Altered mitochondrial DNA copy number in cervical exfoliated cells among high‑risk HPV‑positive and HPV‑negative women

Al‑Awadhi,Rana; Alroomy,Moody; Al‑Waheeb,Salah; Alwehaidah,Materah Salem

doi:10.3892/etm.2023.12220

Altered mitochondrial DNA copy number in cervical exfoliated cells among high‑risk HPV‑positive and HPV‑negative women

Authors:
- Rana Al‑Awadhi
- Moody Alroomy
- Salah Al‑Waheeb
- Materah Salem Alwehaidah
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Affiliations: Department of Medical Laboratory Sciences, Faculty of Allied Health Sciences, Kuwait University, Jabriyah 90805, Kuwait, Cytology Laboratory, Maternity Hospital and Mubarak Al‑Kabeer Hospital, Ministry of Health, Kuwait University, Jabriyah 90805, Kuwait
Published online on: September 21, 2023 https://doi.org/10.3892/etm.2023.12220
Article Number: 521

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Abstract

The majority of cervical cancer cases are due to human papillomavirus (HPV) infection. However, certain cases of cervical cancer are not caused by HPV. Recent studies have shown a link between altered mitochondrial DNA (mtDNA) copy number, an indicative measure of mitochondrial dysfunction, and cervical cancer in women who test positive for HPV. However, the role of the mtDNA copy number in HPV‑negative cervical cancer has remained elusive. In the present study, the mtDNA copy number was determined using quantitative PCR as the ratio between mtDNA and nuclear DNA in 287 ThinPrep cervical samples, including 143 cases with cervical abnormalities and 144 control subjects with high‑risk (hr)‑HPV positive or HPV‑negative status. In an overall analysis of cases categorized based on the cytology diagnosis into squamous cervical carcinoma/high‑grade squamous intraepithelial lesions (SCC/HSIL), low‑grade squamous intraepithelial lesions (LSIL) and normal controls, the mtDNA copy number was significantly higher in all cases compared to the controls and a higher mtDNA copy number was observed in SCC/HSIL compared to LSIL cases. In the stratification analyses based on hr‑HPV positive and HPV‑negative status, an increased mtDNA copy number was observed in the cases compared with the controls regardless of their HPV status (P<0.05). When cases with cervical abnormalities were categorized based on histological diagnosis into cervical intraepithelial neoplasia (CIN)2/CIN3 and CIN1, an overall analysis indicated an increased mtDNA copy number in CIN2/CIN3 compared to CIN1 (P=0.01). Stratification analyses of these cases based on HPV status revealed a higher mtDNA copy number in CIN2/CIN3 compared to CIN1 regardless of HPV infection (P<0.05). These results showed that an elevated mtDNA copy number in subjects with cervical abnormalities was not influenced by the HPV status and suggested the possibility of its role in the progression of cervical cancer. The increased mtDNA copy number may be an adaptive response mechanism to compensate for mtDNA oxidative stress and energy deficiency, possibly induced by HPV infection and other environmental exposures.

