Tumor Biology

, Volume 36, Issue 2, pp 1143–1154 | Cite as

Inactivation of PTCH1 is associated with the development of cervical carcinoma: clinical and prognostic implication

  • Chandraditya Chakraborty
  • Sankhadeep Dutta
  • Nupur Mukherjee
  • Sudip Samadder
  • Anirban Roychowdhury
  • Anup Roy
  • Ranajit Kumar Mondal
  • Partha Basu
  • Susanta Roychoudhury
  • Chinmay Kumar Panda
Research Article


The aim of this study was to analyze the alterations of PTCH1 (deletion/promoter methylation/mutation/expression) during the development of cervical cancer (CACX). For this purpose, deletion/methylation of PTCH1 were analyzed in HPV16 positive exfoliated asymptomatic cervical swabs (n = 74), cervical intraepithelial neoplasia (CIN) (n = 32), CACX (n = 174) samples, and two CACX cell lines. The deletion of PTCH1 increased significantly from CIN (11.5 %) to stage I/II (42 %) and comparable in stage III/IV (46 %). Low frequency (14–16 %) of PTCH1 methylation was seen in the asymptomatic exfoliated cervical cells and in the normal epithelium adjacent to the tumor followed by a significant increase in CIN (31 %) to stage I/II (57 %) and comparable in stage III/IV (58 %). The overall alterations (deletion/methylation) of PTCH1 significantly increased from CIN (34 %) to stage I/II (70 %) and comparable in stage III/IV (69 %). Interestingly, in the normal epithelium, methylation of PTCH1 was high in basal/parabasal layers (83 %), followed by decrease in the spinous layer (33 %), and showed significant inverse correlation with its expression. Reduced expression of PTCH1 seen in tumors showed a significant association with its alterations (deletion/methylation). The expression pattern of PTCH1 showed an inverse correlation with the nuclear expression of GLI1 in the normal epithelium as well as in the tumors. High nuclear expression of HPV16, E6, and E7 were seen in basal/parabasal layers of the normal epithelium and also in tumors. The PTCH1 alterations (deletion and/or methylation) in tumors and its methylation in adjacent normal epithelium were associated with poor prognosis of patients. Thus, our data suggests that activation of the Hedgehog pathway due to PTCH1 inactivation along with HPV infection is important in CACX development.


PTCH1 methylation Adjacent normal Expression CACX progression HPV 



The authors thank the Director, Chittaranjan National Cancer Institute, Kolkata, India. They also thank Dr. Shyamsundar Mandal, Chittaranjan National Cancer Institute, for his assistance in solving statistical problems. They are also grateful to Professor (Dr.) H. zur Hausen and Professor (Dr. Mrs.) E.M. de Villiers for their generous gift of HPV16/18 plasmids. This work was supported by CSIR-JRF/NET grant [File No.09/030(0059)/2010-EMR-I] to Mr C. Chakraborty, CSIR-SRF grant [File No. 09/030(0065)/2011-EMR-I] to Mr S. Dutta, and CSIR-RA grant [File No.09/030(0071)/2013-EMRI] to Dr. N. Mukherjee from Council of Scientific and Industrial Research (CSIR), UGC-JRF/NET grant [Sr No. 2121130723] to Mr S. Samadder from University Grants Commission (UGC), Government of India, and grant [SR/SO/HS-116/2007] from the Department of Science and Technology (DST), Government of India, to Dr. C. K. Panda.

Conflicts of interest


Supplementary material

13277_2014_2707_MOESM1_ESM.pdf (1.6 mb)
ESM 1 (PDF 1631 kb)


