Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
BMC Cancer
Published in: BMC Cancer 1/2010

Open Access 01-12-2010 | Research article

Genetic and epigenetic silencing of the beclin 1gene in sporadic breast tumors

Authors: Zidong Li, Bo Chen, Yiqing Wu, Feng Jin, Yongjing Xia, Xiangjun Liu

Published in: BMC Cancer | Issue 1/2010

Login to get access

Abstract

Background

Beclin 1, an important autophagy-related protein in human cells, is involved in cell death and cell survival. Beclin 1 mapped to human chromosome 17q21. It is widely expressed in normal mammary epithelial cells. Although down-regulated expression with mono-allelic deletions of beclin 1 gene was frequently observed in breast tumors, whether there was other regulatory mechanism of beclin 1 was to be investigated. We studied the expression of beclin 1 and explored the possible regulatory mechanisms on its expression in breast tumors.

Methods

20 pairs of tumors and adjacent normal tissues from patients with sporadic breast invasive ductal cancer (IDCs) were collected. The mRNA expression of beclin 1 was detected by real-time quantitative RT-PCR. Loss of heterozygosity (LOH) was determined by real-time quantitative PCR and microsatellite methods. The protein expression of beclin 1, p53, BRCA1 and BRCA2 was assessed by immunohistochemistry. CpG islands in 5' genomic region of beclin 1 gene were identified using MethylPrimer Program. Sodium bisulfite sequencing was used in examining the methylation status of each CpG island.

Results

Decreased beclin 1 mRNA expression was detected in 70% of the breast tumors, and the protein levels were co-related to the mRNA levels. Expression of beclin 1 mRNA was demonstrated to be much higher in the BRCA1 positive tumors than that in the BRCA1 negative ones. Loss of heterozygosity was detected in more than 45% of the breast tumors, and a dense cluster of CpG islands was found from the 5' end to the intron 2 of the beclin 1 gene. Methylation analysis showed that the promoter and the intron 2 of beclin 1 were aberrantly methylated in the tumors with decreased expression.

