperplazisine kıyasla kronik prostatit ve kanser gruplarında miR-21 serum seviyelerinin upregüle olduğu ve gruplar arasındaki farklılığın istatistiksel olarak anlamlı olduğu görülmüştür (sırasıyla p=0.021 ve p=0.001). miR-21’in tek başına spesifisite ve sensitivite değerleri benign prostat hiperplazisi ile prostat kanseri ayrımında %56 ve %86 olarak tespit edilmiştir. miR-21’in miR-34a ile kombinasyonunun spesifisite ve sensitivite değerleri benign prostat hiperplazisi ile prostat kanseri ayrımında %84 ve %71 olarak hesaplanmıştır.Sonuç: Bu çalışma ile miR-21 ve miR-21/miR-34a kombinasyonunun prostat kanseri tanısında biyobelirteç adayı olabilecek tanısal perfomansa sahip olduğu gösterilmiştir. Ayrıca, miR-21 seviyelerinde benign prostat hiperplazisine kıyasla kronik prostatit ve prostat kanserinde kademeli bir yüksekliğin olması, moleküler düzeyde gerçekleşen inflamasyon ve kanser dönüşümü süreçlerinin dolaşımdaki mikroRNA profiline de yansıdığını düşündürmektedir.

Anahtar Kelimeler:

Prostat kanseri, Prostatit, Benign prostat hiperplazisi, mikroRNA, Tanısal etkinlik

Diagnostic efficiency of miR-21 and miR-34a serum levels in malign and benign prostate diseases

Objective: In this study aimed to determine the diagnostic efficiency of miR-21 and miR-34a serum levels in the discrimination of benign prostatic hyperplasia, chronic prostatitis, and prostate cancer.Materials and Methods: Blood samples were taken from 70 patients (25 benign prostatic hyperplasias, 10 chronic prostatitides, and 35 prostate cancer) who underwent prostate needle biopsy. After obtaining serum under suitable conditions, RNA isolation, cDNA synthesis, and qRT-PCR analysis were performed using Qiagen brand kits on Rotor-Gene® Q (Qiagen, Germany) device. -∆Ct values were calculated using RNU6 as a reference gene for normalization. -∆Ct values were used in all statistical calculations.Results: It was observed that miR-21 serum levels were upregulated in chronic prostatitis and cancer groups compared to benign prostatic hyperplasia and the difference between the groups was statistically significant (p = 0.021 and p = 0.001, respectively). The specificity and sensitivity of miR-21 and miR-21/miR-34a combination was calculated as 56% and 86%; 84% and 71% in discriminating benign prostatic hyperplasia and prostate cancer groups, respectively. Conclusion: In this study, it has been shown that miR-21 and miR-21/miR-34a combination has diagnostic performance that can be a biomarker candidate in diagnosing prostate cancer. In addition, the presence of a gradual increase in chronic prostatitis and prostate cancer at miR-21 levels compared to benign prostatic hyperplasia suggests that inflammation and cancer transformation processes taking place at the molecular level are also reflected in the circulating microRNA profile.

Keywords:

Prostate cancer, Prostatitis, Benign prostatic hyperplasia, MicroRNA, Diagnostic efficiency,

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1. Bray F, Ferlay J, Soerjomataram I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians. 2018; 68(6): 394-424.
2. Hamdy FC, Donovan JL, Lane J, et al. 10-year outcomes after monitoring, surgery, or radiotherapy for localized prostate cancer. The New England Journal of Medicine. 2016; 375: 1415-1424.
3. Helgstrand JT, Røder MA, Klemann N, et al. Trends in incidence and 5‐year mortality in men with newly diagnosed, metastatic prostate cancer—A population‐based analysis of 2 national cohorts. Cancer. 2018; 124(14): 2931-2938.
4. Lowenstein LM, Basourakos SP, Williams MD, et al. Active surveillance for prostate and thyroid cancers: evolution in clinical paradigms and lessons learned. Nature Reviews Clinical Oncology. 2019; 16(3): 168-184.
5. Aghdam AM, Amiri A, Salarinia R, Masoudifar A, et al. MicroRNAs as diagnostic, prognostic, and therapeutic biomarkers in prostate cancer. Critical Reviews in Eukaryotic Gene Expression. 2019; 29(2): 127-139.
6. Jamaspishvili T, Berman DM, Ross AE, et al. Clinical implications of PTEN loss in prostate cancer. Nature Reviews Urology. 2018; 15(4): 222-234.
7. Yang Y, Guo JX, Shao ZQ. miR-21 targets and inhibits tumor suppressor gene PTEN to promote prostate cancer cell proliferation and invasion: An experimental study. Asian Pacific journal of tropical medicine. 2017; 10(1): 87-91.
8. Yan X, Tang B, Chen B, Shan Y, Yang H. Replication Study: The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44. eLife. 2019; 8: e43511.
9. Agaoglu FY, Kovancilar M, Dizdar Y, et al. Investigation of miR-21, miR-141, and miR-221 in blood circulation of patients with prostate cancer. Tumor Biology. 2011; 32(3): 583-588.
10. Watahiki A, Macfarlane RJ, Gleave ME, et al.. Plasma miRNAs as biomarkers to identify patients with castration-resistant metastatic prostate cancer. International journal of molecular sciences. 2013; 14(4): 7757-7770.
11. Thompson IM, Ankerst DP, Chi C, et al. Operating characteristics of prostate-specific antigen in men with an initial PSA level of 3.0 ng/ml or lower. Jama. 2005; 294(1): 66-70.
12. Kotb S, Mosharafa A, Essawi M, et al. Circulating miRNAs 21 and 221 as biomarkers for early diagnosis of prostate cancer. Tumor Biology. 2014; 35(12): 12613-12617.
13. Endzeliņš E, Berger A, Melne V, et al. Detection of circulating miRNAs: comparative analysis of extracellular vesicle-incorporated miRNAs and cell-free miRNAs in whole plasma of prostate cancer patients. BMC cancer. 2017; 17(1): 730.
14. Chen Y, Chen S, Zhang J, et al. Expression profile of microRNAs in expressed prostatic secretion of healthy men and patients with IIIA chronic prostatitis/chronic pelvic pain syndrome. Oncotarget. 2018; 9(15): 12186-12200.
15. Ribas J, Ni X, Haffner M, et al. miR-21: an androgen receptor–regulated microRNA that promotes hormone-dependent and hormone-independent prostate cancer growth. Cancer research. 2009; 69(18): 7165-7169.
16. Fabbri M, Paone A, Calore F, et al. MicroRNAs bind to Toll-like receptors to induce prometastatic inflammatory response. Proceedings of the National Academy of Sciences. 2012; 109(31): E2110-E2116.