  1. 1.
    Arbyn M. Worldwide burden of cervical cancer in 2008. Ann Oncol. 2011;22(12):2675–86.CrossRefPubMedGoogle Scholar
  2. 2.
    WHO/ICO Information Centre on Human Papilloma Virus (HPV) and Related Diseases report in India. Summary Report 2014Google Scholar
  3. 3.
    Munoz N, Bosch FX, de Sanjose S, Herrero R, Castellsague X, Shah KV, et al. Epidemiologic classification of human papillomavirus types associated with cervical cancer. N Engl J Med. 2003;348:518–27.CrossRefPubMedGoogle Scholar
  4. 4.
    Dutta S, Begum R, Mazumder Indra D, Mandal SS, Mondal R, Biswas J, et al. Prevalence of human papillomavirus in women without cervical cancer: a population-based study in Eastern India. Int J Gynecol Pathol. 2012;31:178–83.CrossRefPubMedGoogle Scholar
  5. 5.
    WHO/ICO Information Centre on Human Papilloma Virus (HPV) and Related Diseases report in World. Summary Report 2014Google Scholar
  6. 6.
    Bosch FX, de Sanjose S. Chapter 1: human papillomavirus and cervical cancer–burden and assessment of causality. J Natl Cancer Inst. 2003;31:3–13.CrossRefGoogle Scholar
  7. 7.
    Snijders PJ, Steenbergen RD, Heideman DA, Meijer CJ. HPV-mediated cervical carcinogenesis: concepts and clinical implications. J Pathol. 2006;208:152–64.CrossRefPubMedGoogle Scholar
  8. 8.
    Lopez J, Ruiz G, Organista-Nava J, Gariglio P, Garcia-Carranca A. Human papillomavirus infections and cancer stem cells of tumors from the uterine cervix. Open Virol J. 2012;6:232–40.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Talora C, Sgroi DC, Crum CP, Dotto GP. Specific down-modulation of Notch1 signaling in cervical cancer cells is required for sustained HPV-E6/E7 expression and late steps of malignant transformation. Genes Dev. 2002;16:2252–63.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Uren A, Fallen S, Yuan H, Usubutun A, Kucukali T, Schlegel R, et al. Activation of the canonical Wnt pathway during genital keratinocyte transformation: a model for cervical cancer progression. Cancer Res. 2005;65:6199–206.CrossRefPubMedGoogle Scholar
  11. 11.
    Chaudary N, Pintilie M, Hedley D, Fyles AW, Milosevic M, Clarke B, et al. Hedgehog pathway signaling in cervical carcinoma and outcome after chemoradiation. Cancer. 2012;118:3105–15.CrossRefPubMedGoogle Scholar
  12. 12.
    Xuan YH, Jung HS, Choi YL, Shin YK, Kim HJ, Kim KH, et al. Enhanced expression of hedgehog signaling molecules in squamous cell carcinoma of uterine cervix and its precursor lesions. Mod Pathol. 2006;19:1139–47.PubMedGoogle Scholar
  13. 13.
    Agren M, Kogerman P, Kleman MI, Wessling M, Toftgard R. Expression of the PTCH1 tumor suppressor gene is regulated by alternative promoters and a single functional Gli-binding site. Gene. 2004;330:101–14.CrossRefPubMedGoogle Scholar
  14. 14.
    Daya-Grosjean L, Couve-Privat S. Sonic hedgehog signaling in basal cell carcinoma. Cancer Lett. 2005;225:181–92.CrossRefPubMedGoogle Scholar
  15. 15.
    Ghosh A, Ghosh S, Maiti GP, Mukherjee S, Mukherjee N, Chakraborty J, et al. Association of FANCC and PTCH1 with the development of early dysplastic lesions of the head and neck. Ann Surg Oncol. 2010;19:S528–38.CrossRefGoogle Scholar
  16. 16.
    Wang XD, Inzunza H, Chang H, Qi Z, Hu B, Malone D, et al. Mutations in the hedgehog pathway genes SMO and PTCH1 in human gastric tumors. PLoS One. 2013;8:e54415.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Lu X, Nikaido T, Toki T, Zhai YL, Kita N, Konishi I, et al. Loss of heterozygosity among tumor suppressor genes in invasive and in situ carcinoma of the uterine cervix. Int J Gynecol Cancer. 2000;10:452–8.CrossRefPubMedGoogle Scholar
  18. 18.
