Deregulation of miR-100, miR-99a and miR-199b in tissues and plasma coexists with increased expression of mTOR kinase in endometrioid endometrial carcinoma

BMC Cancer

Anna Torres⁵  Kamil Torres⁷  Anna Pesci¹  Marcello Ceccaroni²  Tomasz Paszkowski⁵  Paola Cassandrini⁴  Giuseppe Zamboni³  Ryszard Maciejewski⁶ 

[1] Department of Pathology, Ospedale Sacro Cuore Don Calabria, Via don A. Sempreboni 5, Negrar, Verona, Italy

[2] Department of Obstetrics & Gynecology, European Gynecology Endoscopy School, Ospedale Sacro Cuore Don Calabria, Via don A. Sempreboni 5, Negrar, Verona, Italy

[3] Department of Pathology, University of Verona, Piazzale L.A. Scuro 10, 37134, Verona, Italy

[4] Department of Oncology, Ospedale Sacro Cuore Don Calabria, Via don A. Sempreboni 5, Negrar, Verona, Italy

[5] III Chair and Department of Gynecology, Medical University of Lublin, Jaczewskiego 8, Lublin, Poland

[6] II Chair and Department of Surgery, Medical University of Lublin, Staszica 16, 20-081, Lublin, Poland

[7] General and Oncologic Surgery Department, Lublin County Specialist Hospital, Al. Kraśnicka 100, Lublin, Poland

关键词: Plasma;    Endometrial cancer;    miRNA;    microRNA;    mTOR;   

Others  :  1080239
DOI  :  10.1186/1471-2407-12-369

 received in 2012-05-25, accepted in 2012-08-16,  发布年份 2012

【 摘 要 】

Background

Alterations of mTOR gene expression have been implicated in the pathogenesis of endometrioid endometrial cancer however only few studies explored the cause of increased mTOR activation in this malignancy. miRNAs are small, noncoding RNAs, which were proven to regulated gene expression at the posttranscriptional level. The study aimed to explore deregulation of miRNAs targeting mTOR kinase (miR-99a, miR-100 and miR-199b) as a possible cause of its altered expression in EEC tissues. In addition expression of the three miRNAs was investigated in plasma of EEC patients and was assessed in terms of diagnostic and prognostic utility.

Methods

We investigated expression of mTOR kinase transcripts in 46 fresh tissue samples. Expression of miR-99a, miR-100 and miR-199b was investigated in the same group of fresh samples, and in additional 58 FFPE sections as well as in 48 plasma samples using qPCR. Relative quantification was performed using experimentally validated endogenous controls.

Results

mTOR kinase expression was increased in EEC tissues and was accompanied by decreased expression of all three miRNAs. Down-regulation of the investigated miRNAs was discovered in plasma of EEC patients and miRNA signatures classified EEC tissues (miR-99a/miR-100/miR-199b) and plasma (miR-99a/miR-199b) samples with higher accuracy in comparison to single miRNAs. We also revealed that miR-100 was an independent prognostic marker of overall survival.

Conclusions

We conclude that increased expression of mTOR kinase coexists with down-regulation of its targeting miRNAs, which could suggest a new mechanism of mTOR pathway alterations in EEC. In addition, our findings implicate that miRNA signatures can be considered promising biomarkers for early detection and prognosis of endometrioid endometrial carcinoma.

