Breast Cancer Research and Treatment

, Volume 108, Issue 3, pp 427–434 | Cite as

Quantitative analysis of mitochondrial DNA 4977-bp deletion in sporadic breast cancer and benign breast diseases

  • Chuanzhong Ye
  • Xiao-Ou Shu
  • Wanqing Wen
  • Larry Pierce
  • Regina Courtney
  • Yu-Tang Gao
  • Wei Zheng
  • Qiuyin Cai
Epidemiology

Abstract

The mitochondrial DNA (mtDNA) 4977-bp deletion (ΔmtDNA4977 mutation) is one of the most frequently observed mtDNA mutations in human tissues and may play a role in carcinogenesis. Only a few studies have evaluated ΔmtDNA4977 mutation in breast cancer tissue, and the findings have been inconsistent, which may be due to methodological differences. In this study, we developed a quantitative real-time PCR assay to assess the level of the ΔmtDNA4977 mutation in tumor tissue samples from 55 primary breast cancer patients and 21 patients with benign breast disease (BBD). The ΔmtDNA4977 mutation was detected in all of the samples with levels varying from 0.000149% to 7.0%. The ΔmtDNA4977 mutation levels were lower in tumor tissues than in adjacent normal tissues in both breast cancer and BBD subjects. The differences, however, were not statistically significant. No significant difference between breast cancer and BBD patients was found in the ΔmtDNA4977 mutation levels of tumor tissues and adjacent normal tissues. The ΔmtDNA4977 mutation levels were not significantly associated with clinicopathological characteristics (age, histology, tumor stage, and ER/PR status) in breast cancer or BBD patients. These results do not support the notion that the mitochondrial DNA 4977-bp deletion plays a major role in breast carcinogenesis.

Keywords

Benign breast disease Breast cancer mtDNA 4977-bp deletion Quantitative real-time PCR assay 

Notes

Acknowledgments

We thank Dr. Konrad Huppi (Cancer Prevention Studies Branch, Center for Cancer Research, National Cancer Institute, Bethesda, MD) for kindly providing mitochondrial DNA samples, Bethanie Hull and Brandy Venuti for technical assistance in the preparation of this article, and all of the study participants and research staff of the Shanghai Breast Cancer Study for their support. This research was supported by U.S. Department of Defense grant DAMD17-02-1-0603 and National Cancer Institute grant R01 CA064277.

