PIK3CA mutations are frequently observed in BRCAX but not BRCA2 -associated male breast cancer

Breast Cancer Research

Stephen B Fox¹  Alexander Dobrovic¹  kConFab Investigators³  Nicholas Jene²  David Byrne²  Hongdo Do²  Siddhartha Deb¹ 

[1] Department of Pathology and the Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, VIC 3052, Australia;Department of Pathology, Peter MacCallum Cancer Centre, East Melbourne, VIC 3002, Australia;Kathleen Cuningham Foundation Consortium for research into Familial Breast Cancer, Peter MacCallum Cancer Centre, East Melbourne, VIC 3002, Australia

关键词: BRCAX;    BRCA2;    male breast cancer;    familial;    mTOR;    E547K;    PIK3CA;   

Others  :  793782
DOI  :  10.1186/bcr3463

 received in 2012-11-30, accepted in 2013-08-23,  发布年份 2013

【 摘 要 】

Introduction

Although a substantial proportion of male breast cancers (MBCs) are hereditary, the molecular pathways that are activated are unknown. We therefore examined the frequency and clinicopathological associations of the PIK3CA/mammalian target of rapamycin (mTOR) and mitogen-activated protein kinase (MAPK) pathways and their regulatory genes in familial MBC.

Methods

High resolution melting analysis and confirmatory sequencing was used to determine the presence of somatic mutations in PIK3CA (exon 9 and 20), AKT1 (exon 4), KRAS (exon 2) and BRAF (exon 15) genes in 57 familial MBCs. Further analysis of the PIK3CA/mTOR pathway was performed using immunohistochemistry for the pAKT1, pS6 and p4EBP1 biomarkers.

Results

PIK3CA somatic mutations were identified in 10.5% (6 of 57) of cases; there were no AKT1, KRAS or BRAF somatic mutations. PIK3CA mutations were significantly more frequent in cancers from BRCAX patients (17.2%, 5/29) than BRCA2 (0%, 0/25) carriers (P = 0.030). Two BRCAX patients had an E547K mutation which has only been reported in one female breast cancer previously. PIK3CA mutation was significantly correlated with positive pS6 (83.3% vs. 32.0%, P = 0.024) and negative p4EBP1 (100% vs. 38.0%, P = 0.006) expression, but not pAKT expression. Expression of nuclear p4EBP1 correlated with BRCA2 mutation carrier status (68.0% vs. 38.7%, P = 0.035).

Conclusions

Somatic PIK3CA mutation is present in familial male breast cancer but absent in BRCA2 carriers. The presence of two of the extremely rare E547K PIK3CA mutations in our cohort may have specific relevance in MBCs. Further study of PIK3CA in MBCs, and in particular BRCAX patients, may contribute to further establishing the relevance of specific PIK3CA mutations in MBC aetiology and in the identification of particular patient groups most likely to benefit from therapeutic targeting with the novel PIK3CA inhibitors that are currently in development.

