Tumor Biology

, Volume 36, Issue 3, pp 1529–1537 | Cite as

ATM may be a protective factor in endometrial carcinogenesis with the progesterone pathway

  • Weiwei Shan
  • Chao Wang
  • Zhenbo Zhang
  • Xuezhen Luo
  • Chengcheng Ning
  • Yinhua Yu
  • Youji Feng
  • Chao Gu
  • Xiaojun Chen
Research Article


The purpose of the study was to explore the role and mechanism of ataxia-telangiectasia mutated (ATM) protein in endometrial carcinogenesis. A reverse-phase protein array (RPPA) was used to analyze the expression of ATM signal pathway proteins in Ishikawa and progesterone-insensitive Ishikawa. ATM expression was detected in endometrium specimens by immunohistochemistry, including 8 cases with proliferative endometrium, 6 cases with secretory endometrium, 10 cases with simple hyperplasia (SH), 13 cases of complex hyperplasia (CH), 11 cases of endometrial atypical hyperplasia (EAH), and 83 cases with type I endometrial cancer. The relationship between ATM expression and other clinicopathological indicators was also examined in type I endometrial cancer patients. The mechanisms of ATM were explored in vitro with the endometrial cell lines Ishikawa and RL95-2. A cell counting kit-8 (CCK-8) test and Western blot analysis were performed to test proliferation and protein expression. Statistical analysis was performed with SPSS19.0. The significance level was set at 0.05. ATM was increased with medroxyprogesterone acetate (MPA) stimulation in Ishikawa in RPPA. ATM expression gradually decreased in endometrial hyperplasic lesions compared with the normal proliferative and secretory endometrium and was the lowest in type I endometrial cancer. ATM expression was negatively correlated with pathological grades in type I endometrial cancer. In vitro, ATM silencing retarded proliferation inhibition in Ishikawa and RL95-2 treated with MPA. ATM silencing could down-regulate the MPA-stimulated signal proteins, including Chk2, P53, and caspase-3 in vitro. MPA might exert its role through activating the ATM-associated pathway, ATM-Chk2-P53-caspase-3 (active), preserving normal endometrium and protecting it from malignancies. ATM might be a promising indicator for endometrial hyperplasia and cancer.


ATM Endometrial cancer Progesterone 



This study was supported by the Shanghai Municipal Science Foundation 2011 (Project No 11ZR1404300); the National Natural Science Foundation of China, 2012 (NSFC No 81101953); and the Shanghai Municipal Science Foundation 2013 (Project No 134119a4500).

Conflicts of interest


Supplementary material

13277_2014_2712_MOESM1_ESM.xlsx (14 kb)
S1 A total of 183 antibodies were applied in the RPPA analysis. (XLSX 14 kb)
13277_2014_2712_Fig7_ESM.gif (65 kb)

Cell apoptosis in Ishikawa by flow cytometry. Compared to the NC group, NC+MPA and SI+MPA groups had higher apoptosis rates. The comparison was performed with Student’s t-test. NC, negative control. SI, ATM-siRNA, * P < 0.05, **P < 0.01. (GIF 65 kb)

13277_2014_2712_MOESM2_ESM.tif (1 mb)
High resolution image (TIFF 1037 kb)
13277_2014_2712_Fig8_ESM.gif (57 kb)

Cell apoptosis in Ishikawa cells by flow cytometry. Compared to the NC group, the SI, NC+MPA and SI+MPA groups had higher apoptosis rates. Student’s t-test was used for the evaluation. NC, negative control. SI, ATM-siRNA, * P < 0.05,**P < 0.01. (GIF 57 kb)

13277_2014_2712_MOESM3_ESM.tif (1 mb)
High resolution image (TIFF 1030 kb)


