Alterations of PTEN and SMAD4 methylation in diagnosis of breast cancer: implications of methyl II PCR assay
Tóm tắt
Diagnosis of breast cancer is more complicated due to lack of minimal invasive biomarker with sufficient precision. DNA methylation is a promising marker for cancer diagnosis. In this study, authors evaluated methylation patterns for PTEN and SMAD4 in blood samples using EpiTect Methyl II QPCR assay quantitative PCR technology. Methylation status for PTEN and SMAD4 were statistically significant as breast cancer patients reported hypermethylation compared to benign and control groups (77.1 ± 17.9 vs. 24.9 ± 4.5 and 15.1 ± 1.4 and 70.1 ± 14.4 vs. 28.2 ± 0.61 and 29.5 ± 3.6, respectively). ROC curve analysis revealed that both PTEN (AUC = 0.992) and SMAD4 (AUC = 0.853) had good discriminative power for differentiating BC from all non-cancer individuals (benign and healthy combined) compared to routine tumor markers CEA (AUC = 0.538) and CA15.3 (AUC = 0.686). High PTEN methylation degree was associated with late stages (84.2 ± 17.4), positive lymph node (84.2 ± 18.5), positive ER (81.3 ± 19.7), positive PgR (79.5 ± 19.1), and positive HER2 (80.7 ± 19.0) vs. 67.4 ± 13.8, 70.6 ± 14.8, 72.8 ± 14.9, 72.5 ± 14.7, and 70.2 ± 13.5 in early stages, negative lymph node, negative ER, negative PgR, and negative HER2, respectively. Similar results were obtained regarding SMAD4 methylation. Sensitivity, specificity, positive and negative predictive values, and accuracy for methylated PTEN were 100%, 95%, 99.1%, 100%, and 95%, respectively when differentiated BC from all-non cancer controls. Interestingly, PTEN could distinguish early BC stages with good sensitivity 84.4%, 51.4%, 69.1%, 72%, and 70%, respectively. Methylation status of PTEN and SMAD4 is a promising blood marker for early detection of breast cancer. Future studies are needed for their role as prognostic markers.
Tài liệu tham khảo
Salta S, Nunes SP, Sousa MF, Lopes P, Freitas M, Caldas M, Antunes L, Castro F, Antunes p et al (2018) A DNA methylation-based test for breast cancer detection in circulating cell-free DNA. J Clin Med 7(11):420. https://doi.org/10.3390/jcm7110420
Al-thoubaity FK (2020) Molecular classification of breast cancer: a retrospective cohort study. Ann Med Surg 49:44–48. https://doi.org/10.1016/j.amsu.2019.11.021
Mahvi DA, Liu R, Grinstaff MW, Colson YL, Raut CP (2018) Local cancer recurrence: the realities, challenges, and opportunities for new therapies. CA Cancer J Clin 68(6):488–505. https://doi.org/10.3322/caac.21498
Rodgers KM, Udesky JO, Rudel RA, Brody JG (2018) Environmental chemicals and breast cancer: an updated review of epidemiological literature informed by biological mechanisms. Environ Res 160:152–182. https://doi.org/10.1016/j.envres.2017.08.045
Breast Cancer Treatment (PDQ®)(2014) Archived from the original on 5 July 2014. Retrieved 29 June 2014.