【 授权许可】

   
2013 Deb et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Deb S, Jene N, Kconfab Investigators, Fox SB: Genotypic and phenotypic analysis of male breast cancer shows under representation of the HER2 and basal subtypes in BRCA-associated carcinomas. BMC Cancer 2012, 12:510. BioMed Central Full Text
• [2]Nilsson C, Holmqvist M, Bergkvist L, Hedenfalk I, Lambe M, Fjällskog ML: Similarities and differences in the characteristics and primary treatment of breast cancer in men and women - a population based study (Sweden). Acta Oncol 2011, 50:1083-1088.
• [3]Campbell IG, Russell SE, Choong DY, Montgomery KG, Ciavarella ML, Hooi CS, Cristiano BE, Pearson RB, Phillips WA: Mutation of the PIK3CA gene in ovarian and breast cancer. Cancer Res 2004, 64:7678-7681.
• [4]Levine DA, Bogomolniy F, Yee CJ, Lash A, Barakat RR, Borgen PI, Boyd J: Frequent mutation of the PIK3CA gene in ovarian and breast cancers. Clin Cancer Res 2005, 11:2875-2878.
• [5]Ikenoue T, Kanai F, Hikiba Y, Obata T, Tanaka Y, Imamura J, Ohta M, Jazag A, Guleng B, Tateishi K, Asaoka Y, Matsumura M, Kawabe T, Omata M: Functional analysis of PIK3CA gene mutations in human colorectal cancer. Cancer Res 2005, 65:4562-4567.
• [6]Isakoff SJ, Engelman JA, Irie HY, Luo J, Brachmann SM, Pearline RV, Cantley LC, Brugge JS: Breast cancer-associated PIK3CA mutations are oncogenic in mammary epithelial cells. Cancer Res 2005, 65:10992-11000.
• [7]Chen B, Tardell C, Higgins B, Packman K, Boylan JF, Niu H: BRAFV600E negatively regulates the AKT pathway in melanoma cell lines. PloS One 2012, 7:e42598.
• [8]Chu PY, Li TK, Ding ST, Lai IR, Shen TL: EGF-induced Grb7 recruits and promotes Ras activity essential for the tumorigenicity of Sk-Br3 breast cancer cells. J Biol Chem 2010, 285:29279-29285.
• [9]Downward J: Targeting RAS signalling pathways in cancer therapy. Nat Rev Cancer 2003, 3:11-22.
• [10]Junttila MR, Li SP, Westermarck J: Phosphatase-mediated crosstalk between MAPK signaling pathways in the regulation of cell survival. FASEB J 2008, 22:954-965.
• [11]Kiyatkin A, Aksamitiene E, Markevich NI, Borisov NM, Hoek JB, Kholodenko BN: Scaffolding protein Grb2-associated binder 1 sustains epidermal growth factor-induced mitogenic and survival signaling by multiple positive feedback loops. J Biol Chem 2006, 281:19925-19938.
• [12]Kubicek M, Pacher M, Abraham D, Podar K, Eulitz M, Baccarini M: Dephosphorylation of Ser-259 regulates Raf-1 membrane association. J Biol Chem 2002, 277:7913-7919.
• [13]Rodriguez-Viciana P, Warne PH, Dhand R, Vanhaesebroeck B, Gout I, Fry MJ, Waterfield MD, Downward J: Phosphatidylinositol-3-OH kinase as a direct target of Ras. Nature 1994, 370:527-532.
• [14]Buttitta F, Felicioni L, Barassi F, Martella C, Paolizzi D, Fresu G, Salvatore S, Cuccurullo F, Mezzetti A, Campani D, Marchetti A: PIK3CA mutation and histological type in breast carcinoma: high frequency of mutations in lobular carcinoma. J Pathol 2006, 208:350-355.
• [15]Michelucci A, Di Cristofano C, Lami A, Collecchi P, Caligo A, Decarli N, Leopizzi M, Aretini P, Bertacca G, Porta RP, Ricci S, Della Rocca C, Stanta G, Bevilacqua G, Cavazzana A: PIK3CA in breast carcinoma: a mutational analysis of sporadic and hereditary cases. Diagn Mol Pathol 2009, 18:200-205.
• [16]Saal LH, Holm K, Maurer M, Memeo L, Su T, Wang X, Yu JS, Malmstrom PO, Mansukhani M, Enoksson J, Hibshoosh H, Borg A, Parsons R: PIK3CA mutations correlate with hormone receptors, node metastasis, and ERBB2, and are mutually exclusive with PTEN loss in human breast carcinoma. Cancer Res 2005, 65:2554-2559.