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1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015.PubMed/NCBI View Article : Google Scholar
2	Mattiuzzi C and Lippi G: Cancer statistics: A comparison between World Health Organization (WHO) and Global Burden of Disease (GBD). Eur J Public Health. 30:1026–1027. 2020.PubMed/NCBI View Article : Google Scholar
3	Alkhalawi E, Al-Madouj A and Al-Zahrani A: Cervical cancer incidence and trends among nationals of the gulf cooperation council states, 1998-2012. Gulf J Oncolog. 1:7–13. 2019.PubMed/NCBI
4	zur Hausen H: Papillomaviruses and cancer: From basic studies to clinical application. Nat Rev Cancer. 2:342–350. 2002.PubMed/NCBI View Article : Google Scholar
5	Zhang S, Xu H, Zhang L and Qiao Y: Cervical cancer: Epidemiology, risk factors and screening. Chin J Cancer Res. 32:720–728. 2020.PubMed/NCBI View Article : Google Scholar
6	Roura E, Castellsague X, Pawlita M, Travier N, Waterboer T, Margall N, Bosch FX, de Sanjosé S, Dillner J, Gram IT, et al: Smoking as a major risk factor for cervical cancer and pre-cancer: Results from the EPIC cohort. Int J Cancer. 135:453–466. 2014.PubMed/NCBI View Article : Google Scholar
7	Gissmann L and zur Hausen H: Partial characterization of viral DNA from human genital warts (Condylomata acuminata). Int J Cancer. 25:605–609. 1980.PubMed/NCBI View Article : Google Scholar
8	Schiffman M, Castle PE, Jeronimo J, Rodriguez AC and Wacholder S: Human papillomavirus and cervical cancer. Lancet. 370:890–907. 2007.PubMed/NCBI View Article : Google Scholar
9	Bloem P and Ogbuanu I: Vaccination to prevent human papillomavirus infections: From promise to practice. PLoS Med. 14(e1002325)2017.PubMed/NCBI View Article : Google Scholar
10	US Preventive Services Task Force. Curry SJ, Krist AH, Owens DK, Barry MJ, Caughey AB, Davidson KW, Doubeni CA, Epling JW Jr, Kemper AR, et al: Screening for CERVICAL CANcer: US preventive services task force recommendation statement. JAMA. 320:674–686. 2018.PubMed/NCBI View Article : Google Scholar
11	Delpero E and Selk A: Shifting from cytology to HPV testing for cervical cancer screening in Canada. CMAJ. 194:E613–E615. 2022.PubMed/NCBI View Article : Google Scholar
12	Aitken CA, van Agt HME, Siebers AG, van Kemenade FJ, Niesters HGM, Melchers WJG, Vedder JEM, Schuurman R, van den Brule AJC, van der Linden HC, et al: Introduction of primary screening using high-risk HPV DNA detection in the Dutch cervical cancer screening programme: A population-based cohort study. BMC Med. 17(228)2019.PubMed/NCBI View Article : Google Scholar
13	Blatt AJ, Kennedy R, Luff RD, Austin RM and Rabin DS: Comparison of cervical cancer screening results among 256,648 women in multiple clinical practices. Cancer Cytopathol. 123:282–288. 2015.PubMed/NCBI View Article : Google Scholar
14	Petry KU, Liebrich C, Luyten A, Zander M and Iftner T: Surgical staging identified false HPV-negative cases in a large series of invasive cervical cancers. Papillomavirus Res. 4:85–89. 2017.PubMed/NCBI View Article : Google Scholar
15	Clifford GM, Smith JS, Plummer M, Munoz N and Franceschi S: Human papillomavirus types in invasive cervical cancer worldwide: A meta-analysis. Br J Cancer. 88:63–73. 2003.PubMed/NCBI View Article : Google Scholar
16	Guan P, Howell-Jones R, Li N, Bruni L, de Sanjosé S, Franceschi S and Clifford GM: Human papillomavirus types in 115,789 HPV-positive women: A meta-analysis from cervical infection to cancer. Int J Cancer. 131:2349–2359. 2012.PubMed/NCBI View Article : Google Scholar
17	Hohn AK, Brambs CE, Hiller GGR, May D, Schmoeckel E and Horn LC: 2020 WHO classification of female genital tumors. Geburtshilfe Frauenheilkd. 81:1145–1153. 2021.PubMed/NCBI View Article : Google Scholar