Conclusions

These data indicated that LOH and aberrant DNA methylation might be the possible reasons of the decreased expression of beclin 1 in the breast tumors. The findings here shed some new light on the regulatory mechanisms of beclin 1 in breast cancer.
Appendix
Available only for authorised users
Literature
1.
Edinger AL, Thompson CB: Defective autophagy leads to cance. Cancer Cell. 2003, 4: 422-424. 10.1016/S1535-6108(03)00306-4.CrossRefPubMed
2.
3.
Liang XH, Jaskon S, Seaman M, Brown K, Kempkes B, Hibshoosh H, Levine B: Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature. 1999, 402: 672-676. 10.1038/45257.CrossRefPubMed
4.
Yue Z, Jin S, Yang C, Yang C, Levine B: Beclin 1, an autophagy gene essential for early embryonic development, is a haploinsufficient tumor suppressor. Proc Natl Acad Sci. 2003, 100: 15077-15082. 10.1073/pnas.2436255100.CrossRefPubMedPubMedCentral
5.
Liang XH, Kleeman LK, Jiang HH, Gordon G, Goldman JE, Berry G, Herman B, Levine B: Protection against fatal Sindbis virus encephalitis by Beclin, a novel Bcl-2 interacting protein. J Virol. 1998, 72: 8586-8596.PubMedPubMedCentral
6.
Liang XH, Yu J, Brown K, Levine B: Beclin 1 contains a leucine-rich nuclear export signal that is required for its autophagy and tumor suppressor function. Cancer Res. 2001, 61: 3443-3449.PubMed
7.
Furuya D, Tsuji N, Yagihashi A, Watanabe N: Beclin 1 augmented cis-diamminedichloroplatinum induced apoptosis via enhancing caspase-9 activity. Exp Cell Res. 2005, 307: 26-40. 10.1016/j.yexcr.2005.02.023.CrossRefPubMed
8.
Daniel F, Legrand A, Pessayre D, Vadrot N, Descatoire V, Bernuau D: Partial Beclin 1 silencing aggravates doxorubicin- and Fas-induced apoptosis in HepG2 cells. World J Gastroenterol. 2006, 12: 2895-2900.CrossRefPubMedPubMedCentral
9.
Aita VM, Liang XH, Murty VV, Pincus DL, Yu WP, Cayanis E, Kalachikov S, Gilliam TC, Levine B: Cloning and genomic organization of beclin 1, a candidate tumor suppressor gene on chromosome 17q21. Genomics. 1999, 59: 59-65. 10.1006/geno.1999.5851.CrossRefPubMed
10.
Russell SE, Hickey GI, Lowry WS, White P, Atkinson RJ: Allele loss from chromosome 17 in ovarian cance. Oncogene. 1990, 7: 2069-2072.
11.
Sato T, Saito H, Morita R, Koi S, Lee JH, Nakamura Y: Allelotype of human ovarian cancer. Cancer Res. 1991, 51: 5118-5122.PubMed
12.
Futreal PA, Soderkvist P, Marks JR, Iglehart JD, Cochran C, Barrett JC, Wiseman RW: Detection of frequent allelic loss on proximal chromosome 17q in sporadic breast carcinoma using microsatellite length polymorphisms. Cancer Res. 1992, 52: 2624-2627.PubMed
13.
Gao X, Zacharek A, Salkowski A, Grignon DJ, Sakr W, Porter AT, Honn KV: Loss of heterozygosity of the BRCA1 and other loci on chromosome 17q in human prostate cancer. Cancer Res. 1995, 55: 1002-1005.PubMed
14.
Miremadi A, Oestergaard MZ, Paul PD, Caldas C: Cancer genetics of epigenetic genes. Hum Mol Genet. 2007, 16: 28-49. 10.1093/hmg/ddm021.CrossRef
15.
Widschwendter M, Jones PA: The potential prognostic, predictive, and therapeutic values of DNA methylation in cancer. Clin Cancer Res. 2002, 8: 17-21.PubMed
16.
Esteller M, Corn PG, Baylin SB, Herman JG: A gene hypermethylation profile of human cancer. Cancer Res. 2001, 61: 3225-3229.PubMed
17.
Weinmann AS, Bartley SM, Zhang T, Zhang MQ, Farnham PJ: Use of chromatin immunoprecipitation to clone novel E2F target promoters. Mol Cell Biol. 2001, 21: 6820-6832. 10.1128/MCB.21.20.6820-6832.2001.CrossRefPubMedPubMedCentral
18.
Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods. 2001, 25: 402-408. 10.1006/meth.2001.1262.CrossRefPubMed
19.
Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB: Methylation-specific PCR: A novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci. 1996, 93: 9821-9826. 10.1073/pnas.93.18.9821.CrossRefPubMedPubMedCentral
20.