• [17]Stemke-Hale K, Gonzalez-Angulo AM, Lluch A, Neve RM, Kuo WL, Davies M, Carey M, Hu Z, Guan Y, Sahin A, Symmans WF, Pusztai L, Nolden LK, Horlings H, Berns K, Hung MC, van de Vijver MJ, Valero V, Gray JW, Bernards R, Mills GB, Hennessy BT: An integrative genomic and proteomic analysis of PIK3CA, PTEN, and AKT mutations in breast cancer. Cancer Res 2008, 68:6084-6091.
• [18]Ibrahim YH, García-García C, Serra V, He L, Torres-Lockhart K, Prat A, Anton P, Cozar P, Guzmán M, Grueso J, Rodríguez O, Calvo MT, Aura C, Díez O, Rubio IT, Pérez J, Rodón J, Cortés J, Ellisen LW, Scaltriti M, Baselga J: PI3K inhibition impairs BRCA1/2 expression and sensitizes BRCA-proficient triple-negative breast cancer to PARP inhibition. Cancer Discov 2012, 2:1036-1047.
• [19]Benvenuti S, Frattini M, Arena S, Zanon C, Cappelletti V, Coradini D, Daidone MG, Pilotti S, Pierotti MA, Bardelli A: PIK3CA cancer mutations display gender and tissue specificity patterns. Hum Mutat 2008, 29:284-288.
• [20]Dawson PJ, Schroer KR, Wolman SR: ras and p53 genes in male breast cancer. Mod Pathol 1996, 9:367-370.
• [21]Hiort O, Naber SP, Lehners A, Muletta-Feurer S, Sinnecker GH, Zöllner A, Komminoth P: The role of androgen receptor gene mutations in male breast carcinoma. J Clin Endocrinol Metab 1996, 81:3404-3407.
• [22]Kwiatkowska E, Teresiak M, Breborowicz D, Mackiewicz A: Somatic mutations in the BRCA2 gene and high frequency of allelic loss of BRCA2 in sporadic male breast cancer. Int J Cancer 2002, 98:943-945.
• [23]Anelli A, Anelli TF, Youngson B, Rosen PP, Borgen PI: Mutations of the p53 gene in male breast cancer. Cancer 1995, 75:2233-2238.
• [24]Johansson I, Nilsson C, Berglund P, Lauss M, Ringner M, Olsson H, Luts L, Sim E, Thorstensson S, Fjallskog ML, Hedenfalk I: Gene expression profiling of primary male breast cancers reveals two unique subgroups and identifies N-acetyltransferase-1 (NAT1) as a novel prognostic biomarker. Breast Cancer Res 2012, 14:R31. BioMed Central Full Text
• [25]Johansson I, Nilsson C, Berglund P, Strand C, Jönsson G, Staaf J, Ringnér M, Nevanlinna H, Barkardottir RB, Borg A, Olsson H, Luts L, Fjällskog ML, Hedenfalk I: High-resolution genomic profiling of male breast cancer reveals differences hidden behind the similarities with female breast cancer. Breast Cancer Res Treat 2011, 129:747-760.
• [26]Callari M, Cappelletti V, De Cecco L, Musella V, Miodini P, Veneroni S, Gariboldi M, Pierotti MA, Daidone MG: Gene expression analysis reveals a different transcriptomic landscape in female and male breast cancer. Breast Cancer Res Treat 2011, 127:601-610.
• [27]Blanco A, de la Hoya M, Balmana J, Ramon y Cajal T, Teule A, Miramar MD, Esteban E, Infante M, Benitez J, Torres A, Tejada MI, Brunet J, Grana B, Balbin M, Perez-Segura P, Osorio A, Velasco EA, Chirivella I, Calvo MT, Feliubadalo L, Lasa A, Diez O, Carracedo A, Caldes T, Vega A: Detection of a large rearrangement in PALB2 in Spanish breast cancer families with male breast cancer. Breast Cancer Res Treat 2012, 132:307-315.
• [28]Falchetti M, Lupi R, Rizzolo P, Ceccarelli K, Zanna I, Calo V, Tommasi S, Masala G, Paradiso A, Gulino A, Giannini G, Russo A, Palli D, Ottini L: BRCA1/BRCA2 rearrangements and CHEK2 common mutations are infrequent in Italian male breast cancer cases. Breast Cancer Res Treat 2008, 110:161-167.
• [29]Orr N, Lemnrau A, Cooke R, Fletcher O, Tomczyk K, Jones M, Johnson N, Lord CJ, Mitsopoulos C, Zvelebil M, McDade SS, Buck G, Blancher C, Trainer AH, James PA, Bojesen SE, Bokmand S, Nevanlinna H, Mattson J, Friedman E, Laitman Y, Palli D, Masala G, Zanna I, Ottini L, Giannini G, Hollestelle A, Ouweland AM, Novakovic S, Krajc M, et al.: Genome-wide association study identifies a common variant in RAD51B associated with male breast cancer risk. Nat Genet 2012, 44:1182-1184.