  1. 1.
    Weiderpass E, Labreche F. Malignant tumors of the female reproductive system. Saf Health Work. 2012;3:166–80.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Pfeiffer RM, Park Y, Kreimer AR, Lacey JJ, Pee D, Greenlee RT, et al. Risk prediction for breast, endometrial, and ovarian cancer in white women aged 50 y or older: derivation and validation from population-based cohort studies. PLoS Med. 2013;10:e1001492.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Le Gallo M, Bell DW. The emerging genomic landscape of endometrial cancer. Clin Chem. 2014;60:98–110.CrossRefPubMedGoogle Scholar
  4. 4.
    Li X, Shao R. PCOS and obesity: insulin resistance might be a common etiology for the development of type I endometrial carcinoma. Am J Cancer Res. 2014;4:73–9.PubMedPubMedCentralGoogle Scholar
  5. 5.
    Kim JJ, Kurita T, Bulun SE. Progesterone action in endometrial cancer, endometriosis, uterine fibroids, and breast cancer. Endocr Rev. 2013;34:130–62.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Smith J, Tho LM, Xu N, Gillespie DA. The ATM-Chk2 and ATR-Chk1 pathways in DNA damage signaling and cancer. Adv Cancer Res. 2010;108:73–112.CrossRefPubMedGoogle Scholar
  7. 7.
    Stracker TH, Roig I, Knobel PA, Marjanovic M. The ATM signaling network in development and disease. Front Genet. 2013;4:37.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Fan C, Quan R, Feng X, Gillis A, He L, Matsumoto ED, et al. ATM activation is accompanied with earlier stages of prostate tumorigenesis. Biochim Biophys Acta. 2006;1763:1090–7.CrossRefPubMedGoogle Scholar
  9. 9.
    Puccini J, Shalini S, Voss AK, Gatei M, Wilson CH, Hiwase DK, et al. Loss of caspase-2 augments lymphomagenesis and enhances genomic instability in Atm-deficient mice. Proc Natl Acad Sci U S A. 2013;110:19920–5.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Selvarajah J, Nathawat K, Moumen A, Ashcroft M, Carroll VA. Chemotherapy-mediated p53-dependent DNA damage response in clear cell renal cell carcinoma: role of the mTORC1/2 and hypoxia-inducible factor pathways. Cell Death Dis. 2013;4:e865.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Takagi M, Sato M, Piao J, Miyamoto S, Isoda T, Kitagawa M, et al. ATM-dependent DNA damage-response pathway as a determinant in chronic myelogenous leukemia. DNA Repair (Amst). 2013;12:500–7.CrossRefGoogle Scholar
  12. 12.
    Vang R, Gown AM, Barry TS, Wheeler DT, Ronnett BM. Immunohistochemistry for estrogen and progesterone receptors in the distinction of primary and metastatic mucinous tumors in the ovary: an analysis of 124 cases. Mod Pathol. 2006;19:97–105.CrossRefPubMedGoogle Scholar
  13. 13.
    Zhao S, Li G, Yang L, Li L, Li H. Response-specific progestin resistance in a newly characterized Ishikawa human endometrial cancer subcell line resulting from long-term exposure to medroxyprogesterone acetate. Oncol Lett. 2013;5:139–44.PubMedGoogle Scholar
  14. 14.
    Zhang L, Wei Q, Mao L, Liu W, Mills GB, Coombes K. Serial dilution curve: a new method for analysis of reverse phase protein array data. Bioinformatics. 2009;25:650–4.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Tsuda H, Hashiguchi Y, Inoue T, Yamamoto K. Alteration of G2 cell cycle regulators occurs during carcinogenesis of the endometrium. Oncol-Basel. 2003;65:159–66.CrossRefGoogle Scholar
  16. 16.
    Einarsdottir K, Humphreys K, Bonnard C, Li Y, Li Y, Chia KS, et al. Effect of ATM, CHEK2 and ERBB2 TAGSNPs and haplotypes on endometrial cancer risk. Hum Mol Genet. 2007;16:154–64.CrossRefPubMedGoogle Scholar
  17. 17.
    Mirzayans R, Andrais B, Scott A, Murray D. New insights into p53 signaling and cancer cell response to DNA damage: implications for cancer therapy. J Biomed Biotechnol. 2012;2012:170325.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Choi JD, Lee JS. Interplay between epigenetics and genetics in cancer. Genomics Inform. 2013;11:164–73.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Masai H. ATM in prevention of genomic instability. Cell Cycle. 2014;13(6):882–3.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Kim M, Park HJ, Seol JW, Jang JY, Cho YS, Kim KR, et al. VEGF-A regulated by progesterone governs uterine angiogenesis and vascular remodelling during pregnancy. EMBO Mol Med. 2013;5:1415–30.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Wang J, Taylor A, Showeil R, Trivedi P, Horimoto Y, Bagwan I, et al. Expression profiling and significance of VEGF-A, VEGFR2, VEGFR3 and related proteins in endometrial carcinoma. Cytokine. 2014;68:94–100.CrossRefPubMedGoogle Scholar
  22. 22.
    Gu C, Zhang Z, Yu Y, Liu Y, Zhao F, Yin L, et al. Inhibiting the PI3K/Akt pathway reversed progestin resistance in endometrial cancer. Cancer Sci. 2011;102:557–64.CrossRefPubMedGoogle Scholar
