MicroRNAs in renal cell carcinoma: diagnostic implications of serum miR-1233 levels

Lena M Wulfken, Rudolf Moritz, Carsten Ohlmann, Stefan Holdenrieder, Volker Jung, Frank Becker, Edwin Herrmann, Gisela Walgenbach-Brünagel, Alexander von Ruecker, Stefan C Müller, Jörg Ellinger, Lena M Wulfken, Rudolf Moritz, Carsten Ohlmann, Stefan Holdenrieder, Volker Jung, Frank Becker, Edwin Herrmann, Gisela Walgenbach-Brünagel, Alexander von Ruecker, Stefan C Müller, Jörg Ellinger

Abstract

Background: MicroRNA expression is altered in cancer cells, and microRNAs could serve as diagnostic/prognostic biomarker for cancer patients. Our study was designed to analyze circulating serum microRNAs in patients with renal cell carcinoma (RCC).

Methodology/principal findings: We first explored microrna expression profiles in tissue and serum using taqman low density arrays in each six malignant and benign samples: Although 109 microRNAs were circulating at higher levels in cancer patients' serum, we identified only 36 microRNAs with up-regulation in RCC tissue and serum of RCC patients. Seven candidate microRNAs were selected for verification based on the finding of up-regulation in serum and tissue of RCC patients: miR-7-1*, miR-93, miR-106b*, miR-210, miR-320b, miR-1233 and miR-1290 levels in serum of healthy controls (n = 30) and RCC (n = 33) patients were determined using quantitative real-time PCR (TaqMan MicroRNA Assays). miR-1233 was increased in RCC patients, and thus validated in a multicentre cohort of 84 RCC patients and 93 healthy controls using quantitative real-time PCR (sensitivity 77.4%, specificity 37.6%, AUC 0.588). We also studied 13 samples of patients with angiomyolipoma or oncocytoma, whose serum miR-1233 levels were similar to RCC patients. Circulating microRNAs were not correlated with clinical-pathological parameters.

Conclusions/significance: MicroRNA levels are distinctly increased in cancer patients, although only a small subset of circulating microRNAs has a tumor-specific origin. We identify circulating miR-1233 as a potential biomarker for RCC patients. Larger-scaled studies are warranted to fully explore the role of circulating microRNAs in RCC.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Flow Chart: Verification of miR-1233…
Figure 1. Flow Chart: Verification of miR-1233 as diagnostic marker of renal cell carcinoma.
Figure 2. Serum microRNA levels in patients…
Figure 2. Serum microRNA levels in patients with renal cell carcinoma, benign renal tumors and healthy controls.
MicroRNAs were profiled in serum and tissue of patients with clear cell renal cell carcinoma (ccRCC) and healthy controls (each n = 6) using the TaqMan Low Density Array; 36 microRNAs were upregulated in RCC tissue and serum of RCC patients (Phase-1). Seven microRNAs were chosen for verification using real-time PCR in a cohort of 33 ccRCC and 30 healthy individuals (Phase-2). In Phase-3, the finding of increased serum miR-1233 levels in RCC patients was confirmed in a multicentre cohort of 84 RCC, 93 healthy and 13 benign renal tumors (BRT). MicroRNA recovery (as determined using quantification of the synthetic miR-39) was similar in the study centers (UKB, UKM, UKS); the level of miR-1233 were somewhat higher in patients from UKM.