• [30]Sauty de Chalon A, Teo Z, Park DJ, Odefrey FA, Hopper JL, Southey MC: Are PALB2 mutations associated with increased risk of male breast cancer? Breast Cancer Res Treat 2010, 121:253-255.
• [31]Silvestri V, Rizzolo P, Falchetti M, Zanna I, Masala G, Bianchi S, Palli D, Ottini L: Mutation analysis of BRIP1 in male breast cancer cases: a population-based study in Central Italy. Breast Cancer Res Treat 2011, 126:539-543.
• [32]Wasielewski M, den Bakker MA, van den Ouweland A, Meijer-van Gelder ME, Portengen H, Klijn JG, Meijers-Heijboer H, Foekens JA, Schutte M: CHEK2 1100delC and male breast cancer in the Netherlands. Breast Cancer Res Treat 2009, 116:397-400.
• [33]Davies H, Bignell GR, Cox C, Stephens P, Edkins S, Clegg S, Teague J, Woffendin H, Garnett MJ, Bottomley W, Davis N, Dicks E, Ewing R, Floyd Y, Gray K, Hall S, Hawes R, Hughes J, Kosmidou V, Menzies A, Mould C, Parker A, Stevens C, Watt S, Hooper S, Wilson R, Jayatilake H, Gusterson BA, Cooper C, Shipley J, et al.: Mutations of the BRAF gene in human cancer. Nature 2002, 417:949-954.
• [34]Karnoub AE, Weinberg RA: Ras oncogenes: split personalities. Nat Rev Mol Cell Biol 2008, 9:517-531.
• [35]kConFab: Kathleen Cuningham Foundation Consortium for research into Familial Breast Cancer [http://www.kconfab.org] webcite
• [36]Antoniou AC, Hardy R, Walker L, Evans DG, Shenton A, Eeles R, Shanley S, Pichert G, Izatt L, Rose S, Douglas F, Eccles D, Morrison PJ, Scott J, Zimmern RL, Easton DF, Pharoah PD: Predicting the likelihood of carrying a BRCA1 or BRCA2 mutation: validation of BOADICEA, BRCAPRO, IBIS, Myriad and the Manchester scoring system using data from UK genetics clinics. J Med Genet 2008, 45:425-431.
• [37]Mann GJ, Thorne H, Balleine RL, Butow PN, Clarke CL, Edkins E, Evans GM, Fereday S, Haan E, Gattas M, Giles GG, Goldblatt J, Hopper JL, Kirk J, Leary JA, Lindeman G, Niedermayr E, Phillips KA, Picken S, Pupo GM, Saunders C, Scott CL, Spurdle AB, Suthers G, Tucker K, Chenevix-Trench G: Analysis of cancer risk and BRCA1 and BRCA2 mutation prevalence in the kConFab familial breast cancer resource. Breast Cancer Res 2006, 8:R12. BioMed Central Full Text
• [38]McShane LM, Altman DG, Sauerbrei W, Taube SE, Gion M, Clark GM: Reporting recommendations for tumor marker prognostic studies. J Clin Oncol 2005, 23:9067-9072.
• [39]Lakhani SR, Ellis IO, Schnitt SJ, Tan PH, van de Vijver MJE: WHO Classification of Tumours of the Breast. Lyon: World Health Organization, IARC; 2012.
• [40]Nielsen TO, Hsu FD, Jensen K, Cheang M, Karaca G, Hu Z, Hernandez-Boussard T, Livasy C, Cowan D, Dressler L, Akslen LA, Ragaz J, Gown AM, Gilks CB, van de Rijn M, Perou CM: Immunohistochemical and clinical characterization of the basal-like subtype of invasive breast carcinoma. Clin Cancer Res 2004, 10:5367-5374.
• [41]Do H, Dobrovic A: Dramatic reduction of sequence artefacts from DNA isolated from formalin-fixed cancer biopsies by treatment with uracil- DNA glycosylase. Oncotarget 2012, 3:546-558.
• [42]Do H, Krypuy M, Mitchell PL, Fox SB, Dobrovic A: High resolution melting analysis for rapid and sensitive EGFR and KRAS mutation detection in formalin fixed paraffin embedded biopsies. BMC Cancer 2008, 8:142. BioMed Central Full Text
• [43]Krypuy M, Newnham GM, Thomas DM, Conron M, Dobrovic A: High resolution melting analysis for the rapid and sensitive detection of mutations in clinical samples: KRAS codon 12 and 13 mutations in non-small cell lung cancer. BMC Cancer 2006, 6:295. BioMed Central Full Text