Roessler M, Rollinger W, Mantovani-Endl L, Roessler M, Rollinger W, Mantovani-Endl L, Hagmann ML, Palme S, Berndt P, Engel AM, Pfeffer M, Karl J, Bodenmüller H, Rüschoff J, Henkel T, Rohr G, Rossol S, Rösch W, Langen H, Zolg W, Tacke M: Identification of PSME3 as a Novel Serum Tumor Marker for Colorectal Cancer by Combining Two-dimensional Polyacrylamide Gel Electrophoresis with a Strictly Mass Spectrometry-based Approach for Data Analysis. Mol Cell Proteomics. 2006, 5: 2092-2101. 10.1074/mcp.M600118-MCP200.CrossRefPubMed
21.
Zhang Q, Mashima Y, Noda S, Y Imamura, Kudoh J, Shimizu N, Nishiyama T, Umeda S, Oguchi Y, Tanaka Y, Iwata T: Characterization of AOC2 gene encoding a copper-binding amine oxidase expressed specifically in retina. Gene. 2003, 318: 45-53. 10.1016/S0378-1119(03)00753-4.CrossRefPubMed
22.
Wilson FH, Kahle KT, Sabath E, Lalioti MD, Rapson AK, Hoover RS, Hebert SC, Gamba G, Lifton RP: Molecular pathogenesis of inherited hypertension with hyperkalemia: The Na-Cl cotransporter is inhibited by wild-type but not mutant WNK4. Proc Natl Acad Sci USA. 2003, 100: 680-684. 10.1073/pnas.242735399.CrossRefPubMedPubMedCentral
23.
Feltus FA, Lee EK, Costello JF, Plass C, Vertino PM: Predicting aberrant CpG island methylation. Proc Natl Acad Sci. 2003, 100: 12253-12258. 10.1073/pnas.2037852100.CrossRefPubMedPubMedCentral
24.
Ushijima T: Detection and interpretation of altered methylation patterns in cancer cells. Nat Rev Cancer. 2005, 5: 223-231. 10.1038/nrc1571.CrossRefPubMed
25.
O'Connell P, Pekkel V, Fuqua SA, Osborne CK, Clark GM, Allred DC: Analysis of Loss of Heterozygosity in 399 Premalignant Breast Lesions at 15 Genetic Loci. J Natl Cancer Inst. 1998, 90: 697-703. 10.1093/jnci/90.9.697.CrossRefPubMed
26.
Dobrovic A, Simpfendorfer D: Methylation of the BRCA1 Gene in Sporadic Breast Cancer. Cancer Res. 1997, 57: 3347-3350.PubMed
27.
Wei M, Grushko TA, Dignam J, Hagos F, Nanda R, Sveen L, Xu JH, Fackenthal J, Tretiakova M, Das S, Olopade OI: BRCA1 promoter methylation in sporadic breast cancer is associated with reduced BRCA1 copy number and chromosome 17 aneusomy. Cancer Res. 2005, 65: 10692-10699. 10.1158/0008-5472.CAN-05-1277.CrossRefPubMed
28.
Rice JC, Massey-Brown KS, Futscher BW: Aberrant methylation of the BRCA1 CpG island promoter is associated with decreased BRCA1 mRNA in sporadic breast cancer cells. Oncogene. 1998, 17: 1807-1812. 10.1038/sj.onc.1202086.CrossRefPubMed
29.
Mueller CR, Roskelley CD: Regulation of BRCA1 expression and its relationship to sporadic breast cancer. Breast Cancer Res. 2002, 5: 45-52. 10.1186/bcr557.CrossRefPubMedPubMedCentral
30.
Coles C, Condie A, Chetty U, Steel M, Evans HJ, Prosser J: P53 mutations in breast cancer. Cancer Res. 1992, 52: 5291-5298.PubMed
31.
Greenblatt MS, Chappuis PO, Bond JP, Hamel N, Foulkes WD: TP53 mutations in breast cancer associated with BRCA1 or BRCA2 germ-line mutations: distinctive spectrum and structural distribution. Cancer Res. 2001, 61: 4092-4097.PubMed
32.
Lakhani SR, Jacquemier J, Sloane JP, Gusterson BA, Anderson TJ, Vijver MJ, Farid LM, Venter D, Antoniou A, Storfer-Isser A, Smyth E, Steel CM, Haites N, Scott RJ, Goldgar D, Neuhausen S, Daly PA, Ormiston W, Mcmanus R, Scherneck S, Ponder BA, Ford D, Peto J, Stoppa-Lyonnet D, Easton DF: Multifactorial analysis of differences between sporadic breast cancers and cancers involving BRCA1 and BRCA2 mutations. J Natl Cancer Inst. 1998, 90: 1138-1145. 10.1093/jnci/90.15.1138.CrossRefPubMed
33.
Lancaster JM, Wooster R, Mangion J, Phelan CM, Cochran C, Gumbs C, Seal S, Barfoot R, Collins N, Bignell G, Patel S, Hamoudi R, Larsson C, Wiseman RW, Berchuck A, Iglehart JD, Marks JR, Ashworth A, Stratton MR, Futreal A: BRCA2 mutations in primary breast and ovarian cancers. Nat Genet. 1996, 13: 238-240. 10.1038/ng0696-238.CrossRefPubMed
34.
Thor AD, Moore DH, Edgerton SM, Kawasaki ES, Reihsaus E, Lynch HT, Marcus JN, Schwartz L, Chen LC, Mayall BH, Smith HS: Accumulation of p53 Tumor Suppressor Gene Protein: An independent marker of prognosis in breast cancers. J Natl Cancer Inst. 1992, 84: 845-855. 10.1093/jnci/84.11.845.CrossRefPubMed
