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Quantification of Circulating Free DNA as a Diagnostic Marker in Gall Bladder Cancer

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Pathology & Oncology Research

Abstract

Gall bladder Carcinoma (GBC) is the fifth most common cancer of the digestive tract and frequently diagnosed in late stage of disease. Estimation of circulating free DNA (cfDNA) in serum has been applied as a “liquid biopsy” in several deep seated malignancies. Its value in diagnosis of gall bladder carcinoma has not been studied. The present study was designed to assess the role of cfDNA in the diagnosis of GBC and correlate levels with the TNM stage. Serum was collected from 34 patients with GBC and 39 age and sex matched controls including 22 cholecystitis and 17 healthy individuals. Serum cfDNA levels were measured through quantitative polymerase chain reaction (qPCR) by amplification of β-globin gene. Performance of the assay was calculated through the receiver operating characteristic (ROC) curve. The cfDNA level was significantly lower in healthy controls and cholecystitis (89.32 ± 59.76 ng/ml, 174.21 ± 99.93 ng/ml) compared to GBC (1245.91 ± 892.46 ng/ml, p = <0.001). The cfDNA level was significantly associated with TNM stage, lymph node involvement and jaundice (0.002, 0.027, and 0.041, respectively). Area under curve of ROC analysis for cancer group versus healthy and cholecystitis group was 1.00 and 0.983 with sensitivity of 100 %, 88.24 % and specificity of 100 % respectively. Quantitative analysis of cfDNA may distinguish cholecystitis and gall bladder carcinoma and may serve as new diagnostic, noninvasive marker adjunct to imaging for the diagnosis of GBC.

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References

  1. Mishra S, Chaturvedi A, Mishra NC, Sharma ID (2013) Carcinoma of the gallbladder. The Lancet Oncology 4:167–176

    Article  Google Scholar 

  2. Hamdani NS, Qadri SK, Aggarwalla R, Bhartia VK, Chaudhuri S, Debakshi S, Baig SJ, Pal NK (2013) Clinicopathological Study of Gall Bladder Carcinoma with Special Reference to Gallstones: Our 8-year Experience from Eastern India. Asian Pacific J Cancer Prev 13(11):5613–5617

    Article  Google Scholar 

  3. Goldin RD, Roa JC (2009) Gallbladder cancer: a morphological and molecular update. Histopathology 55:218–229

    Article  PubMed  Google Scholar 

  4. Kapoor VK, Michael AJ Mc (2003) Gallbladder cancer: An ‘Indian’ disease. Natl Med J India 4:16.

  5. Peng HH, Zhang YD, Gong LS, Liu WD, Zhang Y (2013) Increased expression of microRNA-335 predicts a favorable prognosis in primary gallbladder carcinoma. Onco Targets Ther 6:1625–1630.

  6. Liu TY, Tan ZJ, Jiang L, JF G, XS W, Cao Y, Li ML, KJ W, Liu YB (2013) Curcumin induces apoptosis in gallbladder carcinoma cell line GBC-SD cells. Cancer Cell Int 13:6

    Article  Google Scholar 

  7. Stroun M, Maurice P, Vasioukhin V, Lyautey J, Lederry C, Lefort F (2000) The origin and mechanism of circulating DNA. Ann N Y Acad Sci 906:161–168

    Article  CAS  PubMed  Google Scholar 

  8. Ellinger J, Bastian PJ, Haan KI, Heukamp LC, Buettner R, Fimmers R, Mueller SC, Von Ruecker A (2008) Noncancerous PTGS2 DNA fragments of apoptotic origin in sera of prostate cancer patients qualify as diagnostic and prognostic indicators. Int J Cancer 122(1):138–143

    Article  CAS  PubMed  Google Scholar 

  9. Sozzi G, Conte D, Mariani L (2001) Analysis of circulating tumor DNA in plasma at diagnosis and during follow-up of lung cancer patients. Cancer Res 61:4675–4678

    CAS  PubMed  Google Scholar 

  10. Umetani N, Kim J, Hiramatsu S (2006) Increased integrity of free circulating DNA in sera of patients with colorectal or periampullary cancer: direct quantitative PCR for ALU repeats. Clin Chem 52:1062–1069.