Corsetti V, Houssami N, Ghirardi M, Ferrari A, Speziani M, Bellarosa S, Remida G, Gasparotti C, Galligioni E, Ciatto S (2011) Evidence of the effect of adjunct ultrasound screening in women with mammography-negative dense breasts: interval breast cancers at 1 year follow-up. Eur J Cancer 47(7):1021–1026. https://doi.org/10.1016/j.ejca.2010.12.002
Attallah AM, El-Far M, Omran MM, Abdallah SO, El-Desouky MA, El-Dosoky I, Abdelrazek MA, Attallah AA, Elweresh MA et al (2014) Circulating levels and clinical implications of epithelial membrane antigen and cytokeratin-1 in women with breast cancer: can their ratio improve the results? Tumour Biol 35(11):10737–10745. https://doi.org/10.1007/s13277-014-2375-1
Uramova S, Kubatka P, Dankova Z, Kapinova A, Zolakova B, Samec M, Zubor P, Zulli A, Valentova V, Kwon TK, Solar P, Kello M, Kajo K, Busselberg D, Pec M, Danko J (2018) Plant natural modulators in breast cancer prevention: status quo and future perspectives reinforced by predictive, preventive, and personalized medical approach. EPMA J 9(4):403–419. https://doi.org/10.1007/s13167-018-0154-6
Zeng Y, Chen T (2019) DNA methylation reprogramming during mammalian development. Genes 10:257
Van Hoesel AQ, Sato Y, Elashoff DA, Turner RR, Giuliano AE, Shamonki JM, Kuppen PJ, van de Velde CJ, Hoon DS (2013) Assessment of DNA methylation status in early stages of breast cancer development. Br J Cancer 108(10):2033–2038. https://doi.org/10.1038/bjc.2013.136
Vidotto T, Tiezzi DN, Squire JA (2018) Distinct subtypes of genomic PT EN deletion size influence the landscape of aneuploidy and outcome in prostate cancer. Mol Cytogenet 11(1):1. https://doi.org/10.1186/s13039-017-0348-y
Reid AHM, Attard G, Ambroisine L, Fisher G, Kovacs G, Brewer D, Clark J, Flohr P, Edwards S, Berney DM, Foster CS, Fletcher A, Gerald WL, Møller H et al (2010) Molecular characterisation of ERG, ETV1 and PTEN gene loci identifies patients at lowand high risk of death from prostate cancer. Br J Cancer 102(4):678–684. https://doi.org/10.1038/sj.bjc.6605554
Rizvi MMA, Alam MS, Ali A, Mehdi SJ, Batra S, Mandal AK (2011) Abrrant promoter methylation and inactivation of PTEN gene in cervical carcinoma from Indian population. J Cancer Res Clin Oncol 137(8):1255–1262. https://doi.org/10.1007/s00432-011-0994-0
Zhang HY, Liang F, Jia ZL, Song ST, Jiang ZF (2013) PTEN mutation, methylation and expression in breast cancer patients. Oncol Lett 6(1):161–168
Lu YM, Cheng F, Teng LS (2016) The association between phosphatase and tensin homologhypermethylation and patients with breast cancer, a meta-analysis and literature review. Sci Rep 6(1):32723. https://doi.org/10.1038/srep32723
Alam MS, Jerah ABA, Ashraf AM, Kumaresan K, Eisa ZM, Mikhail NT (2017) Promoter methylation and loss of expression of PTEN gene in breast cancer patients from Saudi population. J Clin Exp Oncol 6:6
Chow JYC, Dong H, Quach KT, Van Nguyen PN, Chen K, Carethers JM (2008) TGF-β mediates PTEN suppression and cell motility through calcium-dependent PKC-α activation in pancreatic cancer cells. Am J Physiol Gastrointest Liver Physiol 294(4):899–905
Zhao M, Mishra L, Deng CX (2018) The role of TGF-β/SMAD4 signaling in cancer. Int J Biol Sci 14(2):111–123. https://doi.org/10.7150/ijbs.23230
Ullah I, Sun W, Tang L, Feng J (2018) Roles of Smads family and alternative splicing variants of Smad4 in different cancers. J Cancer 9(21):4018–4028. https://doi.org/10.7150/jca.20906
Nikolić A, Opinćal F, Ristanović M, Trifunović J, Knežević S, Radojkovic D (2017) Analysis of SMAD4 gene promoter methylation in pancreatic and endometrial cancer. Arch Oncol 23(2):17–19. https://doi.org/10.2298/AOO1702017N
Kovacevic Z, Chikhani S, Lui GYL, Sivagurunathan S, Richardson DR (2013) The iron-regulated metastasis suppressor NDRG1 targets NEDD4L, PTEN, and SMAD4 and inhibits the PI3K and Ras signaling pathways. Antioxid Redox Signal 18(8):874–887. https://doi.org/10.1089/ars.2011.4273
Chung Y, Chan Wi Y, Kim Y, Bang SS, Yang J, Jang K, Min KW, Paik SS (2018) The Smad4/PTEN expression pattern predicts clinical outcomes in colorectal adenocarcinoma. J Pathol Transl Med 52(1):37–44. https://doi.org/10.4132/jptm.2017.10.20
Greene FL, Sobin LH (2008) The staging of cancer: a retrospective and prospective appraisal. CA Cancer J Clin 58(3):180–190. https://doi.org/10.3322/CA.2008.0001
Cicco PD, Catani MV, Gasperi V, Sibilano M, Quaglietta M, Savini I (2019) Nutrition and breast cancer: a literature review on prevention, treatment and recurrence. Nutrients 11(7):1514. https://doi.org/10.3390/nu11071514
Ibrahim AS, Khaled HM, Mikhail NN, Baraka H, Kamel H (2014) Cancer incidence in Egypt: results of the national population-based cancer registry program. J Cancer Epidemiol:2014
Marina M, Dragan J, Aleksandar M, Dragan S (2017) Early diagnosis and detection of breast cancer. J Technol Health Care 26:729–759
Locke WJ, Guanzon D, Ma C, Liew Y, Duesing KR, Fung KY, Ross JP (2019) DNA methylation cancer biomarkers: translation to the clinic. Front Genet 10:1150
Xu X, Ehdaie B, Ohara N, Yoshino T, Deng CX (2010) Synergistic action of Smad4 and PTEN in suppressing pancreatic ductal adenocarcinoma formation in mice. Oncogene 29(5):674–686. https://doi.org/10.1038/onc.2009.375
Liu J, Cho SN, Akkanti B, Jin N, Mao J, Long W, Chen T, Zhang Y, Tang X, Wistub II, Creighton CJ, Kheradmand F, DeMayo FJ (2015) ErbB2 pathway activation upon Smad4 loss promotes lung tumor growth and metastasis. Cell Rep 10(9):1599–1613. https://doi.org/10.1016/j.celrep.2015.02.014
Li XB, Yang G, Zhu L, Tang YL, Zhang C, Ju Z, Yang X, Teng Y (2016) Gastric Lgr5 (+) stem cells are the cellular origin of invasive intestinal-type gastric cancer in mice. Cell Res 26(7):838–849. https://doi.org/10.1038/cr.2016.47
Syria JC, Lee HY, Lee JI, Wang L, Issa JP et al (2002) Lack of PTEN expression in non-small cell lung cancer could be related to promoter methylation. Clin Cancer Res 8:1178–1184
Kang YH, Lee HS, Kim WH (2002) Promoter methylation and silencing of PTEN in gastric carcinoma. Lab Investig 82(3):285–291. https://doi.org/10.1038/labinvest.3780422
Yang HJ, Liu SW, Wang Y, Tsang CK, Ngan YS (2006) Differential DNA methylation profiles in gynecological cancers and correlation with clinic-pathological data. BMC Cancer 6(1):212. https://doi.org/10.1186/1471-2407-6-212
Salvesen HB, MacDonald N, Ryan A, Jacobs IJ, Lynch ED, Akslen LA, Das S (2001) PTEN methylation is associated with advanced stage and microsatellite instability in endometrial carcinoma. Int J Cancer 91(1):22–26. https://doi.org/10.1002/1097-0215(20010101)91:1<22::AID-IJC1002>3.0.CO;2-S