【 授权许可】

   
2012 Torres et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

【 参考文献 】

• [1]Ferlay J, Parkin DM, Steliarova-Foucher E: Estimates of cancer incidence and mortality in Europe in 2008. Eur J Cancer 2010, 6:765-781.
• [2]Siegel R, Ward E, Brawley O, Jemal A: Cancer statistics, 2011: the impact of eliminating socioeconomic and racial disparities on premature cancer deaths. CA Cancer J Clin 2011, 61:212-236.
• [3]Evans T, Sany O, Pearmain P, Ganesan R, Blann A, Sundar S: Differential trends in the rising incidence of endometrial cancer by type: data from a UK population-based registry from 1994 to 2006. Br J Cancer 2011, 104:1505-1510.
• [4][Polish Cancer Registry] Krajowy Rejestr Nowotworówhttp://epid.coi.waw.pl/krn/ webcite
• [5]Bansal N, Yendluri V, Wenham RM: The molecular biology of endometrial cancers and the implications for pathogenesis, classification, and targeted therapies. Cancer Control 2009, 16:8-13.
• [6]Moreno-Bueno G, Hardisson D, Sarrió D, Sánchez C, Cassia R, Prat J, Herman JG, Esteller M, Matías-Guiu X, Palacios J: Abnormalities of E- and P-cadherin and catenin (beta-, gamma-catenin, and p120ctn) expression in endometrial cancer and endometrial atypical hyperplasia. J Pathol 2003, 199:471-478.
• [7]Velasco A, Bussaglia E, Pallares J, Dolcet X, Llobet D, Encinas M, Llecha N, Palacios J, Prat J, Matias-Guiu X: PIK3CA gene mutations in endometrial carcinoma: correlation with PTEN and K-RAS alterations. Hum Pathol 2006, 37:1465-1472.
• [8]Salvesen HB, Stefansson I, Kretzschmar EI, Gruber P, MacDonald ND, Ryan A, Jacobs IJ, Akslen LA, Das S: Significance of PTEN alterations in endometrial carcinoma: a population-based study of mutations, promoter methylation and PTEN protein expression. Int J Oncol 2004, 25:1615-1623.
• [9]Kong D, Suzuki A, Zou TT, Sakurada A, Kemp LW, Wakatsuki S, Yokoyama T, Yamakawa H, Furukawa T, Sato M, Ohuchi N, Sato S, Yin J, Wang S, Abraham JM, Souza RF, Smolinski KN, Meltzer SJ, Horii A: PTEN1 is frequently mutated in primary endometrial carcinomas. Nat Genet 1997, 17:143-144.
• [10]Mutter GL, Lin MC, Fitzgerald JT, Kum JB, Baak JP, Lees JA, Weng LP, Eng C: Altered PTEN expression as a diagnostic marker for the earliest endometrial precancers. J Natl Cancer Inst 2000, 92:924-930.
• [11]Wullschleger S, Loewith R, Hall MN: TOR signaling in growth and metabolism. Cell 2006, 124:471-484.
• [12]Hay N, Sonenberg N: Upstream and downstream of mTOR. Genes Dev 2004, 18:1926-1945.
• [13]Kim DH, Sarbassov DD, Ali SM, King JE, Latek RR, Erdjument-Bromage H, Tempst P, Sabatini DM: mTOR interacts with raptor to form a nutrient-sensitive complex that signals to the cell growth machinery. Cell 2002, 110:163-175.
• [14]Tokunaga C, Yoshino K, Yonezawa K: mTOR integrates amino acid- and energy-sensing pathways. Biochem Biophys Res Commun 2004, 313:443-446.
• [15]Guertin DA, Sabatini DM: Defining the role of mTOR in cancer. Cancer Cell 2007, 12:9-22.
• [16]Darb-Esfahani S, Faggad A, Noske A, Weichert W, Buckendahl AC, Müller B, Budczies J, Röske A, Dietel M, Denkert C: Phospho-mTOR and phospho-4EBP1 in endometrial adenocarcinoma: association with stage and grade in vivo and link with response to rapamycin treatment in vitro. J Cancer Res Clin Oncol 2009, 135:933-941.
• [17]Huang J, Manning BD: A complex interplay between Akt, TSC2 and the two mTOR complexes. Biochem Soc Trans 2009, 37:217-222.
• [18]Gwinn DM, Shackelford DB, Egan DF, Mihaylova MM, Mery A, Vasquez DS, Turk BE, Shaw RJ: AMPK phosphorylation of raptor mediates a metabolic checkpoint. Mol Cell 2008, 30:214-226.
• [19]Budanov AV, Karin M: p53 target genes sestrin1 and sestrin2 connect genotoxic stress and mTOR signaling. Cell 2008, 134:451-460.
• [20]Shen Q, Stanton ML, Feng W, Rodriguez ME, Ramondetta L, Chen L, Brown RE, Duan X: Morphoproteomic analysis reveals an overexpressed and constitutively activated phospholipase D1-mTORC2 pathway in endometrial carcinoma. Int J Clin Exp Pathol 2010, 4:13-21.
• [21]Sayed D, Abdellatif M: MicroRNAs in development and disease. Physiol Rev 2011, 91:827-887.