References

  1. 1.
    Zhu W, Qin W, Bradley P, Wessel A, Puckett CL, Sauter ER (2005) Mitochondrial DNA mutations in breast cancer tissue and in matched nipple aspirate fluid. Carcinogenesis 26:145–152PubMedCrossRefGoogle Scholar
  2. 2.
    Andrews RM, Kubacka I, Chinnery PF, Lightowlers RN, Turnbull DM, Howell N (1999) Reanalysis and revision of the Cambridge reference sequence for human mitochondrial DNA. Nat Genet 23:147PubMedCrossRefGoogle Scholar
  3. 3.
    Anderson S, Bankier AT, Barrell BG, de Bruijn MH, Coulson AR, Drouin J, Eperon IC, Nierlich DP, Roe BA, Sanger F, Schreier PH, Smith AJ, Staden R, Young IG (1981) Sequence and organization of the human mitochondrial genome. Nature 290:457–465PubMedCrossRefGoogle Scholar
  4. 4.
    Dani MA, Dani SU, Lima SP, Martinez A, Rossi BM, Soares F, Zago MA, Simpson AJ (2004) Less Deltamt DNA4977 than normal in various types of tumors suggests that cancer cells are essentially free of this mutation. Genet Mol Res 3:395–409PubMedGoogle Scholar
  5. 5.
    Tan DJ, Chang J, Liu LL, Bai RK, Wang YF, Yeh KT, Wong LJ (2006) Significance of somatic mutations and content alteration of mitochondrial DNA in esophageal cancer. BMC Cancer 6:93PubMedCrossRefGoogle Scholar
  6. 6.
    Roy D, Liehr JG (1999) Estrogen, DNA damage and mutations. Mutat Res 424:107–115PubMedGoogle Scholar
  7. 7.
    Bianchi MS, Bianchi NO, Bailliet G (1995) Mitochondrial DNA mutations in normal and tumor tissues from breast cancer patients. Cytogenet Cell Genet 71:99–103PubMedGoogle Scholar
  8. 8.
    Tseng LM, Yin PH, Chi CW, Hsu CY, Wu CW, Lee LM, Wei YH, Lee HC (2006) Mitochondrial DNA mutations and mitochondrial DNA depletion in breast cancer. Genes Chromosomes Cancer 45:629–638PubMedCrossRefGoogle Scholar
  9. 9.
    Zhu W, Qin W, Sauter ER (2004) Large-scale mitochondrial DNA deletion mutations and nuclear genome instability in human breast cancer. Cancer Detect Prev 28:119–126PubMedCrossRefGoogle Scholar
  10. 10.
    Gao YT, Shu XO, Dai Q, Potter JD, Brinton LA, Wen W, Sellers TA, Kushi LH, Ruan Z, Bostick RM, Jin F, Zheng W (2000) Association of menstrual and reproductive factors with breast cancer risk: results from the Shanghai Breast Cancer Study. Int J Cancer 87:295–300PubMedCrossRefGoogle Scholar
  11. 11.
    Schnitt S, Connolly J (1999) Pathology of benign breast disorders In: Harris JR, Lippman ME, Morrow M, Osborne CK (eds) Diseases of the breast. Lippincott Williams & Wilkins, Philadelphia, pp 75–93Google Scholar
  12. 12.
    Levin BC, Holland KA, Hancock DK, Coble M, Parsons TJ, Kienker LJ, Williams DW, Jones M, Richie KL (2003) Comparison of the complete mtDNA genome sequences of human cell lines—HL-60 and GM10742A—from individuals with pro-myelocytic leukemia and leber hereditary optic neuropathy, respectively, and the inclusion of HL-60 in the NIST human mitochondrial DNA standard reference material—SRM 2392-I. Mitochondrion 2:387–400PubMedCrossRefGoogle Scholar
  13. 13.
    Hirakawa M, Tanaka T, Hashimoto Y, Kuroda M, Takagi T, Nakamura Y (2002) JSNP: a database of common gene variations in the Japanese population. Nucleic Acids Res 30:158–162PubMedCrossRefGoogle Scholar
  14. 14.
    Smigielski EM, Sirotkin K, Ward M, Sherry ST (2000) dbSNP: a database of single nucleotide polymorphisms. Nucleic Acids Res 28:352–355PubMedCrossRefGoogle Scholar
  15. 15.
    Pushnova EA, Geier M, Zhu YS (2000) An easy and accurate agarose gel assay for quantitation of bacterial plasmid copy numbers. Anal Biochem 284:70–76PubMedCrossRefGoogle Scholar
  16. 16.
    Chuanzhong Y, Ming G, Fanglin Z, Haijiao C, Zhen L, Shiping C, Yong Kang Z (2002) Real-time quantitative reverse transcription-PCR assay for renal cell carcinoma-associated antigen G250. Clin Chim Acta 318:33–40PubMedCrossRefGoogle Scholar
  17. 17.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  18. 18.
    Shieh DB, Chou WP, Wei YH, Wong TY, Jin YT (2004) Mitochondrial DNA 4,977-bp deletion in paired oral cancer and precancerous lesions revealed by laser microdissection and real-time quantitative PCR. Ann NY Acad Sci 1011:154–167PubMedCrossRefGoogle Scholar
  19. 19.
    Pogozelski WK, Hamel CJ, Woeller CF, Jackson WE, Zullo SJ, Fischel-Ghodsian N, Blakely WF (2003) Quantification of total mitochondrial DNA and the 4977-bp common deletion in Pearson’s syndrome lymphoblasts using a fluorogenic 5′-nuclease (TaqMan) real-time polymerase chain reaction assay and plasmid external calibration standards. Mitochondrion 2:415–427PubMedCrossRefGoogle Scholar
