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Erschienen in: Cellular Oncology 4/2014

01.08.2014 | Original Paper

Validation of DNA promoter hypermethylation biomarkers in breast cancer — a short report

verfasst von: Jolien S. de Groot, Xiaojuan Pan, Jan Meeldijk, Elsken van der Wall, Paul J. van Diest, Cathy B. Moelans

Erschienen in: Cellular Oncology | Ausgabe 4/2014

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Abstract

Purpose

DNA promoter hypermethylation of tumor suppressor genes is known to occur early in cancer development, including breast cancer. To improve early breast cancer detection, we aimed to investigate whether the identification of DNA promoter hypermethylation might be of added value.

Methods

The methylation status of a panel of 19 candidate genes (AKR1B1, ALX1, ARHGEF7, FZD10, GHSR, GPX7, GREM1, GSTP1, HOXD1, KL, LHX2, MAL, MGMT, NDRG2, RASGRF2, SFRP1, SFRP2, TM6SF1 and TMEFF2) was determined in formalin-fixed paraffin-embedded normal breast and breast cancer tissue samples using gel-based methylation-specific PCR (MSP).

Results

The promoters of the AKR1B1, ALX1, GHSR, GREM1, RASGRF2, SFRP2, TM6SF1 and TMEFF2 genes were found to be significantly differentially methylated in normal versus malignant breast tissues. Based on sensitivity, specificity and logistic regression analyses the best performing genes for detecting breast cancer were identified. Through multivariate analyses, we found that AKR1B1 and TM6SF1 could detect breast cancer with an area under the curve (AUC) of 0.986 in a receiver operating characteristic (ROC) assessment.

Conclusions

Based on our data, we conclude that AKR1B1 and TM6SF1 may serve as candidate methylation biomarkers for early breast cancer detection. Further studies are underway to evaluate the methylation status of these genes in body fluids, including nipple aspirates and blood.
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Literatur
2.
Zurück zum Zitat A. Algamas-Dimantov, E. Yehuda-Shnaidman, I. Peri, B. Schwartz, Epigenetic control of HNF-4alpha in colon carcinoma cells affects MUC4 expression and malignancy. Cell. Oncol. 36, 155–167 (2013)CrossRef A. Algamas-Dimantov, E. Yehuda-Shnaidman, I. Peri, B. Schwartz, Epigenetic control of HNF-4alpha in colon carcinoma cells affects MUC4 expression and malignancy. Cell. Oncol. 36, 155–167 (2013)CrossRef
3.
Zurück zum Zitat Y. You, W. Yang, Z. Wang, H. Zhu, H. Li, C. Lin, Y. Ran, Promoter hypermethylation contributes to the frequent suppression of the CDK10 gene in human nasopharyngeal carcinomas. Cell. Oncol. 36, 323–331 (2013)CrossRef Y. You, W. Yang, Z. Wang, H. Zhu, H. Li, C. Lin, Y. Ran, Promoter hypermethylation contributes to the frequent suppression of the CDK10 gene in human nasopharyngeal carcinomas. Cell. Oncol. 36, 323–331 (2013)CrossRef
4.
Zurück zum Zitat L.H. Jensen, A.A. Rasmussen, L. Byriel, H. Kuramochi, D.G. Cruger, J. Lindebjerg, P.V. Danenberg, A. Jakobsen, K. Danenberg, Regulation of MLH1 mRNA and protein expression by promoter methylation in primary colorectal cancer: A descriptive and prognostic cancer marker study. Cell. Oncol. 36, 411–419 (2013)CrossRef L.H. Jensen, A.A. Rasmussen, L. Byriel, H. Kuramochi, D.G. Cruger, J. Lindebjerg, P.V. Danenberg, A. Jakobsen, K. Danenberg, Regulation of MLH1 mRNA and protein expression by promoter methylation in primary colorectal cancer: A descriptive and prognostic cancer marker study. Cell. Oncol. 36, 411–419 (2013)CrossRef
5.
Zurück zum Zitat A. Geurts van Kessel, The cancer genome: From structure to function. Cell. Oncol. 37, 155–165 (2014)CrossRef A. Geurts van Kessel, The cancer genome: From structure to function. Cell. Oncol. 37, 155–165 (2014)CrossRef
6.
Zurück zum Zitat K.P. Suijkerbuijk, M.J. Fackler, S. Sukumar, C.H. van Gils, T. van Laar, E. van der Wall, M. Vooijs, P.J. van Diest, Methylation is less abundant in BRCA1-associated compared with sporadic breast cancer. Ann. Oncol. 19, 1870–1874 (2008)PubMedCentralPubMedCrossRef K.P. Suijkerbuijk, M.J. Fackler, S. Sukumar, C.H. van Gils, T. van Laar, E. van der Wall, M. Vooijs, P.J. van Diest, Methylation is less abundant in BRCA1-associated compared with sporadic breast cancer. Ann. Oncol. 19, 1870–1874 (2008)PubMedCentralPubMedCrossRef
7.
Zurück zum Zitat J.G. Herman, J.R. Graff, S. Myohanen, B.D. Nelkin, S.B. Baylin, Methylation-specific PCR: A novel PCR assay for methylation status of CpG islands. Proc. Natl. Acad. Sci. U. S. A. 93, 9821–9826 (1996)PubMedCentralPubMedCrossRef J.G. Herman, J.R. Graff, S. Myohanen, B.D. Nelkin, S.B. Baylin, Methylation-specific PCR: A novel PCR assay for methylation status of CpG islands. Proc. Natl. Acad. Sci. U. S. A. 93, 9821–9826 (1996)PubMedCentralPubMedCrossRef
8.
Zurück zum Zitat D.S. Shames, J.D. Minna, A.F. Gazdar, Methods for detecting DNA methylation in tumors: From bench to bedside. Cancer Lett. 251, 187–198 (2007)PubMedCrossRef D.S. Shames, J.D. Minna, A.F. Gazdar, Methods for detecting DNA methylation in tumors: From bench to bedside. Cancer Lett. 251, 187–198 (2007)PubMedCrossRef