    Lof-Ohlin ZM, Levanat S, Sabol M, Sorbe B, Nilsson TK. Promoter methylation in the PTCH gene in cervical epithelial cancer and ovarian cancer tissue as studied by eight novel Pyrosequencing(R) assays. Int J Oncol. 2011;38:685–92.PubMedGoogle Scholar
  19. 19.
    Sinha S, Singh RK, Alam N, Roy A, Roychoudhury S, Panda CK. Alterations in candidate genes PHF2, FANCC, PTCH1 and XPA at chromosomal 9q22.3 region: pathological significance in early- and late-onset breast carcinoma. Mol Cancer. 2008;7:84.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Dasgupta S, Mukherjee N, Roy S, Roy A, Sengupta A, Roychowdhury S, et al. Mapping of the candidate tumor suppressor genes’ loci on human chromosome 3 in head and neck squamous cell carcinoma of an Indian patient population. Oral Oncol. 2002;38:6–15.CrossRefPubMedGoogle Scholar
  21. 21.
    Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual. 2nd ed. NY: Cold Spring Harbor Press; 1989.Google Scholar
  22. 22.
    Singh RK, Maulik S, Mitra S, Mondal RK, Basu PS, Roychowdhury S, et al. Human papillomavirus prevalence in postradiotherapy uterine cervical carcinoma patients: correlation with recurrence of the disease. Int J Gynecol Cancer. 2006;16:1048–54.CrossRefPubMedGoogle Scholar
  23. 23.
    Loginov VI, Maliukova AV, Seregin Iu A, Khodyrev DS, Kazubskaia TP, Ermilova VD, et al. Methylation of the promoter region of the RASSF1A gene, a candidate tumor suppressor, in primary epithelial tumors. Mol Biol. 2004;38:654–67.CrossRefGoogle Scholar
  24. 24.
    Ivanova T, Petrenko A, Gritsko T, Vinokourova S, Eshilev E, Kobzeva V, et al. Methylation and silencing of the retinoic acid receptor-b2 gene in cervical cancer. BMC Cancer. 2002;2:4.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Tripathi A, Banerjee S, Chunder N, et al. Differential alterations of the genes in the CDKN2A-CCND1-CDK4-RB1 pathway are associated with the development of head and neck squamous cell carcinoma in Indian patients. J Cancer Res Clin Oncol. 2003;129:642–50.CrossRefGoogle Scholar
  26. 26.
    Ghosh S, Ghosh A, Maiti GP, Alam N, Roy A, Roy B, et al. Alterations of 3p21.31 tumor suppressor genes in head and neck squamous cell carcinoma: correlation with progression and prognosis. Int J Cancer. 2008;123:2594–604.CrossRefPubMedGoogle Scholar
  27. 27.
    Perrone F, Suardi S, Pastore E, Casieri P, Orsenigo M, Caramuta S, et al. Molecular and cytogenetic subgroups of oropharyngeal squamous cell carcinoma. Clin Cancer Res. 2006;12:6643–51.CrossRefPubMedGoogle Scholar
  28. 28.
    Samarzija I, Beard P. Hedgehog pathway regulators influence cervical cancer cell proliferation, survival and migration. Biochem Biophys Res Commun. 2012;425:64–9.CrossRefPubMedGoogle Scholar
  29. 29.
    Ressler S, Scheiden R, Dreier K, Laich A, Müller-Holzner E, Pircher H, et al. High-risk human papillomavirus E7 oncoprotein detection in cervical squamous cell carcinoma. Clin Cancer Res. 2007;23:7067–72.CrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Chandraditya Chakraborty
    • 1
  • Sankhadeep Dutta
    • 1
  • Nupur Mukherjee
    • 1
  • Sudip Samadder
    • 1
  • Anirban Roychowdhury
    • 1
  • Anup Roy
    • 2
  • Ranajit Kumar Mondal
    • 3
  • Partha Basu
    • 3
  • Susanta Roychoudhury
    • 4
  • Chinmay Kumar Panda
    • 1
  1. 1.Department of Oncogene RegulationChittaranjan National Cancer InstituteKolkataIndia
  2. 2.North Bengal Medical College and HospitalSiliguriIndia
  3. 3.Department of Gynecological OncologyChittaranjan National Cancer InstituteKolkataIndia
  4. 4.Cancer Biology & Inflammatory Disorder DivisionIndian Institute of Chemical BiologyKolkataIndia

Personalised recommendations