  23. 23.
    Kauff ND. ATR mutations in endometrial cancer: a window into the role of mismatch repair defects. J Clin Oncol. 2009;27:3077–8.CrossRefPubMedGoogle Scholar
  24. 24.
    Banno K, Kisu I, Yanokura M, Tsuji K, Masuda K, Ueki A, et al. Biomarkers in endometrial cancer: possible clinical applications (review). Oncol Lett. 2012;3:1175–80.PubMedPubMedCentralGoogle Scholar
  25. 25.
    Tommiska J, Bartkova J, Heinonen M, Hautala L, Kilpivaara O, Eerola H, et al. The DNA damage signalling kinase ATM is aberrantly reduced or lost in BRCA1/BRCA2-deficient and ER/PR/ERBB2-triple-negative breast cancer. Oncogene. 2008;27:2501–6.CrossRefPubMedGoogle Scholar
  26. 26.
    Liang Y, Lin SY, Brunicardi FC, Goss J, Li K. DNA damage response pathways in tumor suppression and cancer treatment. World J Surg. 2009;33:661–6.CrossRefPubMedGoogle Scholar
  27. 27.
    Yang DQ, Halaby MJ, Li Y, Hibma JC, Burn P. Cytoplasmic ATM protein kinase: an emerging therapeutic target for diabetes, cancer and neuronal degeneration. Drug Discov Today. 2011;16:332–8.CrossRefPubMedGoogle Scholar
  28. 28.
    Kim JW, Im SA, Kim MA, Cho HJ, Lee DW, Lee KH, et al. Ataxia-telangiectasia-mutated protein expression with microsatellite instability in gastric cancer as prognostic marker. Int J Cancer. 2014;134:72–80.CrossRefPubMedGoogle Scholar
  29. 29.
    Guo X, Yang C, Qian X, Lei T, Li Y, Shen H, et al. Estrogen receptor alpha regulates ATM expression through miRNAs in breast cancer. Clin Cancer Res. 2013;19:4994–5002.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Wilk A, Waligorska A, Waligorski P, Ochoa A, Reiss K. Inhibition of ERbeta induces resistance to cisplatin by enhancing Rad51-mediated DNA repair in human medulloblastoma cell lines. PLoS One. 2012;7:e33867.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Bozhanov SS, Angelova SG, Krasteva ME, Markov TL, Christova SL, Gavrilov IG, et al. Alterations in p53, BRCA1, ATM, PIK3CA, and HER2 genes and their effect in modifying clinicopathological characteristics and overall survival of Bulgarian patients with breast cancer. J Cancer Res Clin Oncol. 2010;136:1657–69.CrossRefPubMedGoogle Scholar
  32. 32.
    Mhawech-Fauceglia P, Wang D, Kim G, Sharifian M, Chen X, Liu Q, et al. Expression of DNA repair proteins in endometrial cancer predicts disease outcome. Gynecol Oncol. 2014;132:593–8.CrossRefPubMedGoogle Scholar
  33. 33.
    Fang S, Krahe R, Lozano G, Han Y, Chen W, Post SM, et al. Effects of MDM2, MDM4 and TP53 codon 72 polymorphisms on cancer risk in a cohort study of carriers of TP53 germline mutations. PLoS One. 2010;5:e10813.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Hirshfield KM, Rebbeck TR, Levine AJ. Germline mutations and polymorphisms in the origins of cancers in women. J Oncol. 2010;2010:297671.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Lei W, Feng XH, Deng WB, Ni H, Zhang ZR, Jia B, et al. Progesterone and DNA damage encourage uterine cell proliferation and decidualization through up-regulating ribonucleotide reductase 2 expression during early pregnancy in mice. J Biol Chem. 2012;287:15174–92.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Sun Y, Jiang X, Price BD. Tip60: connecting chromatin to DNA damage signaling. Cell Cycle. 2010;9:930–6.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Xu Y, Tong J, Ai Z, Wang J, Teng Y. Epidermal growth factor receptor signaling pathway involved in progestin-resistance of human endometrial carcinoma: in a mouse model. J Obstet Gynaecol Res. 2012;38:1358–66.CrossRefPubMedGoogle Scholar
  38. 38.
    Arnold JT, Lessey BA, Seppala M, Kaufman DG. Effect of normal endometrial stroma on growth and differentiation in Ishikawa endometrial adenocarcinoma cells. Cancer Res. 2002;62:79–88.PubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Weiwei Shan
    • 1
    • 2
  • Chao Wang
    • 3
  • Zhenbo Zhang
    • 4
  • Xuezhen Luo
    • 1
    • 2
  • Chengcheng Ning
    • 1
    • 2
  • Yinhua Yu
    • 1
    • 2
  • Youji Feng
    • 4
  • Chao Gu
    • 1
    • 2
    • 5
  • Xiaojun Chen
    • 1
    • 2
    • 5
  1. 1.Department of GynecologyObstetrics and Gynecology Hospital of Fudan UniversityShanghaiPeople’s Republic of China
  2. 2.Shanghai Key Laboratory of Female Reproductive Endocrine Related DiseasesShanghaiChina
  3. 3.Department of PathologyObstetrics and Gynecology Hospital of Fudan UniversityShanghaiPeople’s Republic of China
  4. 4.Department of GynecologyShanghai First People’s Hospital Affiliated to Shanghai Jiao tong UniversityShanghaiChina
  5. 5.ShanghaiPeople’s Republic of China

Personalised recommendations