9.
Zurück zum Zitat P.J. van Diest, No consent should be needed for using leftover body material for scientific purposes. BMJ 325, 648–651 (2002)PubMedCrossRef P.J. van Diest, No consent should be needed for using leftover body material for scientific purposes. BMJ 325, 648–651 (2002)PubMedCrossRef
10.
Zurück zum Zitat T. Swift-Scanlan, R. Vang, A. Blackford, M.J. Fackler, S. Sukumar, Methylated genes in breast cancer: Associations with clinical and histopathological features in a familial breast cancer cohort. Cancer Biol. Ther. 11, 853–865 (2011)PubMedCentralPubMedCrossRef T. Swift-Scanlan, R. Vang, A. Blackford, M.J. Fackler, S. Sukumar, Methylated genes in breast cancer: Associations with clinical and histopathological features in a familial breast cancer cohort. Cancer Biol. Ther. 11, 853–865 (2011)PubMedCentralPubMedCrossRef
11.
Zurück zum Zitat M.J. Fackler, C.B. Umbricht, D. Williams, P. Argani, L.A. Cruz, V.F. Merino, W.W. Teo, Z. Zhang, P. Huang, K. Visvananthan, J. Marks, S. Ethier, J.W. Gray, A.C. Wolff, L.M. Cope, S. Sukumar, Genome-wide methylation analysis identifies genes specific to breast cancer hormone receptor status and risk of recurrence. Cancer Res. 71, 6195–6207 (2011)PubMedCentralPubMedCrossRef M.J. Fackler, C.B. Umbricht, D. Williams, P. Argani, L.A. Cruz, V.F. Merino, W.W. Teo, Z. Zhang, P. Huang, K. Visvananthan, J. Marks, S. Ethier, J.W. Gray, A.C. Wolff, L.M. Cope, S. Sukumar, Genome-wide methylation analysis identifies genes specific to breast cancer hormone receptor status and risk of recurrence. Cancer Res. 71, 6195–6207 (2011)PubMedCentralPubMedCrossRef
12.
Zurück zum Zitat M.J. Fackler, Z.L. Bujanda, C. Umbricht, W.W. Teo, S. Cho, Z. Zhang, K. Visvanathan, S. Jeter, P. Argani, C. Wang, J.P. Lyman, M. de Brot, J.N. Ingle, J. Boughey, K. McGuire, T.A. King, L.A. Carey, L. Cope, A.C. Wolff, S. Sukumar, Novel methylated biomarkers and a robust assay to detect circulating tumor DNA in metastatic breast cancer. Cancer Res. 74, 2160–2170 (2014)PubMedCrossRef M.J. Fackler, Z.L. Bujanda, C. Umbricht, W.W. Teo, S. Cho, Z. Zhang, K. Visvanathan, S. Jeter, P. Argani, C. Wang, J.P. Lyman, M. de Brot, J.N. Ingle, J. Boughey, K. McGuire, T.A. King, L.A. Carey, L. Cope, A.C. Wolff, S. Sukumar, Novel methylated biomarkers and a robust assay to detect circulating tumor DNA in metastatic breast cancer. Cancer Res. 74, 2160–2170 (2014)PubMedCrossRef
14.
15.
Zurück zum Zitat N. Hevir, J. Sinkovec, T. Lanisnik Rizner, Decreased levels of AKR1B1 and AKR1B10 in cancerous endometrium compared to adjacent non-cancerous tissue. Chem. Biol. Interact. 202, 226–233 (2013)PubMedCrossRef N. Hevir, J. Sinkovec, T. Lanisnik Rizner, Decreased levels of AKR1B1 and AKR1B10 in cancerous endometrium compared to adjacent non-cancerous tissue. Chem. Biol. Interact. 202, 226–233 (2013)PubMedCrossRef
16.
Zurück zum Zitat B. Laffin, J.M. Petrash, Expression of the Aldo-Ketoreductases AKR1B1 and AKR1B10 in Human Cancers. Front. Pharmacol. 3, 104 (2012)PubMedCentralPubMed B. Laffin, J.M. Petrash, Expression of the Aldo-Ketoreductases AKR1B1 and AKR1B10 in Human Cancers. Front. Pharmacol. 3, 104 (2012)PubMedCentralPubMed
17.
