Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Mitochondrial DNA Copy Number in the Patients of Korean Polycystic Ovary Syndrome (PCOS)

한국인 다낭성난소증후군 환자에서 미토콘드리아 DNA Copy 수의 정량적 분석

  • Park, Ji-Eun ("Genome Research Center for Infertility and Reproductive Medicine" of Korea Ministry of Health & Welfare) ;
  • Jang, Min-Hee ("Genome Research Center for Infertility and Reproductive Medicine" of Korea Ministry of Health & Welfare) ;
  • Cho, Sung-Won ("Genome Research Center for Infertility and Reproductive Medicine" of Korea Ministry of Health & Welfare) ;
  • Kim, Yoo-Shin (Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University) ;
  • Won, Hyung-Jae (Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University) ;
  • Cho, Jung-Hyun (Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University) ;
  • Baek, Kwang-Hyun (Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University) ;
  • Lee, Sook-Hwan (Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University)
  • 박지은 (보건복지부지정 생식의학 및 불임 유전체 연구센터) ;
  • 장민희 (보건복지부지정 생식의학 및 불임 유전체 연구센터) ;
  • 조성원 (보건복지부지정 생식의학 및 불임 유전체 연구센터) ;
  • 김유신 (포천중문의대 차병원 여성의학연구소 산부인과학교실) ;
  • 원형재 (포천중문의대 차병원 여성의학연구소 산부인과학교실) ;
  • 조정현 (포천중문의대 차병원 여성의학연구소 산부인과학교실) ;
  • 백광현 (포천중문의대 차병원 여성의학연구소 산부인과학교실) ;
  • 이숙환 (포천중문의대 차병원 여성의학연구소 산부인과학교실)
  • Published : 2006.12.31
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Abstract

Objective: We analyzed quantification of mitochondria DNA (mtDNA) to investigate the relationship of mitochondria and pathogenesis of PCOS. Materials and Methods: Peripheral blood samples were collected from 28 patients with PCOS who were under the inclusion criteria for PCOS and from 28 healthy controls. Genomic DNA was used to analyze real-time PCR for mtDNA copy number quantification. The mtDNA copy number was compared between the control and PCOS groups. All data was expressed as mean ${\pm}$ SD. Statistical analysis was assessed by t-test. Results: In this study, the mtDNA $C_T$ was $11.67{\pm}0.422$ in PCOS patients and $11.51{\pm}0.722$ in control group, respectively. The mtDNA copy number was $1726410.71{\pm}407858.591$ the patients of in PCOS and $2167887.51{\pm}252459.28$ in control group (p=0.08), respectively. Conclusion: In our study, using real-time PCR, there was a tendency of lower mtDNA copy number in the patients of PCOS when comparing to the control group even though statistical difference was not significant. However, more extensive analysis is required to clarity relationship between mtDNA copy number and pathogenesis of PCOS.

목 적: 제2형 당뇨의 위험도가 높은 PCOS 환자와 미토콘드리아와의 연관성을 보기 위하여 mitochondria DNA copy 수를 알아보고자 하였다. 연구방법: 연구대상자는 ESHRE의 진단 기준을 만족하는 다낭성난소증후군 여성 28명과 연령이 비슷하며 규칙적인 생리를 하는 여성 28명의 대조군을 대상으로 하였다. 연구대상자들의 genomic DNA는 혈액에서 추출하였으며, 미토콘드리아의 ribosomal RNA 부위를 중합효소 연쇄반응을 통해 증폭한 후 클로닝 하여 표준곡선을 작성한 후, 이를 토대로 다낭성난소증후군 환자의 미토콘드리아 initial quantity를 계산하였다. 결 과: Real-time PCR 결과 다낭성난소증후군 환자의 mtDNA copy 수는 $2,167,887.50{\pm}1,252,459.28$, 정상 대조군은 $1,726,410{\pm}407,858.519$으로 다낭성난소증후군 환자에서 약간 감소하였으나 유의한 차이는 없었다 (p=0.08). 결 론: 본 연구에서는 다낭성난소증후군 환자의 혈액에서 mtDNA copy 수를 조사한 결과, 정상 대조군과 다낭성난소증후군 환자 사이에서 mtDNA copy 수의 유의한 차이가 없었다. 다낭성난소증후군의 병인에는 상당히 복합적인 요소가 있는 것으로 보여지며 그 중 인종적, 지역적 그리고 유전적인 변이가 있는 것으로 보이기 때문에 앞으로 여러 인종에서 더 많은 다낭성난소증후군 환자를 대상으로 연구하여야 될 것으로 사료되는 바이다.

