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Pitfall of Real-Time PCR Method to Find Delta mtDNA in Colon Cancer

Author Affiliations

  • 1Ghaffarpour Massoud, Hasrak Kambiz, and Houshmand Massoud Department of Molecular Medicine and Genetics, School of Medicine, Hamedan University of Medical Sciences, Hamedan, IRAN
  • 2 National Institute for Genetic Engineering and Biotechnology (NIGEB), Tehran, IRAN
  • 3 Medical Genetics Department, Special Medical Center, Tehran, IRAN

Int. Res. J. Biological Sci., Volume 2, Issue (11), Pages 55-59, November,10 (2013)

Abstract

In this study, we investigated the presence of a high incidence of common mitochondrial deletion in mtDNA of colon cancer tissues and in blood of both patients and control samples. For this purpose, one of the common regions of the mitochondrial genome was screened using different techniques including PCR amplification, Southern blot, and real-time PCR followed by DNA sequence analysis. Isolated DNA was applied for amplification of hypervariable regions ATPase8/6, COXIII, ND3, ND4, and ND5 of delta mtDNA4977. In 25 colon cancer patients, delta mtDNA4977 was identified using quantitative RT-PCR in 6 cases (4.5%) of the tumoral tissues and 18 cases (4.47%) of the non-tumoral tissues that were adjacent to the tumors. Delta mtDNA4977 was detected more in non-tumoral tissues than in adjacent tumoral tissues, but less common deletion was observed (0.017%). Additionally, no 4,977-bp common mitochondrial deletion was observed in blood of patients and 100 control samples. Unknown genetic aspects, ambiguous environmental factors, and reactive oxygen species (ROS) are events that cause the delta mtDNA4977 mutation rate to be elevated more in non-tumoral tissues as compared to adjacent tumoral tissues of colon cancer. The results of our study propose that delta mtDNA4977 percentage in tumoral tissue is less prevalent and is untolerable, perhaps due to high metabolism and ROS production.