Zurück zum Zitat R. Tao, J. Li, J. Xin, J. Wu, J. Guo, L. Zhang, L. Jiang, W. Zhang, Z. Yang, L. Li, Methylation profile of single hepatocytes derived from hepatitis B virus-related hepatocellular carcinoma. PLoS One 6, e19862 (2011)PubMedCentralPubMedCrossRef R. Tao, J. Li, J. Xin, J. Wu, J. Guo, L. Zhang, L. Jiang, W. Zhang, Z. Yang, L. Li, Methylation profile of single hepatocytes derived from hepatitis B virus-related hepatocellular carcinoma. PLoS One 6, e19862 (2011)PubMedCentralPubMedCrossRef
18.
Zurück zum Zitat F. Fang, S. Turcan, A. Rimner, A. Kaufman, D. Giri, L.G. Morris, R. Shen, V. Seshan, Q. Mo, A. Heguy, S.B. Baylin, N. Ahuja, A. Viale, J. Massaque, L. Norton, L.T. Vahdat, M.E. Moynahan, T.A. Chan, Breast cancer methylomes establish an epigenomic foundation for metastasis. Sci. Transl. Med. 3, 75ra25 (2011)PubMedCentralPubMed F. Fang, S. Turcan, A. Rimner, A. Kaufman, D. Giri, L.G. Morris, R. Shen, V. Seshan, Q. Mo, A. Heguy, S.B. Baylin, N. Ahuja, A. Viale, J. Massaque, L. Norton, L.T. Vahdat, M.E. Moynahan, T.A. Chan, Breast cancer methylomes establish an epigenomic foundation for metastasis. Sci. Transl. Med. 3, 75ra25 (2011)PubMedCentralPubMed
19.
Zurück zum Zitat R.T. Chatterton, M. Sahadevan, O. Lee, H. Hong, W. Jun, I. Helenowski, S. Sukumar, V. Stearns, M.J. Fackler, S.A. Khan, Single-strand breaks in normal breast DNA are associated with measures of breast cancer risk, deficiencies in repair mechanisms, and diminished protection against oxidative processes. Cancer Prev. Res. 5, A15 (2012)CrossRef R.T. Chatterton, M. Sahadevan, O. Lee, H. Hong, W. Jun, I. Helenowski, S. Sukumar, V. Stearns, M.J. Fackler, S.A. Khan, Single-strand breaks in normal breast DNA are associated with measures of breast cancer risk, deficiencies in repair mechanisms, and diminished protection against oxidative processes. Cancer Prev. Res. 5, A15 (2012)CrossRef
20.
Zurück zum Zitat S.K. Botla, A.M. Gholami, M. Malekpour, E.A. Moskalev, M. Fallah, P. Jandaghi, A. Aghajani, I.S. Bondar, R. Omranipour, F. Malekpour, A. Mohajeri, A.J. Bababi, O. Sahin, V.V. Bubnov, H. Najmabadi, J.D. Hoheisel, Y. Riazalhosseini, Diagnostic values of GHSR DNA methylation pattern in breast cancer. Breast Cancer Res. Treat. 135, 705–713 (2012)PubMedCrossRef S.K. Botla, A.M. Gholami, M. Malekpour, E.A. Moskalev, M. Fallah, P. Jandaghi, A. Aghajani, I.S. Bondar, R. Omranipour, F. Malekpour, A. Mohajeri, A.J. Bababi, O. Sahin, V.V. Bubnov, H. Najmabadi, J.D. Hoheisel, Y. Riazalhosseini, Diagnostic values of GHSR DNA methylation pattern in breast cancer. Breast Cancer Res. Treat. 135, 705–713 (2012)PubMedCrossRef
21.
Zurück zum Zitat J. Jeschke, L. van Neste, S.C. Glockner, M. Dhir, M.F. Calmon, V. Deregowski, W. van Criekinge, I. Vlassenbroeck, A. Koch, T.A. Chan, L. Cope, C.M. Hooker, K.E. Schuebel, E. Gabrielson, A. Winterpacht, S.B. Baylin, J.G. Herman, N. Ahuja, Biomarkers for detection and prognosis of breast cancer identified by a functional hypermethylome screen. Epigenetics 7, 701–709 (2012)PubMedCrossRef J. Jeschke, L. van Neste, S.C. Glockner, M. Dhir, M.F. Calmon, V. Deregowski, W. van Criekinge, I. Vlassenbroeck, A. Koch, T.A. Chan, L. Cope, C.M. Hooker, K.E. Schuebel, E. Gabrielson, A. Winterpacht, S.B. Baylin, J.G. Herman, N. Ahuja, Biomarkers for detection and prognosis of breast cancer identified by a functional hypermethylome screen. Epigenetics 7, 701–709 (2012)PubMedCrossRef
22.
Zurück zum Zitat S.Y. Park, H.J. Kwon, H.E. Lee, H.S. Ryu, S.W. Kim, J.H. Kim, I.A. Kim, N. Jung, N.Y. Cho, G.H. Kang, Promoter CpG island hypermethylation during breast cancer progression. Virchows Arch. 458, 73–84 (2011)PubMedCrossRef S.Y. Park, H.J. Kwon, H.E. Lee, H.S. Ryu, S.W. Kim, J.H. Kim, I.A. Kim, N. Jung, N.Y. Cho, G.H. Kang, Promoter CpG island hypermethylation during breast cancer progression. Virchows Arch. 458, 73–84 (2011)PubMedCrossRef
23.
Zurück zum Zitat R. Krassenstein, E. Sauter, E. Dulaimi, C. Battagli, H. Ehya, A. Klein-Szanto, P. Cairns, Detection of breast cancer in nipple aspirate fluid by CpG island hypermethylation. Clin. Cancer Res. 10, 28–32 (2004)PubMedCrossRef R. Krassenstein, E. Sauter, E. Dulaimi, C. Battagli, H. Ehya, A. Klein-Szanto, P. Cairns, Detection of breast cancer in nipple aspirate fluid by CpG island hypermethylation. Clin. Cancer Res. 10, 28–32 (2004)PubMedCrossRef
24.
Zurück zum Zitat M. Faryna, C. Konermann, S. Aulmann, J.L. Bermejo, M. Brugger, S. Diederichs, J. Rom, D. Weichenhan, R. Claus, M. Rehli, P. Schirmacher, H.P. Sinn, C. Plass, C. Gerhauser, Genome-wide methylation screen in low-grade breast cancer identifies novel epigenetically altered genes as potential biomarkers for tumor diagnosis. FASEB J. 26, 4937–4950 (2012)PubMedCrossRef M. Faryna, C. Konermann, S. Aulmann, J.L. Bermejo, M. Brugger, S. Diederichs, J. Rom, D. Weichenhan, R. Claus, M. Rehli, P. Schirmacher, H.P. Sinn, C. Plass, C. Gerhauser, Genome-wide methylation screen in low-grade breast cancer identifies novel epigenetically altered genes as potential biomarkers for tumor diagnosis. FASEB J. 26, 4937–4950 (2012)PubMedCrossRef
25.
Zurück zum Zitat M.S. Kim, J. Lee, T. Oh, Y. Moon, E. Chang, K.S. Seo, B.D. Hoehn, S. An, J.H. Lee, Genome-wide identification of OTP gene as a novel methylation marker of breast cancer. Oncol. Rep. 27, 1681–1688 (2012)PubMed M.S. Kim, J. Lee, T. Oh, Y. Moon, E. Chang, K.S. Seo, B.D. Hoehn, S. An, J.H. Lee, Genome-wide identification of OTP gene as a novel methylation marker of breast cancer. Oncol. Rep. 27, 1681–1688 (2012)PubMed
26.
Zurück zum Zitat H.N. Horne, P.S. Lee, S.K. Murphy, M.A. Alonso, J.A. Olson, J.R. Marks, Inactivation of the MAL gene in breast cancer is a common event that predicts benefit from adjuvant chemotherapy. Mol. Cancer Res. 7, 199–209 (2009)PubMedCentralPubMedCrossRef H.N. Horne, P.S. Lee, S.K. Murphy, M.A. Alonso, J.A. Olson, J.R. Marks, Inactivation of the MAL gene in breast cancer is a common event that predicts benefit from adjuvant chemotherapy. Mol. Cancer Res. 7, 199–209 (2009)PubMedCentralPubMedCrossRef
27.
Zurück zum Zitat C. Fumagalli, G. Pruneri, P. Possanzini, M. Manzotti, M. Barile, I. Feroce, M. Colleoni, B. Bonnani, P. Maisonneuve, P. Radice, G. Viale, M. Barberis, Methylation of O6-methylguanine-DNA methyltransferase (MGMT) promoter gene in triple-negative breast cancer patients. Breast Cancer Res. Treat. 134, 131–137 (2012)PubMedCrossRef C. Fumagalli, G. Pruneri, P. Possanzini, M. Manzotti, M. Barile, I. Feroce, M. Colleoni, B. Bonnani, P. Maisonneuve, P. Radice, G. Viale, M. Barberis, Methylation of O6-methylguanine-DNA methyltransferase (MGMT) promoter gene in triple-negative breast cancer patients. Breast Cancer Res. Treat. 134, 131–137 (2012)PubMedCrossRef
28.