• [22]Farazi TA, Spitzer JI, Morozov P, Tuschl T: miRNAs in human cancer. J Pathol 2011, 223:102-115.
• [23]Guo H, Ingolia NT, Weissman JS, Bartel DP: Mammalian microRNAs predominantly act to decrease target mRNA levels. Nature 2010, 466:835-840.
• [24]Lewin SN: Revised FIGO staging system for endometrial cancer. Clin Obstet Gynecol 2011, 54:215-218.
• [25]Pfaffl MW: A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 2001, 29:e45.
• [26]Peltier HJ, Latham GJ: Normalization of microRNA expression levels in quantitative RT-PCR assays: identification of suitable reference RNA targets in normal and cancerous human solid tissues. RNA 2008, 14:844-852.
• [27]Wong L, Lee K, Russell I, Chen C: Endogenous Controls for Real-Time Quantitation of miRNA Using TaqMan® MicroRNA Assays. Applied Biosystems Application Note 2007. Publication 127AP11-01 http://www.appliedbiosystems.com webcite
• [28]Liang Y, Ridzon D, Wong L, Chen C: Characterization of microRNA expression profiles in normal human tissues. BMC Genomics 2007, 8:166. BioMed Central Full Text
• [29]Bargaje R, Hariharan M, Scaria V, Pillai B: Consensus miRNA expression profiles derived from interplatform normalization of microarray data. RNA 2010, 16:16-25.
• [30]Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, Pogosova-Agadjanyan EL, Peterson A, Noteboom J, O'Briant KC, Allen A, Lin DW, Urban N, Drescher CW, Knudsen BS, Stirewalt DL, Gentleman R, Vessella RL, Nelson PS, Martin DB: Tewari: Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci U S A 2008, 105:10513-8.
• [31]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.
• [32]Chen X, Bonnefoi H, Diebold-Berger S, Lyautey J, Lederrey C, Faltin-Traub E, Stroun M, Anker P: Detecting tumor-related alterations in plasma or serum DNA of patients diagnosed with breast cancer. Clin Cancer Res 1999, 5:2297-303.
• [33]Resnick KE, Alder H, Hagan JP, Richardson DL, Croce CM, Cohn DE: The detection of differentially expressed microRNAs from the serum of ovarian cancer patients using a novel real-time PCR platform. Gynecol Oncol 2009, 112:55-9.
• [34]Lawrie CH, Gal S, Dunlop HM, Pushkaran B, Liggins AP, Pulford K, Banham AH, Pezzella F, Boultwood J, Wainscoat JS, Hatton CS, Harris AL: Detection of elevated levels of tumour-associated microRNAs in serum of patients with diffuse large B-cell lymphoma. Br J Haematol 2008, 141:672-5.
• [35]Profiling of microRNA in Blood, Serum/Plasma. http://www.exiqon.com/ls/Documents/Scientific/microRNA-serum-plasma-guidelines.pdf webcite
• [36]Andersen CL, Jensen JL, Ørntoft TF: Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 2004, 64:5245-50.
• [37]Hellemans J, Mortier G, De Paepe A, Speleman F, Vandesompele J: qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol 2007, 8:R19. BioMed Central Full Text
• [38]Hosmer DW, Lemeshow S: Applied logistic regression. New York, NY: Wiley-Interscience; 1989.
• [39]Lu KH, Wu W, Dave B, Slomovitz BM, Burke TW, Munsell MF, Broaddus RR, Walker CL: Loss of tuberous sclerosis complex-2 function and activation of mammalian target of rapamycin signaling in endometrial carcinoma. Clin Cancer Res 2008, 14:2543-50.
• [40]Li X, Xiao L, Yang Y, Shen H, Zeng H, Wang Z: The expression of mammalian target of rapamycin in Ishikawa and HEC-1A cells. J Huazhong Univ Sci Technolog Med Sci 2008, 28:340-2.
• [41]Yoshida Y, Kurokawa T, Horiuchi Y, Sawamura Y, Shinagawa A, Kotsuji F: Localisation of phosphorylated mTOR expression is critical to tumour progression and outcomes in patients with endometrial cancer. Eur J Cancer 2010, 46:3445-52.
• [42]Wahl H, Daudi S, Kshirsagar M, Griffith K, Tan L, Rhode J, Liu JR: Expression of metabolically targeted biomarkers in endometrial carcinoma. Gynecol Oncol 2010, 116:21-7.
• [43]Mori N, Kyo S, Sakaguchi J, Mizumoto Y, Ohno S, Maida Y, Hashimoto M, Takakura M, Inoue M: Concomitant activation of AKT with extracellular-regulated kinase 1/2 occurs independently of PTEN or PIK3CA mutations in endometrial cancer and may be associated with favorable prognosiss. Cancer Sci 2007, 98:1881-8.