Wu L, Shen Y, Peng X, Zhang S, Wang M, Xu G, Zheng X, Wang J, Lu C (2016) Aberrant promoter methylation of cancer-related genes in human breast cancer. Oncol Lett 12(6):5145–5155. https://doi.org/10.3892/ol.2016.5351
Yari K, Payandeh M, Rahimi Z (2015) Association of the hypermethylation status of PTEN tumor suppressor gene with the risk of breast cancer among Kurdish population from Western Iran. Int Soc Oncol BioMarkers (ISOBM)
Kazim Z, Wahabi K, Perwez A, Lal P, Rizvi MA (2019) PTEN genetic and epigenetic alterations define distinct subgroups in North Indian breast cancer patients. Asian Pac J Cancer Prev 20(1):269–276. https://doi.org/10.31557/APJCP.2019.20.1.269
Romero-Garcia S, Prado-Garcia H, Carlos-Reyes A (2020) Role of DNA methylation in the resistance to therapy in solid Tumors. Front Oncol 7(10):1152
Ali NA, McKay MJ, Molloy MP (2010) Proteomics of Smad4 regulated transforming growth factor-beta signalling in colon cancer cells. Mol BioSystems 6(11):2332–2338. https://doi.org/10.1039/c0mb00016g
Yoo SY, Lee JA, Shin Y, Cho NY, Bae JM, Kang GH (2019) Clinicopathological characterization and prognostic implication of SMAD4 expression in colorectal carcinoma. J Pathol Transl Med 53(5):289–297. https://doi.org/10.4132/jptm.2019.06.07
Wilentz R, Iacobuzio-Donahue C, Argani P, McCarthy DM, Parsons JL, Yeo CJ, Kern SE, Hruban RH (2000) Loss of expression of Dpc4 in pancreatic intraepithelial neoplasia: evidence that DPC4 inactivation occurs late in neoplastic progression. Cancer Res 60(7):2002–2006
Liu S, Foulkes WD, Leung S, Gao D, Lau S, Kos Z, Nielsen TO (2014) Prognostic significance of FOXP3+ tumor-infiltrating lymphocytes in breast cancer depends on estrogen receptor and human epidermal growth factor receptor-2 expression status and concurrent cytotoxic T-cell infiltration. Breast Cancer Res 16(5):432. https://doi.org/10.1186/s13058-014-0432-8
Deckers M, van Dinther M, Buijs J, Que I, Löwik C, van der Pluijm G, ten Dijke P (2006) The tumor suppressor Smad4 is required for transforming growth factor beta-induced epithelial to mesenchymal transition and bone metastasis of breast cancer cells. Cancer Res 66(4):2202–2209. https://doi.org/10.1158/0008-5472.CAN-05-3560
Li Q, Wu L, Oelschlager DK, Wan M, Stockard CR, Grizzle WE, Wang N, Chen H, Sun Y, Cao X (2005) Smad4 inhibits tumor growth by inducing apoptosis in estrogen receptor-alpha-positive breast cancer cells. J Biol Chem 280(29):27022–27028. https://doi.org/10.1074/jbc.M505071200
Roth S, Laiho P, Salovaara R, Launonen V, Aaltonen LA (2000) No SMAD4 hypermethylation in colorectal cancer. Br J Cancer 83(8):1015–1019. https://doi.org/10.1054/bjoc.2000.1387
Sameer AS, Siddiqi MA (2011) SMAD4 promoter hypermethylation in Kashmiri colorectal cancer cases. Saudi J Gastroenterol 17(4):297. https://doi.org/10.4103/1319-3767.82591
Fotouhi O, Adel Fahmideh M, Kjellman M, Sulaiman L, Hoog A, Zedenius J, Hashemi J, Larsson C (2014) Global hypomethylation and promoter methylation in small intestinal neuroendocrine tumors: an in vivo and in vitro study. Epigenetics 9(7):987–997. https://doi.org/10.4161/epi.28936
Onwuegbusi BA, Aitchison A, Chin SF, Kranjac T, Mills I, Huang Y, Lao-Sirieix P, Caldas C, Fitzgerald RC (2006) Impaired transforming growth factor beta signalling in Barrett’s carcinogenesis due to frequent SMAD4 inactivation. Gut 55(6):764–774. https://doi.org/10.1136/gut.2005.076430