Zurück zum Zitat J. Klajic, T. Fleischer, E. Dejeux, H. Edvardsen, F. Warnberg, I. Bukholm, P.E. Lonning, H. Solvang, A.L. Borresen-Dale, J. Tost, V.N. Kristensen, Quantitative DNA methylation analyses reveal stage dependent DNA methylation and association to clinico-pathological factors in breast tumors. BMC Cancer 13, 1–12 (2013)CrossRef J. Klajic, T. Fleischer, E. Dejeux, H. Edvardsen, F. Warnberg, I. Bukholm, P.E. Lonning, H. Solvang, A.L. Borresen-Dale, J. Tost, V.N. Kristensen, Quantitative DNA methylation analyses reveal stage dependent DNA methylation and association to clinico-pathological factors in breast tumors. BMC Cancer 13, 1–12 (2013)CrossRef
29.
Zurück zum Zitat Y. Alkam, H. Mitomi, K. Nakai, T. Himuro, T. Saito, M. Takahashi, A. Arakawa, T. Yao, M. Saito, Protein expression and methylation of DNA repair genes hMLH1, hMSH2, MGMT and BRCA1 and their correlation with clinicopathological parameters and prognosis in basal-like breast cancer. Histopathology 63, 713–725 (2013)PubMed Y. Alkam, H. Mitomi, K. Nakai, T. Himuro, T. Saito, M. Takahashi, A. Arakawa, T. Yao, M. Saito, Protein expression and methylation of DNA repair genes hMLH1, hMSH2, MGMT and BRCA1 and their correlation with clinicopathological parameters and prognosis in basal-like breast cancer. Histopathology 63, 713–725 (2013)PubMed
30.
Zurück zum Zitat A. Tserga, N.V. Michalopoulos, G. Levidou, P. Korkolopoulou, G. Zografos, E. Patsouris, A.A. Saetta, Association of aberrant DNA methylation with clinicopathological features in breast cancer. Oncol. Rep. 27, 1630–1638 (2012)PubMed A. Tserga, N.V. Michalopoulos, G. Levidou, P. Korkolopoulou, G. Zografos, E. Patsouris, A.A. Saetta, Association of aberrant DNA methylation with clinicopathological features in breast cancer. Oncol. Rep. 27, 1630–1638 (2012)PubMed
31.
Zurück zum Zitat G. Sharma, S. Mirza, R. Parshad, A. Srivastava, S.D. Gupta, P. Pandya, R. Ralhan, Clinical significance of promoter hypermethylation of DNA repair genes in tumor and serum DNA in invasive ductal breast carcinoma patients. Life Sci. 87, 83–91 (2010)PubMedCrossRef G. Sharma, S. Mirza, R. Parshad, A. Srivastava, S.D. Gupta, P. Pandya, R. Ralhan, Clinical significance of promoter hypermethylation of DNA repair genes in tumor and serum DNA in invasive ductal breast carcinoma patients. Life Sci. 87, 83–91 (2010)PubMedCrossRef
32.
Zurück zum Zitat A.A. Muggerud, J.A. Ronneberg, F. Warnberg, J. Botling, F. Busato, J. Jovanovic, H. Solvang, I. Bukholm, A.L. Borresen-Dale, V.N. Kristensen, T. Sorlie, J. Tost, Frequent aberrant DNA methylation of ABCB1, FOXC1, PPP2R2B and PTEN in ductal carcinoma in situ and early invasive breast cancer. Breast Cancer Res. 12, R3 (2010)PubMedCentralPubMedCrossRef A.A. Muggerud, J.A. Ronneberg, F. Warnberg, J. Botling, F. Busato, J. Jovanovic, H. Solvang, I. Bukholm, A.L. Borresen-Dale, V.N. Kristensen, T. Sorlie, J. Tost, Frequent aberrant DNA methylation of ABCB1, FOXC1, PPP2R2B and PTEN in ductal carcinoma in situ and early invasive breast cancer. Breast Cancer Res. 12, R3 (2010)PubMedCentralPubMedCrossRef
Metadaten
Titel
Validation of DNA promoter hypermethylation biomarkers in breast cancer — a short report
verfasst von
Jolien S. de Groot
Xiaojuan Pan
Jan Meeldijk
Elsken van der Wall
Paul J. van Diest
Cathy B. Moelans
Publikationsdatum
01.08.2014
Verlag
Springer Netherlands
Erschienen in
Cellular Oncology / Ausgabe 4/2014
Print ISSN: 2211-3428
Elektronische ISSN: 2211-3436
DOI
https://doi.org/10.1007/s13402-014-0189-1

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