Altered MicroRNAExpressionin Cervical Carcinomas Jeong-WonLee, 1 ChelHunChoi, 1 Jung-JooChoi, 1 Young-AePark, 1 Seung-JunKim, 2 Seung YongHwang, 2 Woo YoungKim, 1 Tae-JoongKim, 1 Je-Ho Lee, 1 Byoung-Gie Kim, 1 and Duk-Soo Bae 1 Abstract Purpose: MicroRNAs (miRNA) are small noncoding RNAs that are 18 to 25 nucleotides in length; they regulate the stability or translational efficiency of target mRNAs. Emerging evidence suggeststhatmiRNAsmightbeinvolvedinthepathogenesisofavarietyofhumancancers. Experimental Design: In this study, we profiled miRNA expression in 10 early stage invasive squamous cell carcinomas (ISCC) and 10 normal cervical squamous epithelial specimens using TaqMan real-time quantitative PCRarray methods. Inorder to evaluate the role of miR-199a, one of the most significantly overexpressed in ISCCs, we transfected cervical cancer cells (SiHa and ME-180) with anti^ miR-199a oligonucleotides and assessed the cell viability. Results: We found 70 genes (68 up-regulated, 2 down-regulated) with significantly different expression in the ISCCs compared with normal samples (P < 0.05). When we analyzed the expression of the 10 most significant miRNAs in 31 ISCCs, increased miR-127 expression was significantly associated with lymph node metastasis (P = 0.006). Transfection of anti ^ miR-199a oligonucleotides to cervical cancer cells suppressed cell growth in vitro, which was potentiated with the anticancer agent cisplatin. Conclusions: Our results show that miRNA deregulation may play an important role in the malignant transformation of cervical squamous cells. In addition, they may offer new candidate targets to be exploited for both prognostic and therapeutic strategies in patients with cervical cancer. Cervicalcanceristheresultofamultistepprocessthatinvolves the transformation of the normal cervical epithelium to a preneoplastic cervical intraepithelial neoplasia that is subse- quentlytransformedtoinvasivecervicalcancer(1,2).Although high-risk human papillomaviruses are associated with cervical cancer (1, 2), human papillomavirus infection alone is not sufficienttoinducethemalignanttransformation.Hence,other unidentified genetic alterations are likely involved. The identification of such genetic alterations would be of consid- erable importance for the screening and treatment of cervical cancer (3). MicroRNAs (miRNA) are a recently discovered class of small noncoding RNAs that regulate gene expression (4). Mature 18- to 25-nucleotide-long miRNAs regulate gene expression by catalyzing the cleavage of mRNA (5–10) or repressing mRNA translation (9, 11, 12). The specific roles of miRNAs include the regulation of cell proliferation and metabolism (13, 14), developmental timing (4, 14), cell death (15), hematopoiesis (16), neuron development (17), human tumorigenesis (18), DNA methylation, and chromatin modification (19). There is increasing evidence that the expression of miRNA genesisderegulatedinhumancancer(20–26).AlteredmiRNA expression profiles have been reported in lung cancer (27), breastcancer(28),glioblastoma(29),hepatocellularcarcinoma (30), papillary thyroid carcinoma (31), and more recently, colorectal cancer (26). Moreover, some studies have reported that miRNA expression signatures are associated with clinical outcomes of certain diseases (32–34). These data suggest that miRNAs play an important role in a variety of human cancers. However, the molecular basis of miRNA-mediated gene regulation is not fully understood and their role in tumorigen- esis remains largely unknown. Over the past several years, a number of methods for quantifying miRNAs have been described, including Northern blotting, microarrays (35, 36), a modified invader assay (37), a bead-based flow cytometric assay (22), and real-time quan- titative PCR (38). Real-time quantitative PCR is more quantita- tive and sensitive than other high-throughput assays. Here, we present the results of miRNA deregulation in a set of early stage invasive squamous cell carcinomas (ISCC) and normal cervicalepithelialtissues,usingreal-timequantitativePCRarray methods. Human Cancer Biology Authors’Affiliations: 1 Department of Obstetrics and Gynecology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea and 2 Department of Biochemistry, Hanyang University and GenoCheck Co. Ltd., Gyeonggi-do, Korea Received5/18/07;revised10/17/07;accepted11/7/07. Grant support: Samsung Biomedical Research Institute grant (no. SBRIC-A7- 414-1) and the IN-SUNGFoundation for Medical Research (no. C-A7-804-1). Thecostsofpublicationofthisarticleweredefrayedinpartbythepaymentofpage charges.This article must therefore be hereby marked advertisement in accordance with18U.S.C.Section1734solely toindicatethisfact. Note: Supplementary data for this article are available at Clinical Cancer Research Online(http://clincancerres.aacrjournals.org/). J-W. Lee, C.H. Choi, B-G. Kim, and D-S. Bae contributed equally to this paper. Requests for reprints: Duk-Soo Bae or Byoung-Gie Kim, Department of Obstetrics and Gynecology, Samsung Medical Center, Sungkyunkwan University School of Medicine, 50 Irwon-dong, Gangnam-gu, Seoul 135-710, Korea. Phone: 82-2-3410-3519/3513; Fax: 82-2-3410-0630; E-mail: huna0@naver.com (D-S. Bae)orbksong.kim@samsung.com (B-G. Kim). F 2008AmericanAssociationforCancerResearch. doi:10.1158/1078-0432.CCR-07-1231 www.aacrjournals.org ClinCancerRes2008;14(9)May1,2008 2535 Downloaded from http://aacrjournals.org/clincancerres/article-pdf/14/9/2535/1981794/2535.pdf by guest on 14 June 2022