References

    1. Jemal A, Bray F, Center MM, Ferlay J, Ward E, et al. Global cancer statistics. CA Cancer J Clin. 2011;61:69–90.
    1. Ljungberg B, Cowan NC, Hanbury DC, Hora M, Kuczyk MA, et al. EAU guidelines on renal cell carcinoma: the 2010 update. Eur Urol. 2010;58:398–406.
    1. Volinia S, Calin GA, Liu CG, Ambs S, Cimmino A, et al. A microRNA expression signature of human solid tumors defines cancer gene targets. Proc Natl Acad Sci U S A. 2006;103:2257–2261.
    1. Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, et al. Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci U S A. 2008;105:10513–10518.
    1. Chim SS, Shing TK, Hung EC, Leung TY, Lau TK, et al. Detection and characterization of placental microRNAs in maternal plasma. Clin Chem. 2008;54:482–490.
    1. Hunter MP, Ismail N, Zhang X, Aguda BD, Lee EJ, et al. Detection of microRNA expression in human peripheral blood microvesicles. e3694-PLoS One. 2008;3
    1. Mahn R, Heukamp LC, Rogenhofer S, von Ruecker A, Muller SC, et al. Circulating microRNAs (miRNA) in Serum of Patients With Prostate Cancer. Urology. 2011;77:1265–1265.
    1. Huang Z, Huang D, Ni S, Peng Z, Sheng W, et al. Plasma microRNAs are promising novel biomarkers for early detection of colorectal cancer. Int J Cancer. 2010;127:118–126.
    1. Heneghan HM, Miller N, Lowery AJ, Sweeney KJ, Newell J, et al. Circulating microRNAs as novel minimally invasive biomarkers for breast cancer. Ann Surg. 2010;251:499–505.
    1. Brase JC, Johannes M, Schlomm T, Falth M, Haese A, et al. Circulating miRNAs are correlated with tumor progression in prostate cancer. Int J Cancer. 2011;128:608–616.
    1. Chen X, Ba Y, Ma L, Cai X, Yin Y, et al. Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res. 2008;18:997–1006.
    1. Lawrie CH, Gal S, Dunlop HM, Pushkaran B, Liggins AP, et al. Detection of elevated levels of tumour-associated microRNAs in serum of patients with diffuse large B-cell lymphoma. Br J Haematol. 2008;141:672–675.
    1. Ng EK, Chong WW, Jin H, Lam EK, Shin VY, et al. Differential expression of microRNAs in plasma of patients with colorectal cancer: a potential marker for colorectal cancer screening. Gut. 2009;58:1375–1381.
    1. Resnick KE, Alder H, Hagan JP, Richardson DL, Croce CM, et al. The detection of differentially expressed microRNAs from the serum of ovarian cancer patients using a novel real-time PCR platform. Gynecol Oncol. 2009;112:55–59.
    1. Tsujiura M, Ichikawa D, Komatsu S, Shiozaki A, Takeshita H, et al. Circulating microRNAs in plasma of patients with gastric cancers. Br J Cancer. 2010;102:1174–1179.
    1. Gottardo F, Liu CG, Ferracin M, Calin GA, Fassan M, et al. Micro-RNA profiling in kidney and bladder cancers. Urol Oncol. 2007;25:387–392.
    1. Nakada C, Matsuura K, Tsukamoto Y, Tanigawa M, Yoshimoto T, et al. Genome-wide microRNA expression profiling in renal cell carcinoma: significant down-regulation of miR-141 and miR-200c. J Pathol. 2008;216:418–427.
    1. Jung M, Mollenkopf HJ, Grimm C, Wagner I, Albrecht M, et al. MicroRNA profiling of clear cell renal cell cancer identifies a robust signature to define renal malignancy. J Cell Mol Med. 2009;13:3918–3928.
    1. Huang Y, Dai Y, Yang J, Chen T, Yin Y, et al. Microarray analysis of microRNA expression in renal clear cell carcinoma. Eur J Surg Oncol. 2009;35:1119–1123.
    1. Petillo D, Kort EJ, Anema J, Furge KA, Yang XJ, et al. MicroRNA profiling of human kidney cancer subtypes. Int J Oncol. 2009;35:109–114.
    1. Yi Z, Fu Y, Zhao S, Zhang X, Ma C. Differential expression of miRNA patterns in renal cell carcinoma and nontumorous tissues. J Cancer Res Clin Oncol. 2010;136:855–862.
    1. Juan D, Alexe G, Antes T, Liu H, Madabhushi A, et al. Identification of a microRNA panel for clear-cell kidney cancer. Urology. 2010;75:835–841.
    1. Chow TF, Youssef YM, Lianidou E, Romaschin AD, Honey RJ, et al. Differential expression profiling of microRNAs and their potential involvement in renal cell carcinoma pathogenesis. Clin Biochem. 2010;43:150–158.
    1. Youssef YM, White NM, Grigull J, Krizova A, Samy C, et al. Accurate Molecular Classification of Kidney Cancer Subtypes Using MicroRNA Signature. Eur Urol. 2011;59:721–730.
    1. White NM, Bao TT, Grigull J, Youssef YM, Girgis, A, et al. miRNA Profiling for Clear Cell Renal Cell Carcinoma: Biomarker Discovery and Identification of Potential Controls and Consequences of miRNA Dysregulation. J Urol. 2011;186:1077–1083.
    1. Ellinger J, El Kassem N, Heukamp LC, Mathews S, Cubukluoz F, et al. Hypermethylation of cell-free serum DNA indicates worse outcome in patients with bladder cancer. J Urol. 2008;179:346–352.
    1. Ellinger J, Bastian PJ, Haan KI, Heukamp LC, Buettner R, et al. Noncancerous PTGS2 DNA fragments of apoptotic origin in sera of prostate cancer patients qualify as diagnostic and prognostic indicators. Int J Cancer. 2008;122:138–143.
    1. Ellinger J, Haan K, Heukamp LC, Kahl P, Buttner R, et al. CpG Island hypermethylation in cell-free serum DNA identifies patients with localized prostate cancer. Prostate. 2008;68:42–49.
    1. Ellinger J, Albers P, Perabo FG, Muller SC, von Ruecker A, et al. CpG Island Hypermethylation of cell-free circulating serum DNA in patients with testicular cancer. J Urol. 2009;182:324–329.
    1. Dweep H, Sticht C, Pandey P, Gretz N. miRWalk - Database: Prediction of possible miRNA binding sites by “walking” the genes of three genomes. J Biomed Inform. 2011.
    1. Moreira JM, Ohlsson G, Gromov P, Simon R, Sauter G, et al. Bladder cancer-associated protein, a potential prognostic biomarker in human bladder cancer. Mol Cell Proteomics. 2010;9:161–177.
    1. Kroh EM, Parkin RK, Mitchell PS, Tewari M. Analysis of circulating microRNA biomarkers in plasma and serum using quantitative reverse transcription-PCR (qRT-PCR). Methods. 2010;50:298–301.

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