  11. Trejo-Becerril C, Perez-Cardenas E, Trevino-Cuevas H (2003) Circulating nucleosomes and response to chemotherapy: an in vitro, in vivo and clinical study on cervical cancer patients. Int J Cancer 104:663–668

    Article  CAS  PubMed  Google Scholar 

  12. Chang HW, Lee SM, Goodman SN (2002) Assessment of plasma DNA levels, allelic imbalance, and CA 125 as diagnostic tests for cancer. J Natl. Cancer Inst 94:1697–1703

    Article  CAS  PubMed  Google Scholar 

  13. Gal S, Fidler C, Lo YMD, Taylor M, Han C, Moore J, Harris AL, Wainscoat JS (2004) Quantitation of circulating DNA in the serum of breast cancer patients by real-time PCR. Br J Cancer 90:1211–1215

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Ellinger J, Wittkamp V, Albers P (2009) Cell-free circulating DNA: diagnostic value in patients with testicular germ cell cancer. J Urol 181:363–371

    Article  CAS  PubMed  Google Scholar 

  15. Ellinger J, Bastian PJ, Ellinger N (2008) Apoptotic DNA fragments in serum of patients with muscle invasive bladder cancer: a prognostic entity. Cancer Lett 264:274–280

    Article  CAS  PubMed  Google Scholar 

  16. Altimari A, Grigioni AD, Benedettini E (2008) Diagnostic role of circulating free plasma DNA detection in patients with localized prostate cancer. Am. J Clin Pathol 129:756–762

    Article  CAS  PubMed  Google Scholar 

  17. Sai S, Ichikawa D, Tomita H, Ikoma D, Tani N, Ikoma H, Kikuchi S, Fujiwara H, Ueda Y, Otsuji E (2007) Quantification of Plasma Cell-free DNA in Patients with Gastric Cancer. Anticancer Res 27:2747–2752

    CAS  PubMed  Google Scholar 

  18. Chen K, Zhang H, Zhang L, SQ J, Qi J, Huang DF, Li F, Wei Q, Jing Zhang J (2013) Value of circulating cell-free DNA in diagnosis of hepatocellular carcinoma. World J Gastroenterol 19(20):3143–3149

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Lo YMD, Rainer TH, Chan LYS, Hjelm NM, Cocks RA (2000) Plasma DNA as a Prognostic Marker in Trauma Patients. Clin Chem 46(3):319–323

    CAS  PubMed  Google Scholar 

  20. Ghosh M, Sakhuja P, Singh S, Agarwal AK (2013) p53 and beta catenin expression in gallbladder tissues and correlation with tumor progression in gallbladder cancer. Saudi J Gastroenterol 19:34–39

    Article  PubMed  PubMed Central  Google Scholar 

  21. He CZ, Zhang KH, Li Q, Liu XH, Hong Y, Lv NH (2013) Combined use of AFP, CEA, CA125 and CAl9–9 improves the sensitivity for the diagnosis of gastric cancer. BMC Gastroenterol 13:87.

  22. Zhang D, Yu M, Xu T, Xiong B (2013) Predictive value of serum CEA, CA19–9 and CA125 in diagnosis of colorectal liver metastasis in Chinese population. Hepato-Gastroenterology 60:1297–1301

    CAS  PubMed  Google Scholar 

  23. Nath A, Kumar S, Singh M, Nadeem T, Murti K, Gupta AK, Das P, Kumar A (2015) Combined assessment of lipid peroxidation, Cea and Ca19.9 biomarkers in advanced stage gall bladder cancer. Int J Recent Sci Res 6::3136–3314

    Google Scholar 

  24. Leon SA, Shapiro B, Sklaroff DM, Yaros MJ (1977) Free DNA in the serum of cancer patients and the effect of therapy. Cancer Res 37:646–650

    CAS  PubMed  Google Scholar 

  25. Umetani N, Hiramatsu S, Hoon DSB (2006) Higher amount of free circulating DNA in serum than in plasma is not mainly caused by contaminated extraneous DNA during separation. Annals NY Acad Sci 1075:299–307

    Article  CAS  Google Scholar 

  26. Gormally E, Caboux E, Vineis P, Hainaut P (2007) Circulating free DNA in plasma or serum as biomarker of carcinogenesis: Practical aspects and biological significance. Mutat Res 635:105–117

    Article  CAS  PubMed  Google Scholar 

  27. Pinzani P, Salvianti F, Pazzagli M, Orlando C (2010) Circulating nucleic acids in cancer and pregnancy. Methods 50:302–307

    Article  CAS  PubMed  Google Scholar 

  28. Swarup V, Rajeswari MR (2007) Circulating (cell-free) nucleic acids – a promising, Non-invasive tool for early detection of several human diseases. FEBS Lett 581:795–799

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

Authors wish to acknowledge and thanks Department of Science and Technology (DST), New Delhi, India for providing INSPIRE Research Fellowship (IF120759) support to Swati Kumari.

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Correspondence to Nuzhat Husain.

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Kumari, S., Tewari, S., Husain, N. et al. Quantification of Circulating Free DNA as a Diagnostic Marker in Gall Bladder Cancer. Pathol. Oncol. Res. 23, 91–97 (2017). https://doi.org/10.1007/s12253-016-0087-0

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  • DOI: https://doi.org/10.1007/s12253-016-0087-0

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