• [44]Hollestelle A, Nagel JH, Smid M, Lam S, Elstrodt F, Wasielewski M, Ng SS, French PJ, Peeters JK, Rozendaal MJ, Riaz M, Koopman DG, Ten Hagen TL, de Leeuw BH, Zwarthoff EC, Teunisse A, van der Spek PJ, Klijn JG, Dinjens WN, Ethier SP, Clevers H, Jochemsen AG, den Bakker MA, Foekens JA, Martens JW, Schutte M: Distinct gene mutation profiles among luminal-type and basal-type breast cancer cell lines. Breast Cancer Res Treat 2010, 121:53-64.
• [45]Garcia-Solano J, Conesa-Zamora P, Carbonell P, Trujillo-Santos J, Torres-Moreno DD, Pagan-Gomez I, Rodriguez-Braun E, Perez-Guillermo M: Colorectal serrated adenocarcinoma shows a different profile of oncogene mutations, MSI status and DNA repair protein expression compared to conventional and sporadic MSI-H carcinomas. Int J Cancer 2012, 131:1790-1799.
• [46]Capodanno A, Boldrini L, Ali G, Pelliccioni S, Mussi A, Fontanini G: Phosphatidylinositol-3-kinase alpha catalytic subunit gene somatic mutations in bronchopulmonary neuroendocrine tumours. Oncol Rep 2012, 28:1559-1566.
• [47]Liao X, Morikawa T, Lochhead P, Imamura Y, Kuchiba A, Yamauchi M, Nosho K, Qian ZR, Nishihara R, Meyerhardt JA, Fuchs CS, Ogino S: Prognostic role of PIK3CA mutation in colorectal cancer: cohort study and literature review. Clin Cancer Res 2012, 18:2257-2268.
• [48]Samuels Y, Wang Z, Bardelli A, Silliman N, Ptak J, Szabo S, Yan H, Gazdar A, Powell SM, Riggins GJ, Willson JK, Markowitz S, Kinzler KW, Vogelstein B, Velculescu VE: High frequency of mutations of the PIK3CA gene in human cancers. Science 2004, 304:554.
• [49]Ottini L, Silvestri V, Rizzolo P, Falchetti M, Zanna I, Saieva C, Masala G, Bianchi S, Manoukian S, Barile M, Peterlongo P, Varesco L, Tommasi S, Russo A, Giannini G, Cortesi L, Viel A, Montagna M, Radice P, Palli D: Clinical and pathologic characteristics of BRCA-positive and BRCA-negative male breast cancer patients: results from a collaborative multicenter study in Italy. Breast Cancer Res Treat 2012, 134:411-418.
• [50]Kimbung S, Biskup E, Johansson I, Aaltonen K, Ottosson-Wadlund A, Gruvberger-Saal S, Cunliffe H, Fadeel B, Loman N, Berglund P, Hedenfalk I: Co-targeting of the PI3K pathway improves the response of BRCA1 deficient breast cancer cells to PARP1 inhibition. Cancer Lett 2012, 319:232-241.
• [51]Basham VM, Lipscombe JM, Ward JM, Gayther SA, Ponder BA, Easton DF, Pharoah PD: BRCA1 and BRCA2 mutations in a population-based study of male breast cancer. Breast Cancer Res 2002, 4:R2. BioMed Central Full Text
• [52]Evans DG, Bulman M, Young K, Howard E, Bayliss S, Wallace A, Lalloo F: BRCA1/2 mutation analysis in male breast cancer families from North West England. Fam Cancer 2008, 7:113-117.
• [53]Korde LA, Zujewski JA, Kamin L, Giordano S, Domchek S, Anderson WF, Bartlett JM, Gelmon K, Nahleh Z, Bergh J, Cutuli B, Pruneri G, McCaskill-Stevens W, Gralow J, Hortobagyi G, Cardoso F: Multidisciplinary meeting on male breast cancer: summary and research recommendations. J Clin Oncol 2010, 28:2114-2122.
• [54]Ottini L, Rizzolo P, Zanna I, Falchetti M, Masala G, Ceccarelli K, Vezzosi V, Gulino A, Giannini G, Bianchi S, Sera F, Palli D: BRCA1/BRCA2 mutation status and clinical-pathologic features of 108 male breast cancer cases from Tuscany: a population-based study in central Italy. Breast Cancer Res Treat 2009, 116:577-586.
• [55]Comprehensive molecular portraits of human breast tumours Nature 2012, 490:61-70.
• [56]Bhagwat SV, Gokhale PC, Crew AP, Cooke A, Yao Y, Mantis C, Kahler J, Workman J, Bittner M, Dudkin L, Epstein DM, Gibson NW, Wild R, Arnold LD, Houghton PJ, Pachter JA: Preclinical characterization of OSI-027, a potent and selective inhibitor of mTORC1 and mTORC2: distinct from rapamycin. Mol Cancer Ther 2011, 10:1394-1406.