Cell-Free Circulating Tumor DNA Mutation Profiling for Cervical Carcinoma as Diagnostic Biomarker: A 50-Gene Module to Future Directive

  • H. B. GovardhanEmail author
  • I. A. Khaleel
  • S. A. Shubha
  • R. Manisha
  • S. Nivedita
  • N. Noopur
  • N. P. Jayashree
  • T. Fareena
  • K. Sweta
Original Article



Across the globe, cervical cancer is the most common female malignancy, second only to breast cancer, as in both incidence and mortality. Currently, tissue biopsy is the gold standard to verify carcinoma of uterine cervix initial diagnosis and can be challenging due to its invasive nature. In this study, our objective was a noninvasive genetic panel for timely detection of cervical carcinoma and its progression using cell-free tumor DNA (ctDNA).


Twenty-five cervical carcinoma patients were tested with a 50-gene tumor panel. ctDNA isolated from serum was checked for single-nucleotide variations (SNVs) or copy number alterations using targeted next-generation sequencing, with further validation of results by checking respective formalin-fixed paraffin-embedded tumor tissues for the same genetic alterations.


Out of 50 genes, 32 were detected in the serum samples. The SNVs detected included TP53 in 52.3% patients, CDKN2A in 47.6%, PTEN and STK11 in 33.3% patients, BRAF and VHL in 28.5% patients, EGFR and SMAD4 in 19% patients; CTNNB1, GNAS, KIT, APC, PIK3CA in 14.28% patients; SMARCB1, SMO, RET, FBXW7, ERBB2, CSF1R, CDH1, AKT1, ATM, EBB4, FGFR3, FLT3, HRAS, JAK3, MET, NOTCH1, NPM1, KRAS, PTPN11 in 4.7 to 9.5% patients. On combining alterations in BRAF, CDKN2A, EGFR, PIK3CA, PTEN, STK11, TP53 and VHL genes, at least one of the genetic alterations was found in 100% patients.


These findings illustrate that ctDNA is easily demonstrable and can be used as a surrogate for tissue biopsy in uterine cervix carcinoma.


Cervical carcinoma ctDNA Next-generation sequencing Minimally invasive Biomarker 



We acknowledge the contributions by Govardhan HB and Khaleel IA who conceived and designed the study protocol; Shubha S A who was responsible for acquisition of data; Manisha R for autonomously writing, reviewing and/or revising the manuscript; Nivedita S and Noopur N who provided technical support and study supervision; and Jayashree NP, Fareena T and Sweta K who gave help during the course of the study.

Compliance with Ethical Standards

Conflict of interest

The authors declare no conflicts of interest.

Ethical Approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.