35.
Hedenfalk I, Duggan D, Chen YC, Radmacher M, Bittner M, Simon R, Meltzer P, Gusterson B, Esteller M, Kallioniemi OP, Wilfond B, Borg A, Trent J: Gene-expression profiles in hereditary breast cancer. N Engl J Med. 2001, 344: 344-539. 10.1056/NEJM200102223440801.CrossRef
36.
Wilson CA, Ramos L, Villasenor MR, Anders KH, Press MF, Clarke K, Karlan B, Chen JJ, Scully R, Livingston D, Zuch RH, Kanter MH, Cohen S, Calzone FJ, Slamon DJ: Localization of human BRCA1 and its loss in high-grade, non-inherited breast carcinomas. Nat Genet. 1999, 21: 236-240. 10.1038/6029.CrossRefPubMed
37.
Welcsh PL, Owens KN, King MC: Insights into the functions of BRCA1 and BRCA2. Trends Genet. 2000, 16: 69-74. 10.1016/S0168-9525(99)01930-7.CrossRefPubMed
38.
Gowen LC, Avrutskaya AV, Latour AM, Koller BH, Leadon SA: BRCA1 required for transcriptioncoupled repair of oxidative DNA damage. Science. 1998, 281: 1009-1012. 10.1126/science.281.5379.1009.CrossRefPubMed
39.
XL X, Wagner KU, Larson D, Weaver Z, Li C, Ried T, Hennighausen L, Wynshaw-Boris A, Deng CX: Conditional mutation of Brca1 in mammary epithelial cells results in blunted ductal morphogenesis and tumour formation. Nat Genet. 1999, 22: 37-43. 10.1038/8743.CrossRef
40.
Pattingre S, Tassa A, Qu X, Garuti R, Liang XH, Mizushima N, Packer M, Schneider MD, Levine B: Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell. 2005, 122: 927-39. 10.1016/j.cell.2005.07.002.CrossRefPubMed
41.
Liang C, Feng P, Ku B, Dotan I, Canaani D, Oh BH, Jung JU: Autophagic and tumor suppressor activity of a novel Beclin 1-binding protein UVRAG. Nat Cell Biol. 2006, 8: 688-698. 10.1038/ncb1426.CrossRefPubMed
42.
Herman JG, Baylin SB: Gene silencing in cancer in association with promoter hypermethylation. N Engl J Med. 2003, 349: 2042-2054. 10.1056/NEJMra023075.CrossRefPubMed
43.
Douglas DB, Akiyama Y, Carraway H, Belinsky SA, Esteller M, Gabrielson E, Weitzman S, Williams T, Herman JG, Baylin SB: Hypermethylation of a small CpGuanine-rich region correlates with loss of activator protein-2alpha expression during progression of breast cancer. Cancer Res. 2004, 64: 1611-1620. 10.1158/0008-5472.CAN-0318-2.CrossRefPubMed
44.
Unoki M, Nakamura Y: Methylation at CpG islands in intron 1 of EGR2 confers enhancer-like activity. FEBS Lett. 2003, 554: 67-72. 10.1016/S0014-5793(03)01092-5.CrossRefPubMed
45.
Cheng CK, Chow WC, Loo TY, Chan TK, Chan V: The cell cycle checkpoint gene Rad9 is a novel Oncogene activated by 11q13 amplification and DNA methylation in breast cancer. Cancer Res. 2005, 65: 8646-8654. 10.1158/0008-5472.CAN-04-4243.CrossRefPubMed
Metadata
Title
Genetic and epigenetic silencing of the beclin 1gene in sporadic breast tumors
Authors
Zidong Li
Bo Chen
Yiqing Wu
Feng Jin
Yongjing Xia
Xiangjun Liu
Publication date
01-12-2010
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2010
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-10-98

Other articles of this Issue 1/2010

BMC Cancer 1/2010 Go to the issue

Research article

Effect of troglitazone on tumor growth and pulmonary metastasis development of the mouse osteosarcoma cell line LM8

Research article

p21Waf1 expression is regulated by nuclear intermediate filament vimentin in neuroblastoma

Research article

The PI3K/Akt pathway upregulates Id1 and integrin α4 to enhance recruitment of human ovarian cancer endothelial progenitor cells

Research article

Significance of Aurora B overexpression in hepatocellular carcinoma. Aurora B Overexpression in HCC

Research article

The microenvironment determines the breast cancer cells' phenotype: organization of MCF7 cells in 3D cultures

Research article

Cancer resistance of SR/CR mice in the genetic knockout backgrounds of leukocyte effector mechanisms: determinations for functional requirements
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits