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High nuclear expression of DNMT1 in correlation with inactivation of TET1 portray worst prognosis among the cervical carcinoma patients: clinical implications

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Abstract

In this study, we aimed to understand the interplay of the epigenetic modifier genes DNMT1 and TET1 along with HPV infection in the cervical epithelium and how it changes during tumorigenesis. For this purpose, initially the bioinformatical analysis (methylation and expression profile) of DNMT1 and TET1 was analyzed in the TCGA dataset. Next genetic (deletion) and epigenetic profiling (promoter methylation) of DNMT1 and TET1 were done in our sample pool and also validated in CACX cell lines as well. The results were further correlated with different clinicopathological parameters. Our data revealed that HPV infection in basal/parabasal layers of cervical epithelium actually disrupts the epigenetic homeostasis of DNMT1 and TET1 proteins which ultimately leads to the high expression of DNMT1 along with further reduction in TET1 protein during the development of carcinoma. Further, in-depth look into the results revealed that comparatively low methylation frequency of DNMT1 coupled with high promoter methylation and deletion frequency [22–46%] of TET1 were the plausible reasons of their antagonistic expression profile during the progression of the disease. Interestingly, the prevalence of DNMT1 [9.1%] and TET1 promoter methylation [22.7%] found in both the plasma DNA of the respective CACX patients implicated its diagnostic importance in this study. Lastly, molecular alteration of TET1 alone or in combination with DNMT1 showed the worst overall survival among the patients. Hence, it may be concluded that an inverse molecular profile of DNMT1 and TET1 genes seen in the proliferative basal-parabasal layers of the cervical epithelium was aggravated during the development of CACX along with genetic and epigenetic changes due to HPV infection.

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Abbreviations

HPV:

Human papilloma virus

CIN:

Cervical intraepithelial neoplasia

CACX:

Cervical carcinoma

DNMT1:

DNA methyl transferase 1

TET1:

Ten eleven translocase 1

mRNA:

Messenger RNA

qRT-PCR:

Quantitative real time PCR

5-aza-Dc:

5-Aza-2’-deoxycytidine

References

  • Arbyn M, Castellsague X, de Sanjose S, Bruni L, Saraiya M, Bray F et al (2011) Worldwide burden of cervical cancer in 2008. Ann Oncol 22:2675–2686. https://doi.org/10.1093/annonc/mdr015

    Article  CAS  Google Scholar 

  • Au Yeung CL, Tsang WP, Tsang TY, Co NN, Yau PL, Kwok TT (2010) HPV-16 E6 upregulation of DNMT1 through repression of tumour suppressor p53. Oncol Rep 24(6):1599–1604. https://doi.org/10.3892/or_00001023

    Article  CAS  Google Scholar 

  • Basu M, Chakroborty B, Ghosh S, Samadder S, Dutta S, Roy A, Pal DK, Ghosh A, Panda CK (2020) Divergent molecular profle of PIK3CA gene in arsenic associated bladder carcinoma: clinical importance. Mutagenesis 35(6):499–508. https://doi.org/10.1093/mutage/geaa031

    Article  CAS  Google Scholar 

  • Basu M, Ghosh S, Roychowdhury A, Samadder S, Das P, Addya S, Roy A, Pal DK, Roychoudhury S, Ghosh A, Panda CK (2020) Integrative genomics and pathway analysis identified prevalent FA-BRCA pathway alterations in arsenic-associated urinary bladder carcinoma: Chronic arsenic accumulation in cancer tissues hampers the FA-BRCA pathway. Genomics 112(6):5055–5065. https://doi.org/10.1016/j.ygeno.2020.09.012

    Article  CAS  Google Scholar 

  • Burley M, Roberts S, Parish JL (2020) Epigenetic regulation of human papillomavirus transcription in the productive virus life cycle. Semin Immunopathol 42:159–171. https://doi.org/10.1007/s00281-019-00773-0

    Article  CAS  Google Scholar 

  • Cao J, Dong J, Wang Y, Chen Y (2019) The expressions of DNA methyltransferase 1 (DNMT1) and cyclin A1 (CCNA1) in cervical carcinogenesis. Int J Clin Exp Pathol 12(1):40–49

    CAS  Google Scholar 

  • Chakraborty C, Dutta S, Mukherjee N, Samadder S, Roychowdhury A, Roy A, Mondal RK, Basu P, Roychoudhury S, Panda CK (2015) Inactivation of PTCH1 is associated with the development of cervical carcinoma: clinical and prognostic implication. Tumour Biol 36(2):1143–1154. https://doi.org/10.1007/s13277-014-2707-1

    Article  CAS  Google Scholar 

  • Chakraborty C, Samadder S, Roychowdhury A, Roy A, Das P, Mandal RK, Roychoudhury S, Panda CK (2018) Activation of Wnt-β-catenin pathway in basal-parabasal layers of normal cervical epithelium comparable during development of uterine cervical carcinoma. Mol Cell Biochem 443(1–2):121–130. https://doi.org/10.1007/s11010-017-3216-5

    Article  CAS  Google Scholar 

  • Dasgupta H, Islam S, Alam N, Roy A, Roychoudhury S, Panda CK (2019a) Hypomethylation of mismatch repair genes MLH1 and MSH2 is associated with chemotolerance of breast carcinoma: Clinical significance. J Surg Oncol 119(1):88–100. https://doi.org/10.1002/jso.25304

    Article  CAS  Google Scholar 

  • Dasgupta H, Islam MS, Alam N, Roy A, Roychoudhury S, Panda CK (2019b) Induction of HRR genes and inhibition of DNMT1 is associated with anthracycline anti-tumor antibiotic-tolerant breast carcinoma cells. Mol Cell Biochem 453(1–2):163–178. https://doi.org/10.1007/s11010-018-3442-5

    Article  CAS  Google Scholar 

  • Dasgupta S, Mukherjee N, Roy S, Roy A, Sengupta A, Roychowdhury S et al (2002) Mapping of the candidate tumour suppressor genes’ loci on human chromosome 3 in head and neck squamous cell carcinoma of an Indian patient population. Oral Oncol 38:6–15. https://doi.org/10.1016/S1368-8375(00)00131-7

    Article  CAS  Google Scholar 

  • Dueñas-González, A., Lizano, M., Candelaria, M. et al. (2005) Epigenetics of cervical cancer. An overview and therapeutic perspectives. Mol Cancer 4, 38. https://doi.org/10.1186/1476-4598-4-38.

  • Feng C, Dong J, Chang W, Cui M, Xu T (2018) The progress of methylation regulation in gene expression of cervical cancer. Int J Genomics 2018:8260652. https://doi.org/10.1155/2018/8260652

    Article  CAS  Google Scholar 

  • Ghosh S, Ghosh A, Maiti GP, Alam N, Roy A, Roy B et al (2008) Alterations of 3p21.31 tumour suppressor genes in head and neck squamous cell carcinoma: correlation with progression and prognosis. Int J Cancer 123:2594–2604. https://doi.org/10.1002/ijc.23834

    Article  CAS  Google Scholar 

  • IARC/ICO Information Centre on Human Papilloma Virus (HPV) and Related Diseases report in India. Summary Report. 2021

  • Islam S, Dasgupta H, Basu M, Roy A, Alam N, Roychoudhury S, Panda CK (2020) Downregulation of beta-catenin in chemo-tolerant TNBC through changes in receptor and antagonist profiles of the WNT pathway: clinical and prognostic implications. Cell Oncol (dordr) 43(4):725–741. https://doi.org/10.1007/s13402-020-00525-5

    Article  CAS  Google Scholar 

  • Li Z, Li Y, Li Y, Ren K, Li X, Han X, Wang J (2017) Long non-coding RNA H19 promotes the proliferation and invasion of breast cancer through upregulating DNMT1 expression by sponging miR-152. J Biochem Mol Toxicol. https://doi.org/10.1002/jbt.21933

    Article  Google Scholar 

  • López J, Ruíz G, Organista-Nava J, Gariglio P, García-Carrancá A (2012) Human papillomavirus infections and cancer stem cells of tumours from the uterine cervix. Open Virol J 6:232–240. https://doi.org/10.2174/1874357901206010232

    Article  Google Scholar 

  • Łuczak MW, Roszak A, Pawlik P, Kędzia H, Kędzia W, Malkowska-Walczak B, Jagodziński PP (2012) Transcriptional analysis of CXCR4, DNMT3A, DNMT3B and DNMT1 gene expression in primary advanced uterine cervical carcinoma. Int J Oncol 40:860–866. https://doi.org/10.3892/ijo.2011.1183

    Article  CAS  Google Scholar 

  • Malisic E, Brotto K, Krivokuca A, Cavic M, Jankovic R (2014) Overall human papilloma virus and types 16/18 prevalence in women with normal cervical cytology in Serbia: is it time for human papillomavirus testing and/or vaccination? J BUON. 19(4):973–9

    Google Scholar 

  • Nuryadi E, Sasaki Y, Hagiwara Y, Permata T, Sato H, Komatsu S, Yoshimoto Y, Murata K, Ando K, Kubo N, Okonogi N, Takakusagi Y, Adachi A, Iwanaga M, Tsuchida K, Tamaki T, Noda SE, Hirota Y, Shibata A, Ohno T, Nakano T (2018) Mutational analysis of uterine cervical cancer that survived multiple rounds of radiotherapy. Oncotarget 9(66):32642–32652. https://doi.org/10.18632/oncotarget.25982

    Article  Google Scholar 

  • Paschos K, Allday MJ (2010) Epigenetic reprogramming of host genes in viral and microbial pathogenesis. Trends Microbiol 18(10):439–447. https://doi.org/10.1016/j.tim.2010.07.003

    Article  CAS  Google Scholar 

  • Perrone F, Suardi S, Pastore E, Casieri P, Orsenigo M, Caramuta S et al (2006) Molecular and cytogenetic subgroups of oropharyngeal squamous cell carcinoma. Clin Cancer Res 12:6643–6651. https://doi.org/10.1158/1078-0432.CCR-06-1759

    Article  CAS  Google Scholar 

  • Piyathilake CJ, Badiga S, Borak SG, Weragoda J, Bae S, Matthews R, Bell WC, Partridge EE (2017) A higher degree of expression of DNA methyl transferase 1 in cervical cancer is associated with poor survival outcome. Int J Women’s Health 9:413–420. https://doi.org/10.2147/IJWH.S133441

    Article  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor, Cold Spring Harbor Laboratory

    Google Scholar 

  • Sambrook J, Russell DW, Sambrook J (2006) The condensed protocols from molecular cloning: a laboratory manual, vol 800. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Sawada M, Kanai Y, Arai E, Ushijima S, Ojima H, Hirohashi S (2007) Increased expression of DNA methyltransferase 1 (DNMT1) protein in uterine cervix squamous cell carcinoma and its precursor lesion. Cancer Lett 251(2):211–219. https://doi.org/10.1016/j.canlet.2006.11.023

    Article  CAS  Google Scholar 

  • Shi JF, Li XJ, Si XX, Li AD, Ding HJ, Han X, Sun YJ (2012) ERα positively regulated DNMT1 expression by binding to the gene promoter region in human breast cancer MCF-7 cells. Biochem Biophys Res Commun 427(1):47–53. https://doi.org/10.1016/j.bbrc.2012.08.144

    Article  CAS  Google Scholar 

  • Singh RK, Maulik S, Mitra S, Mondal RK, Basu PS, Roychowdhury S et al (2006) Human papillomavirus prevalence in postradiotherapy uterine cervical carcinoma patients: correlation with recurrence of the disease. Int J Gynecol Cancer 16:1048–1054. https://doi.org/10.1111/j.1525-1438.2006.00550

    Article  CAS  Google Scholar 

  • Snijders PJF, Steenbergen RDM, Heideman DAM, Meijer CJLM (2006) HPV-mediated cervical carcinogenesis: concepts and clinical implications. J Pathol 208:152–164. https://doi.org/10.1002/path.1866

    Article  CAS  Google Scholar 

  • Stiasny A, Kuhn C, Mayr D, Alexiou C, Janko C, Wiest I, Jeschke U, Kost B (2016) Immunohistochemical evaluation of E6/E7 HPV oncoproteins staining in cervical cancer. Anticancer Res 36(6):3195–3198

    CAS  Google Scholar 

  • Su PH, Hsu YW, Huang RL, Chen LY, Chao TK, Liao CC, Chen CW, Wu TI, Mao SP, Balch C, Lai HC (2019) TET1 promotes 5hmC-dependent stemness, and inhibits a 5hmC-independent epithelial-mesenchymal transition, in cervical precancerous lesions. Cancer Lett 450:53–62. https://doi.org/10.1016/j.canlet.2019.01.033

    Article  CAS  Google Scholar 

  • Thomson JP, Ottaviano R, Unterberger EB, Lempiäinen H, Muller A, Terranova R, Illingworth RS, Webb S, Kerr AR, Lyall MJ, Drake AJ, Wolf CR, Moggs JG, Schwarz M, Meehan RR (2016) Loss of Tet1-associated 5-hydroxymethylcytosine is concomitant with aberrant promoter hypermethylation in liver cancer. Can Res 76(10):3097–3108. https://doi.org/10.1158/0008-5472.CAN-15-1910

    Article  CAS  Google Scholar 

  • Wang X, Li B (2017) DNMT1 regulates human endometrial carcinoma cell proliferation. Onco Targets Ther 10:1865–1873. https://doi.org/10.2147/OTT.S130022

    Article  CAS  Google Scholar 

  • Yang HJ, Liu VW, Wang Y, Chan KY, Tsang PC, Khoo US, Cheung AN, Ngan HY (2004) Detection of hypermethylated genes in tumor and plasma of cervical cancer patients. Gynecol Oncol 93(2):435–440. https://doi.org/10.1016/j.ygyno.2004.01.039

    Article  CAS  Google Scholar 

  • Yim EK, Park JS (2005) The role of HPV E6 and E7 oncoproteins in HPV-associated cervical carcinogenesis. Cancer Res Treat 37(6):319–324. https://doi.org/10.4143/crt.2005.37.6.319

    Article  Google Scholar 

  • Yu Y, Qi J, Xiong J, Jiang L, Cui D, He J, Chen P, Li L, Wu C, Ma T, Shao S, Wang J, Yu D, Zhou B, Huang D, Schmitt CA, Tao R (2019) Epigenetic co-deregulation of EZH2/TET1 is a senescence-countering, actionable vulnerability in triple-negative breast cancer. Theranostics 9(3):761–777. https://doi.org/10.7150/thno.29520

    Article  CAS  Google Scholar 

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Acknowledgements

The authors thank the Director, Chittaranjan National Cancer Institute [CNCI], Kolkata, India for kind interest in the work. We would like to thank Mr. Balarko Chakraborty for his language editing help and valuable suggestions. We also thanks health professionals and pathology department for collection of specimens. Finally, authors specially thank all the 159 patients who participated in the study.

Funding

The financial support for this work DST-INSPIRE Fellowship grant Sanction order. No. IF170005/2017, EU-V dated 17.07.2017) to was provided by Mrs. P. Dutta, University Grants Commission-National Eligibility Test 46 Fellowship grant [Sr. No. 2061430780, Ref No.: 22/06/2014(i)EU-V to M. Basu and to NASI Senior Scientist Platinum Jubilee Fellowship (2020) awarded to Dr. C.K. Panda. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Department of Oncogene Regulation, Chittaranjan National Cancer Institute, 37, S.P. Mukherjee Road, Kolkata, West Bengal, 700026, India

    Priyanka Dutta, Mukta Basu & Chinmay Kumar Panda

  2. Department of Pathology, Nil Ratan Sircar Medical College and Hospital, Kolkata, 700014, India

    Anup Roy

  3. Department of Gynaecologic Oncology, Chittaranjan National Cancer Institute, 37, S.P. Mukherjee Road, Kolkata, West Bengal, 700026, India

    Ranajit Kumar Mandal

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  1. Priyanka Dutta
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  2. Mukta Basu
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  3. Anup Roy
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  4. Ranajit Kumar Mandal
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  5. Chinmay Kumar Panda
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Contributions

This work was conceptualized and manuscript was drafted by PD and CKP. PD performed the experiments in primary tumour and CACX cell lines while MB and RKM helped in interpreting the results and did the statistical analysis. RKM provided the tumour samples. AR was helpful in providing the pathological guidance and other resources.

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Correspondence to Chinmay Kumar Panda.

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All the authors of this study agreed to participate in this study.

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The authors are accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. Written informed consent was obtained from patients, as well as hospital authorities and institutional ethical committee prior to the study.

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FIGURE S1

: Pictorial representation of Laser Capture Microdissection procedure and downstream process in Hematoxylin- eosin-stained cervical epithelium (red line indicates the laser line separating the basal-parabasal layer from spinous layer of cervical epithelium tissue) (TIF 478 kb)

FIGURE S2

: Detection HPV in cervical tissue samples A. Representative image of HPV screening by MY09-MY11 primers in cervical tissue samples, (+ve control used as HPV 16 plasmid, -ve control used as distilled water) B. Representative image of HPV 16 prevalence in cervical tissue samples (+ve control used as HPV 16 plasmid, -ve control used as distilled water) C. Representative image of HPV 18 prevalence in cervical tissue samples (+ve control used as HPV 18 plasmid, -ve control used as distilled water). D. % of HPV prevalence in C.E (cervical epithelium), CIN and CACX samples. (TIF 587 kb)

10735_2023_10114_MOESM3_ESM.tif

FIGURE S3: Promoter Methylation analysis of DNMT1 and TET1 after treatment with different concentrations of 5-aza-dC doses (5 uM, 10 uM and 20 uM) in CACX cell lines (SiHa and HeLa); 1-2 (a,b); Representative images showing methylation status of DNMT1 & TET1, (K1 and K2- controls for DNA digestion and integrity respectively), (M=100 bp ladder, U.D=undigested DNA, D= Hpa II digested DNA) 3,4; Histogram representing quantitative methylation frequencies of DNMT1 and TET1 observed in CACX cell lines (* = significant p value < 0.05). (TIF 380 kb)

TABLE S1:

Flow chart of sample utilization. TABLE S2: Details of primers used for experiments. TABLE S3: Immuno-histochemical analysis of DNMT1, TET1 and HPV oncoprotein E7 in cervical epithelium and primary cervical lesions.TABLE S4: A: Co-relation of Expression of DNMT1 in basal-parabasal layer of HPV-ve cervical epithelium. B: Study of Co-relation between mRNA expression and protein expression of DNMT1, TET1 and E7 oncoprotein C: Co-relation of expression of E7 with DNMT1, TET1 in primary cervical lesions. D. Study of Co-relation between expression and methylation of DNMT1, TET1 in CACX samples. TABLE S5: Study of Co-relation between promoter methylation of DNMT1, TET1 between tissue DNA and plasma DNA of primary cervical lesions.TABLE S6: A: Study of Co-relation between overall alteration and protein expression of DNMT1 and TET1 in primary cervical lesions. B: Study of Co-relation between Overall Alteration of DNMT1, TET1 and E7 expression of primary cervical lesions. (DOCX 49 kb)

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Dutta, P., Basu, M., Roy, A. et al. High nuclear expression of DNMT1 in correlation with inactivation of TET1 portray worst prognosis among the cervical carcinoma patients: clinical implications. J Mol Histol 54, 89–102 (2023). https://doi.org/10.1007/s10735-023-10114-z

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