18	Arezzo F, Cormio G, Loizzi V, Cazzato G, Cataldo V, Lombardi C, Ingravallo G, Resta L and Cicinelli E: HPV-Negative cervical cancer: A narrative review. Diagnostics (Basel). 11(952)2021.PubMed/NCBI View Article : Google Scholar
19	Fernandes A, Viveros-Carreno D, Hoegl J, Avila M and Pareja R: Human papillomavirus-independent cervical cancer. Int J Gynecol Cancer. 32:1–7. 2022.PubMed/NCBI View Article : Google Scholar
20	Yusoff AAM, Abdullah WSW, Khair SZNM and Radzak SMA: A comprehensive overview of mitochondrial DNA 4977-bp deletion in cancer studies. Oncol Rev. 13(409)2019.PubMed/NCBI View Article : Google Scholar
21	Ksiezakowska-Lakoma K, Zyla M and Wilczynski JR: Mitochondrial dysfunction in cancer. Prz Menopauzalny. 13:136–144. 2014.PubMed/NCBI View Article : Google Scholar
22	Kabekkodu SP, Bhat S, Mascarenhas R, Mallya S, Bhat M, Pandey D, Kushtagi P, Thangaraj K, Gopinath PM and Satyamoorthy K: Mitochondrial DNA variation analysis in cervical cancer. Mitochondrion. 16:73–82. 2014.PubMed/NCBI View Article : Google Scholar
23	Warowicka A, Kwasniewska A and Gozdzicka-Jozefiak A: Alterations in mtDNA: A qualitative and quantitative study associated with cervical cancer development. Gynecol Oncol. 129:193–198. 2013.PubMed/NCBI View Article : Google Scholar
24	Zhai K, Chang L, Zhang Q, Liu B and Wu Y: Mitochondrial C150T polymorphism increases the risk of cervical cancer and HPV infection. Mitochondrion. 11:559–563. 2011.PubMed/NCBI View Article : Google Scholar
25	DiMauro S and Wallace DC: Mitochondrial DNA in Human Pathology. Raven Press, New York, NY, 1993.
26	Clay Montier LL, Deng JJ and Bai Y: Number matters: Control of mammalian mitochondrial DNA copy number. J Genet Genomics. 36:125–131. 2009.PubMed/NCBI View Article : Google Scholar
27	Lee HC and Wei YH: Mitochondrial biogenesis and mitochondrial DNA maintenance of mammalian cells under oxidative stress. Int J Biochem Cell Biol. 37:822–834. 2005.PubMed/NCBI View Article : Google Scholar
28	Lee HC, Yin PH, Lu CY, Chi CW and Wei YH: Increase of mitochondria and mitochondrial DNA in response to oxidative stress in human cells. Biochem J. 348:425–432. 2000.PubMed/NCBI
29	Lee JW, Park KD, Im JA, Kim MY and Lee DC: Mitochondrial DNA copy number in peripheral blood is associated with cognitive function in apparently healthy elderly women. Clin Chim Acta. 411:592–596. 2010.PubMed/NCBI View Article : Google Scholar
30	Pyle A, Anugrha H, Kurzawa-Akanbi M, Yarnall A, Burn D and Hudson G: Reduced mitochondrial DNA copy number is a biomarker of Parkinson's disease. Neurobiol Aging. 38:216 e7–216 e10. 2016.PubMed/NCBI View Article : Google Scholar
31	Svendsen AJ, Tan Q, Jakobsen MA, Thyagarajan B, Nygaard M, Christiansen L and Mengel-From J: White blood cell mitochondrial DNA copy number is decreased in rheumatoid arthritis and linked with risk factors. A twin study. J Autoimmun. 96:142–146. 2019.PubMed/NCBI View Article : Google Scholar
32	Reznik E, Miller ML, Senbabaoglu Y, Riaz N, Sarungbam J, Tickoo SK, Al-Ahmadie HA, Lee W, Seshan VE, Hakimi AA and Sander C: Mitochondrial DNA copy number variation across human cancers. Elife. 5(e10769)2016.PubMed/NCBI View Article : Google Scholar
33	Hu L, Yao X and Shen Y: Altered mitochondrial DNA copy number contributes to human cancer risk: Evidence from an updated meta-analysis. Sci Rep. 6(35859)2016.PubMed/NCBI View Article : Google Scholar
34	Sun W, Qin X, Zhou J, Xu M, Lyu Z, Li X, Zhang K, Dai M, Li N and Hang D: Mitochondrial DNA copy number in cervical exfoliated cells and risk of cervical cancer among HPV-positive women. BMC Womens Health. 20(139)2020.PubMed/NCBI View Article : Google Scholar