  1. 1.
    Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D, Bray F. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN. Int J Cancer. 2012;136(2015):E359–86. Scholar
  2. 2.
    Chan KCA, Jiang P, Zheng YWL, Liao GJW, Sun H, Wong J, Siu SSN, Chan WC, Chan SL, Chan ATC, Lai PBS, Chiu RWK, Lo YMD. Cancer genome scanning in plasma: detection of tumor-associated copy number aberrations, single-nucleotide variants, and tumoral heterogeneity by massively parallel sequencing. Clin Chem. 2013;59:211–24. Scholar
  3. 3.
    Heitzer E, Ulz P, Belic J, Gutschi S, Quehenberger F, Fischereder K, Benezeder T, Auer M, Pischler C, Mannweiler S, Pichler M, Eisner F, Haeusler M, Riethdorf S, Pantel K, Samonigg H, Hoefler G, Augustin H, Geigl JB, Speicher MR. Tumor-associated copy number changes in the circulation of patients with prostate cancer identified through whole-genome sequencing. Genome Med. 2013;5:30. Scholar
  4. 4.
    Anker P, Mulcahy H, Chen XQ, Stroun M. Detection of circulating tumour DNA in the blood (plasma/serum) of cancer patients. Cancer Metastasis Rev. 1999;18:65–73.CrossRefGoogle Scholar
  5. 5.
    Lo YM, Zhang J, Leung TN, Lau TK, Chang AM, Hjelm NM. Rapid clearance of fetal DNA from maternal plasma. Am J Hum Genet. 1999;64:218–24. Scholar
  6. 6.
    De Mattos-Arruda L, Caldas C. Cell-free circulating tumour DNA as a liquid biopsy in breast cancer. Mol Oncol. 2016;10:464–74. Scholar
  7. 7.
    Murtaza M, Dawson S-J, Tsui DWY, Gale D, Forshew T, Piskorz AM, Parkinson C, Chin S-F, Kingsbury Z, Wong ASC, Marass F, Humphray S, Hadfield J, Bentley D, Chin TM, Brenton JD, Caldas C, Rosenfeld N. Non-invasive analysis of acquired resistance to cancer therapy by sequencing of plasma DNA. Nature. 2013;497:108–12. Scholar
  8. 8.
    Warton K, Mahon KL, Samimi G. Methylated circulating tumor DNA in blood: power in cancer prognosis and response. Endocr Relat Cancer. 2016;23:R157–71. Scholar
  9. 9.
    Diehl F, Schmidt K, Choti MA, Romans K, Goodman S, Li M, Thornton K, Agrawal N, Sokoll L, Szabo SA, Kinzler KW, Vogelstein B, Diaz LA Jr. Circulating mutant DNA to assess tumor dynamics. Nat Med. 2008;14:985–90. Scholar
  10. 10.
    Thierry AR, Mouliere F, El Messaoudi S, Mollevi C, Lopez-Crapez E, Rolet F, Gillet B, Gongora C, Dechelotte P, Robert B, Del Rio M, Lamy PJ, Bibeau F, Nouaille M, Loriot V, Jarrousse AS, Molina F, Mathonnet M, Pezet D, Ychou M. Clinical validation of the detection of KRAS and BRAF mutations from circulating tumor DNA. Nat Med. 2014;20:430–5. Scholar
  11. 11.
    Freidin MB, Freydina DV, Leung M, Montero Fernandez A, Nicholson AG, Lim E. Circulating tumor DNA outperforms circulating tumor cells for KRAS mutation detection in thoracic malignancies. Clin Chem. 2015;61:1299–304. Scholar
  12. 12.
    Forshew T, Murtaza M, Parkinson C, Gale D, Tsui DW, Kaper F, et al. Noninvasive identification and monitoring of cancer mutations by targeted deep sequencing of plasma DNA. Sci Transl Med. 2012;4(136):136ra68.CrossRefGoogle Scholar
  13. 13.
    Newman AM, Bratman SV, To J, Wynne JF, Eclov NCW, Modlin LA, Liu CL, Neal JW, Wakelee HA, Merritt RE, Shrager JB, Loo BW, Alizadeh AA, Diehn M. An ultrasensitive method for quantitating circulating tumor DNA with broad patient coverage. Nat Med. 2014;20:548–54. Scholar
  14. 14.
    Jones S, Anagnostou V, Lytle K, Parpart-Li S, Nesselbush M, Riley DR, et al. Personalized genomic analyses for cancer mutation discovery and interpretation. Sci Transl Med. 2015;7(283):283ra53.CrossRefGoogle Scholar
  15. 15.
    Schwaederle M, Husain H, Fanta PT, Piccioni DE, Kesari S, Schwab RB, Banks KC, Lanman RB, Talasaz A, Parker BA, Kurzrock R. Detection rate of actionable mutations in diverse cancers using a biopsy-free (blood) circulating tumor cell DNA assay. Oncotarget. 2016;7:9707–17. Scholar
  16. 16.
    Frenel JS, Carreira S, Goodall J, Roda D, Perez-Lopez R, Tunariu N et al. Serial next-generation sequencing of circulating cell-free DNA evaluating tumor clone response to molecularly targeted drug administration. Clin Cancer Res. 2015;21(20):4586–96. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Association of Gynecologic Oncologists of India 2018

Authors and Affiliations

  1. 1.Department of Radiation OncologyKidwai Cancer InstituteBangaloreIndia

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