  20. 20.
    He L, Chinnery PF, Durham SE, Blakely EL, Wardell TM, Borthwick GM, Taylor RW, Turnbull DM (2002) Detection and quantification of mitochondrial DNA deletions in individual cells by real-time PCR. Nucleic Acids Res 30:e68PubMedCrossRefGoogle Scholar
  21. 21.
    Nicklas JA, Brooks EM, Hunter TC, Single R, Branda RF (2004) Development of a quantitative PCR (TaqMan) assay for relative mitochondrial DNA copy number and the common mitochondrial DNA deletion in the rat. Environ Mol Mutagen 44:313–320PubMedCrossRefGoogle Scholar
  22. 22.
    Kakiuchi C, Ishiwata M, Kametani M, Nelson C, Iwamoto K, Kato T (2005) Quantitative analysis of mitochondrial DNA deletions in the brains of patients with bipolar disorder and schizophrenia. Int J Neuropsychopharmacol 8:515–522PubMedCrossRefGoogle Scholar
  23. 23.
    Porteous WK, James AM, Sheard PW, Porteous CM, Packer MA, Hyslop SJ, Melton JV, Pang CY, Wei YH, Murphy MP (1998) Bioenergetic consequences of accumulating the common 4977-bp mitochondrial DNA deletion. Eur J Biochem 257:192–201PubMedCrossRefGoogle Scholar
  24. 24.
    Schoeler S, Szibor R, Gellerich FN, Wartmann T, Mawrin C, Dietzmann K, Kirches E (2005) Mitochondrial DNA deletions sensitize cells to apoptosis at low heteroplasmy levels. Biochem Biophys Res Commun 332:43–49PubMedCrossRefGoogle Scholar
  25. 25.
    Shao JY, Gao HY, Li YH, Zhang Y, Lu YY, Zeng YX (2004) Quantitative detection of common deletion of mitochondrial DNA in hepatocellular carcinoma and hepatocellular nodular hyperplasia. World J Gastroenterol 10:1560–1564PubMedGoogle Scholar
  26. 26.
    Shao JY, Li YH, Gao HY, Mai HQ, Zhang Y, Guo X, Zeng YX (2004) High frequency of common deletion (4981 bp) in mitochondrial DNA in nasopharyngeal carcinoma and its correlation with patient age and clinical stages. Cancer Biol Ther 3:1270–1274PubMedCrossRefGoogle Scholar
  27. 27.
    Rogounovitch TI, Saenko VA, Shimizu-Yoshida Y, Abrosimov AY, Lushnikov EF, Roumiantsev PO, Ohtsuru A, Namba H, Tsyb AF, Yamashita S (2002) Large deletions in mitochondrial DNA in radiation-associated human thyroid tumors. Cancer Res 62:7031–7041PubMedGoogle Scholar
  28. 28.
    Lewis PD, Baxter P, Paul GA, Parry JM, Skibinski DO (2000) Detection of damage to the mitochondrial genome in the oncocytic cells of Warthin’s tumour. J Pathol 191:274–281PubMedCrossRefGoogle Scholar
  29. 29.
    Yin PH, Lee HC, Chau GY, Wu YT, Li SH, Lui WY, Wei YH, Liu TY, Chi CW (2004) Alteration of the copy number and deletion of mitochondrial DNA in human hepatocellular carcinoma. Br J Cancer 90:2390–2396PubMedGoogle Scholar
  30. 30.
    Wu CW, Yin PH, Hung WY, Li AF, Li SH, Chi CW, Wei YH, Lee HC (2005) Mitochondrial DNA mutations and mitochondrial DNA depletion in gastric cancer. Genes Chromosomes Cancer 44:19–28PubMedCrossRefGoogle Scholar
  31. 31.
    Lee HC, Yin PH, Yu TN, Chang YD, Hsu WC, Kao SY, Chi CW, Liu TY, Wei YH (2001) Accumulation of mitochondrial DNA deletions in human oral tissues—effects of betel quid chewing and oral cancer. Mutat Res 493:67–74PubMedGoogle Scholar
  32. 32.
    Yang JH, Lee HC, Chung JG, Wei YH (2004) Mitochondrial DNA mutations in light-associated skin tumors. Anticancer Res 24:1753–1758PubMedGoogle Scholar
  33. 33.
    Rossignol R, Gilkerson R, Aggeler R, Yamagata K, Remington SJ, Capaldi RA (2004) Energy substrate modulates mitochondrial structure and oxidative capacity in cancer cells. Cancer Res 64:985–993PubMedCrossRefGoogle Scholar
  34. 34.
    Jia L, Liu KZ, Newland AC, Mantsch HH, Kelsey SM (1999) Pgp-positive leukaemic cells have increased mtDNA but no increased rate of proliferation. Br J Haematol 107:861–869PubMedCrossRefGoogle Scholar
  35. 35.
    Han YC, Kong WJ, Zhang S, Wang YJ, Wang Y, Chen X (2004) Mutation of mitochondrial DNA 4977 bp deletion in laryngeal squamous cell cancer. Ai Zheng 23:1297–1301PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Chuanzhong Ye
    • 1
  • Xiao-Ou Shu
    • 1
  • Wanqing Wen
    • 1
  • Larry Pierce
    • 1
  • Regina Courtney
    • 1
  • Yu-Tang Gao
    • 2
  • Wei Zheng
    • 1
  • Qiuyin Cai
    • 1
  1. 1.Department of MedicineVanderbilt Epidemiology Center, Vanderbilt University School of Medicine and Vanderbilt Ingram-Cancer CenterNashvilleUSA
  2. 2.Department of EpidemiologyShanghai Cancer InstituteShanghaiP.R. China

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