35	Nayar R and Wilbur DC: The Pap test and Bethesda 2014. Cancer Cytopathol. 123:271–281. 2015.PubMed/NCBI View Article : Google Scholar
36	Al-Awadhi R, Chehadeh W and Kapila K: Prevalence of human papillomavirus among women with normal cervical cytology in Kuwait. J Med Virol. 83:453–460. 2011.PubMed/NCBI View Article : Google Scholar
37	Alwehaidah MS, AlFadhli S and Al-Kafaji G: Leukocyte mitochondrial DNA copy number is a potential non-invasive biomarker for psoriasis. PLoS One. 17(e0270714)2022.PubMed/NCBI View Article : Google Scholar
38	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
39	Filograna R, Mennuni M, Alsina D and Larsson NG: Mitochondrial DNA copy number in human disease: The more the better? FEBS Lett. 595:976–1002. 2021.PubMed/NCBI View Article : Google Scholar
40	Al-Kafaji G, Bakheit HF, Alharbi MA, Farahat AA, Jailani M, Ebrahin BH and Bakhiet M: Mitochondrial DNA copy number in peripheral blood as a potential non-invasive biomarker for multiple sclerosis. Neuromolecular Med. 22:304–313. 2020.PubMed/NCBI View Article : Google Scholar
41	Abd Radzak SM, Mohd Khair SZN, Ahmad F, Patar A, Idris Z and Mohamed Yusoff AA: Insights regarding mitochondrial DNA copy number alterations in human cancer (Review). Int J Mol Med. 50(104)2022.PubMed/NCBI View Article : Google Scholar
42	Mei H, Sun S, Bai Y, Chen Y, Chai R and Li H: Reduced mtDNA copy number increases the sensitivity of tumor cells to chemotherapeutic drugs. Cell Death Dis. 6(e1710)2015.PubMed/NCBI View Article : Google Scholar
43	Woodman CB, Collins SI and Young LS: The natural history of cervical HPV infection: Unresolved issues. Nat Rev Cancer. 7:11–22. 2007.PubMed/NCBI View Article : Google Scholar
44	Bedell SL, Goldstein LS, Goldstein AR and Goldstein AT: Cervical cancer screening: Past, present, and future. Sex Med Rev. 8:28–37. 2020.PubMed/NCBI View Article : Google Scholar
45	Williams VM, Filippova M, Filippov V, Payne KJ and Duerksen-Hughes P: Human papillomavirus type 16 E6* induces oxidative stress and DNA damage. J Virol. 88:6751–6761. 2014.PubMed/NCBI View Article : Google Scholar
46	Marullo R, Werner E, Zhang H, Chen GZ, Shin DM and Doetsch PW: HPV16 E6 and E7 proteins induce a chronic oxidative stress response via NOX2 that causes genomic instability and increased susceptibility to DNA damage in head and neck cancer cells. Carcinogenesis. 36:1397–1406. 2015.PubMed/NCBI View Article : Google Scholar
47	Buckley CH, Butler EB and Fox H: Cervical intraepithelial neoplasia. J Clin Pathol. 35:1–13. 1982.PubMed/NCBI View Article : Google Scholar
48	Leeman A, Del Pino M, Marimon L, Torné A, Ordi J, Ter Harmsel B, Meijer CJLM, Jenkins D, Van Kemenade FJ and Quint WGV: Reliable identification of women with CIN3+ using hrHPV genotyping and methylation markers in a cytology-screened referral population. Int J Cancer. 144:160–168. 2019.PubMed/NCBI View Article : Google Scholar
49	Coquillard G, Palao B and Patterson BK: Quantification of intracellular HPV E6/E7 mRNA expression increases the specificity and positive predictive value of cervical cancer screening compared to HPV DNA. Gynecol Oncol. 120:89–93. 2011.PubMed/NCBI View Article : Google Scholar
50	Rai NK, Panjwani G, Ghosh AK, Haque R and Sharma LK: Analysis of mitochondrial DNA copy number variation in blood and tissue samples of metastatic breast cancer patients (A pilot study). Biochem Biophys Rep. 26(100931)2021.PubMed/NCBI View Article : Google Scholar
51	Chen N, Wen S, Sun X, Fang Q, Huang L, Liu S, Li W and Qiu M: Elevated Mitochondrial DNA copy number in peripheral blood and tissue predict the opposite outcome of cancer: A meta-analysis. Sci Rep. 6(37404)2016.PubMed/NCBI View Article : Google Scholar