References

  1. Salimzadeh H., Delavari A., Montazeri A. and Mirzazadeh A., Knowledge and Practice of Iranians Toward Colorectal Cancer, and Barriers to Screening, Int. J. Prev. Med., 3(1), 29-35 (2012)
  2. Davari M., Maracy M.R., Emami M.H., Taheri D., Aslani A., Givi M. and Massah S, The Direct Medical Ccosts of Colorectal Cancer in Iran; Analyzing the Patient’s Level Data from a Cancer Specific Hospital in Isfahan, Int. J. Prev. Med., 3(12), 887-892 (2012)
  3. Sarmandal CV., Cancer, Heart and other Chronic Diseases: Some Preventive Measures to Control Lipid Peroxidation through Choice of Edible Oils, International Research Journal of Biological Sciences, 1(6), 68-75 (2012)
  4. Iqbal S., Raffat S.K., Siddiqui M.S. and Siddiq M.,Social Influence of Biological Viruses on Communities, International Research Journal of Biological Sciences, 1(8), 71-75 (2012)
  5. Sirappuselvi S., and Chitra M., In vitro Antioxidant Activity of Cassia tora Lin, International Research Journal of Biological Sciences,1(6), 57-61 (2012)
  6. Aweng E.R., Hanisah N., Mohd Nawi M.A.,, Nurhanan Murni Y., Shamsul M., Antioxidant Activity and Phenolic Compounds of Vitex Trifolia Var, Simplicifolia Associated with Anticancer, International Research Journal of Biological Sciences, 1(3), 65-68, (2012)
  7. Lowe H.I., Watson C.T., Badal S., Ateh E.N., Toyang N.J., and Bryant J., Anti-angiogenic properties of the Jamaican ball moss, (Tillandsia recurvata L.), International Research Journal of Biological Sciences, 1(4), 73-76 (2012)
  8. Zhou S., Kassauei K., Cutler D.J., Kennedy G.C., Sidransky D., Maitra A. and Califano J., An Oligonucleotide Microarray for High-Throughput Sequencing of the Mitochondrial Genome, J. Mol. Diagn.,8(4), 476-482 (2006)
  9. Robin E.D. and Wong R., Mitochondrial DNA Molecules and Virtual Number of Mitochondria Per Cell in Mammalian Cells, J. Cell. Physiol., 136(3), 507-513 (1988)
  10. Brown W.M., George M. and Wilson A.C., Rapid Evolution of Aanimal Mitochondrial DNA, Proc. Natl. Acad. Sci., 76(4), 1967-1971 (1979)
  11. Li Y., Zhou H., Stansbury K. and Trush M., Role of reactive oxygen species in multistage carcinogenesis. In: Thomas C. and Kalyanaram B. editors. Oxygen radicals and disease process, 1977. Amsterdam: Harwood Academic Publishers, 237-277 (1997)
  12. Kroemer G. and Reed J.C., Mitochondrial control of cell death, Nat. Med., 6(5), 513-519 (2000)
  13. Dani M.A., Dani S.U., Lima S.P., Martinez A., Rossi B.M., Soares F., Zago M.A. and Simpson, A.J., Less DeltamtDNA4977 than Normal in Various Types of Tumors Suggests that Cancer Cells Are Essentially Free of This Mutation, Genet. Mol. Res., 3(3), 395-409 (2004)
  14. Wei Y.H., Mitochondrial DNA Alterations as Ageing-Associated Molecular Events, Mutat. Res., 275(3-6), 145-155 (1992)
  15. Bandy B. and Davison A.J., Mitochondrial Mutations May Increase Oxidative Stress: Implications for Carcinogenesis and Aging?, Free Radic. Biol. Med., 8(6), 523-539 (1990)
  16. Bohr V., Anson R.M., Mazur S. and Dianov G., Oxidative DNA Damage Processing and Changes with Aging, Toxicol. Lett., 28(102-103), 47-52 (1998)
  17. Rogounovitch T.I., Saenko V.A., Shimizu-Yoshida Y., Abrosimov A.Y., Lushnikov EF., Roumiantsev P.O., Ohtsuru A., Namba H., Tysb A.F. and Yamashita S. Large Deletions in Mitochondrial DNA in Radiation-Associated Human Thyroid Tumors, Cancer Res., 62(23), 7031-7041(2002)
  18. Kawashima S., Ohta S., Kagawa Y., Yoshida M. and Nishizawa M., Widespread Tissue Distribution of Multiple Mitochondrial DNA Deletions in Familial Mitochondrial Myopathy. Muscle Nerve., 17(7), 741-746 (1994)
  19. Lee H.C., Pang C.Y., Hsu H.S. and Wei Y.H., Differential Accumulations of 4,977 bp Deletion in Mitochondrial DNA of Various Tissues in Human Ageing, Biochim Biophys Acta., 226(1), 37-43 (1994)
  20. Sandip S.M., A Reliable and High Yielding Method for Isolation of Genomic DNA from Ammi majus, International Research Journal of Biological Sciences, 2(1), 57-60, (2013)
  21. Tan D.J., Chang J., Chen W.L., Agress L.I, Yeh K.T., Wang B. and Wong L.I., Novel Heteroplasmic Frameshift and Missense Somatic Mitochondrial DNA Mutations in Oral Cancer of Betel Quid Chewers, Genes Chromosomes Cancer., 37(2), 186-194 (2003)
  22. Turker M.S. Somatic Cell Mutations: Can They Provide a Link between Aging and Cancer?, Mech. Agei Dev., 117(1-3), 1-19 (2000)
  23. Turner C., Killoran C., Thomas N.S., Rosenberg M., Chuzhanova N.A., Johnston J., Kemel Y., Cooper D.N. and Biesecker L.G., Human Genetic Disease Caused by de novo Mitochondrial-Nuclear DNA Transfer, Hum. Genet., 112(3), 303-309 (2003)
  24. Warowicka A., Kwasniewska A. and Gozdzicka-Jozefiak A., Alterations in mtDNA: A Qualitative and Quantitative Study Associated with Cervical Cancer Development, Gyneco. Oncol., 129(1), 193-198 (2013)
  25. Zheng Y., Luo X., Zhu J., Zhang X., Cheng H., Xiz Z., Su N., Zhang N. and Zhou J, Mitochondrial DNA 4977 bp Deletion Is a Common Pphenomenon in Hair and Increases with Age. Bosn. J. Basic Med. Sci., 12(3), 187-192 (2012)
  26. Chen T., He J., Shen L., Fang H., Nie H., Jin T., Wei X., Xin Y., Jiang Y., Li H., Chen G., Lu J. and Bai Y., The Mitochondrial DNA 4,977-bp Deletion and Its Implication in Copy Number Alteration in Colorectal Cancer, BMC Med. Genet., 13(12), 8 (2011)
  27. Kamalidehghan B. and Houshmand M., Pitfalls for Common Mitochondrial DNA Deletion (mtDNA4977) As a Biomarker of Cancer, Arch. Med. Res., 44(1), 79-80 (2013)