• [44]Gadducci A, Cosio S, Genazzani AR: Old and new perspectives in the pharmacological treatment of advanced or recurrent endometrial cancer: Hormonal therapy, chemotherapy and molecularly targeted therapies. Crit Rev Oncol Hematol 2006, 58:242-56.
• [45]Slomovitz BM, Lu KH, Johnston T, Coleman RL, Munsell M, Broaddus RR, Walker C, Ramondetta LM, Burke TW, Gershenson DM, Wolf J: A phase 2 study of the oral mammalian target of rapamycin inhibitor, everolimus, in patients with recurrent endometrial carcinoma. Cancer 2010, 116:5415-9.
• [46]Oza AM, Elit L, Tsao MS, Kamel-Reid S, Biagi J, Provencher DM, Gotlieb WH, Hoskins PJ, Ghatage P, Tonkin KS, Mackay HJ, Mazurka J, Sederias J, Ivy P, Dancey JE, Eisenhauer EA: Phase II study of temsirolimus in women with recurrent or metastatic endometrial cancer: a trial of the NCIC Clinical Trials Group. J Clin Oncol 2011, 29:3278-85.
• [47]Choi CH, Lee JS, Kim SR, Kim TJ, Lee JW, Kim BG, Bae DS: Clinical significance of pmTOR expression in endometrioid endometrial carcinoma. Eur J Obstet Gynecol Reprod Biol 2010, 153:207-10.
• [48]No JH, Jeon YT, Park IA, Kang D, Kim JW, Park NH, Kang SB, Song YS: Expression of mTOR protein and its clinical significance in endometrial cancer. Med Sci Monit 2009, 15:BR301-305.
• [49]Nagaraja AK, Creighton CJ, Yu Z, Zhu H, Gunaratne PH, Reid JG, Olokpa E, Itamochi H, Ueno NT, Hawkins SM, Anderson ML, Matzuk MM: A link between mir-100 and FRAP1/mTOR in clear cell ovarian cancer. Mol Endocrinol 2010, 24:447-63.
• [50]Li D, Liu X, Lin L, Hou J, Li N, Wang C, Wang P, Zhang Q, Zhang P, Zhou W, Wang Z, Ding G, Zhuang SM, Zheng L, Tao W, Cao X: MicroRNA-99a Inhibits Hepatocellular Carcinoma Growth and Correlates with Prognosis of Patients with Hepatocellular Carcinoma. J Biol Chem 2011, 286:36677-85.
• [51]Chen Z, Jin Y, Yu D, Wang A, Mahjabeen I, Wang C, Liu X, Zhou X: Down-regulation of the microRNA-99 family members in head and neck squamous cell carcinoma. Oral Oncolin press
• [52]Sun D, Lee YS, Malhotra A, Kim HK, Matecic M, Evans C, Jensen RV, Moskaluk CA, Dutta A: miR-99 family of MicroRNAs suppresses the expression of prostate-specific antigen and prostate cancer cell proliferation. Cancer Res 2011, 71:1313-24.
• [53]Garzia L, Andolfo I, Cusanelli E, Marino N, Petrosino G, De Martino D, Esposito V, Galeone A, Navas L, Esposito S, Gargiulo S, Fattet S, Donofrio V, Cinalli G, Brunetti A, Vecchio LD, Northcott PA, Delattre O, Taylor MD, Iolascon A, Zollo M: MicroRNA-199b-5p impairs cancer stem cells through negative regulation of HES1 in medulloblastoma. PLoS One 2009, 4:e4998.
• [54]Wu W, Lin Z, Zhuang Z, Liang X: Expression profile of mammalian microRNAs in endometrioid adenocarcinoma. Eur J Cancer Prev 2009, 18:50-5.
• [55]Snowdon J, Zhang X, Childs T, Tron VA, Feilotter H: The microRNA-200 family is upregulated in endometrial carcinoma. PLoS One 2011, 6:e22828.
• [56]Chung TK, Lau TS, Cheung TH, Yim SF, Lo KW, Siu NS, Chan LK, Yu MY, Kwong J, Doran G, Barroilhet LM, Ng AS, Wong RR, Wang VW, Mok SC, Smith DI, Berkowitz RS, Wong YF: Dysregulation of microRNA-204 mediates migration and invasion of endometrial cancer by regulating FOXC1. Int J Cancer 2012, 130:1036-45.
• [57]Tanaka M, Oikawa K, Takanashi M, Kudo M, Ohyashiki J, Ohyashiki K, Kuroda M: Down-regulation of miR-92 in human plasma is a novel marker for acute leukemia patients. PLoS One 2009, 4:e5532.
• [58]Ohyashiki K, Umezu T, Yoshizawa S, Ito Y, Ohyashiki M, Kawashima H, Tanaka M, Kuroda M, Ohyashiki JH: Clinical impact of down-regulated plasma miR-92a levels in non-Hodgkin's lymphoma. PLoS One 2011, 6:e16408.
• [59]Shigoka M, Tsuchida A, Matsudo T, Nagakawa Y, Saito H, Suzuki Y, Aoki T, Murakami Y, Toyoda H, Kumada T, Bartenschlager R, Kato N, Ikeda M, Takashina T, Tanaka M, Suzuki R, Oikawa K, Takanashi M, Kuroda M: Deregulation of miR-92a expression is implicated in hepatocellular carcinoma development. Pathol Int 2010, 60:351-7.
• [60]Cohn DE, Fabbri M, Valeri N, Alder H, Ivanov I, Liu CG, Croce CM, Resnick KE: Comprehensive miRNA profiling of surgically staged endometrial cancer. Am J Obstet Gynecol 2010, 202:656.e1-8.