Keywords

References

  1. Carmina E, Lobo RA. Polycystic ovary syndrome (PCOS): arguably the most common endocrinopathy is associated with significant morbidity in women. J Clin Endocrinol Metab 1999; 84: 1897-9 https://doi.org/10.1210/jc.84.6.1897
  2. Futterweit W. Polycystic ovary syndrome : clinical perspectives and management. Obstet Gynecol Surv 1999; 54: 403-13 https://doi.org/10.1097/00006254-199906000-00024
  3. Byun E, Kim HJ, Oh JY, Hong YS, Sung YA. The prevalence of polycystic ovary syndrome in college students from Seoul. J Kor Soc Endocrinol 2005; 20: 120-6 https://doi.org/10.3803/jkes.2005.20.2.120
  4. The Rotterdam ESHREI ASRM-sponsored PCOS consensus workshop group. Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome. Fertil Steril. 2004; 81: 19-25
  5. Ehrmann DA, Bames RB, Rosenfield RL. Polycystic ovary syndrome as a form of functional ovarian hyperandrogenism due to dysregulation of androgen secretion. Endocrinol Rev 1995; 16: 322-53
  6. Legro RS, Kunsehnan AR, Dodson WC, Dunaif A. Prevalence and predictors of risk for type 2 diabetes mellitus and impaired glucose tolerance in polycystic ovary syndrome (a prospective, controlled study in 254 affected women). J Clin Endocrinol Metab 1999; 84: 165-9 https://doi.org/10.1210/jc.84.1.165
  7. Calvo RM, Telleria D, Sancho J, San Millan JL, Escobar-Morreale HF. Insulin gene variable number of tandem repeats regulatory polymorphism is not associated with hyperandrogenism in Spanish women. Fertil Steril 2002; 77: 666-8 https://doi.org/10.1016/S0015-0282(01)03238-1
  8. Ehrmann DA, Tang X, Yoshiuchi I, Cox NJ, Bell GI. Relationship of insulin receptor substrate-l and -2 genotypes to phenotypic features of polycystic ovary syndrome. J Clin Endocrinol Metab 2002; 87: 4297-300 https://doi.org/10.1210/jc.2002-020216
  9. San Millan JL, Corton M, Villuendas G, Sancho J, Peral B, Escobar-Morreale HF. Association of polycystic ovary syndrome with genomic variants related to insulin resistance, type 2 diabetes mellitus, and obesity. J Clin Endocrinol Metab 2004; 89: 2640-6 https://doi.org/10.1210/jc.2003-031252
  10. 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. Sequence and orranization of human mitochondriol genome. Nature 1981; 290(5806): 457 -65 https://doi.org/10.1038/290457a0
  11. Seifer DB, Gardiner AC, Ferreira KA, Peluso JJ. Apoptosis as a function of ovarian reserve in women undergoing in vitro fertilization. Fertil Steril 1996; 66(4): 593-8 https://doi.org/10.1016/S0015-0282(16)58573-2
  12. Trounce I, Byme E, Marzuki S. Decline in skeletal muscle mitochondrial respiratory chain function: possible factor in ageing. Lancet 1989; 25; 1(8639): 637-9
  13. Cavelier L, Johannisson A, Gyllensten U. Analysis of mtDNA copy number and composition of single mitochondrial particles using flow cytometry and PCR. Exp Cell Res 2000; 259: 79-85 https://doi.org/10.1006/excr.2000.4949
  14. Piko L, Taylor KD. Amounts of mitochondrial DNA and abundance of some mitochondrial gene transcripts in early mouse embryos. Dev Biol 1987; 123: 364-74 https://doi.org/10.1016/0012-1606(87)90395-2
  15. Chang SW, Lee SH, Jun HS, Kwack K, Cho SW, Jeong HJ, et al. The quantitative analysis the number of mitochondrial DNA copy using real-time PCR and mitochondrial tRNA mutation analysis at position 3243 in Korean gestational diabetes mellitus. Kor J Obstet Gynecol 2005; 48(4): 978-86
  16. Song J, Oh JY, Sung YA, Pak YK, Park KS, Lee HK. Peripheral blood mitochondrial DNA content is related to insulin sensitivity in offspring of type 2 diabetic patients. Diabetes Care 2001 May; 24(5): 865-9 https://doi.org/10.2337/diacare.24.5.865
  17. Kelley DE, He J, Menshikova EV, Ritov VB. Dysfunction of mitochondria in human skeletal muscle in type 2 diabetes. Diabetes 2002 Oct; 51(10): 2944-50 https://doi.org/10.2337/diabetes.51.10.2944
  18. Readson W, Ross RJM, Sweeney MG, Luxon LM, Pembrey ME, Harding AE, Trembath RC. Diabetes mellitus associsted with a pathogenic point mutation in mitochondrial DNA. Lancet 1992; 340: 1376-9 https://doi.org/10.1016/0140-6736(92)92560-3
  19. Kadowaki H, Kodowaki H, Mori Y, Tobe K, Sakuta R, Suzuki Y, Tanabe Y, Sakura H, Awata T, Goto Y, Hayakawa T, Matsuoka K, Kawamori R, Kamada T, Horai S, Nonaka I, Hagura R, Akanuma Y, Yazaki Y. A subtype of diabetes mellitus associated with a mutation of mitochondrial DNA. New Engl J Med 1994; 330: 962-8 https://doi.org/10.1056/NEJM199404073301403
  20. Yanagisawa K, Uchigata Y, Sanaka M, Sakura H, Minei S, Shimizu M, Kanamuro R, Kadowaki T, Kadowaki T, Omori Y. Mutation in the mitochondrial tRNA leu at position 3243 and spontaneous abortions in Japanese women attending a clinic for diabetic pregnancies. Diabetologia 1995; 38(7): 809 -15 https://doi.org/10.1007/s001250050357
  21. Eung-Ji Lee, Kyong-Jai Yoo, So-Jeong Kim, Sook-Hwan Lee, Kwang Yul Cha, Kwang-Hyun Baek. Single nucleotide polymorphism in exon 17 of the insulin receptor gene is not associated with polycystic ovary syndrome in a Korean population. Frtil Steril 2006; 86(2): 380-4 https://doi.org/10.1016/j.fertnstert.2005.12.073
  22. Diamanti-Kandarakis E, Bartzis MI, Zapanti ED, Spina GC, Filandra FA, Tsianateli TC, Bergiele AT and Kouli CR Polymorphism T${\rightarrow}$C (-34 bp) of gene CYP17 promortor in Greek patients with polycystic ovary syndrome. Fertil Steril 1999; 71: 431-5 https://doi.org/10.1016/S0015-0282(98)00512-3
  23. Marszalek B, Lacinski M, Babych N, Capla E, Biemacka-Lukanty J, Warenik-Szymankiewicz A, et al. Investigations on the genetic polymorphism in the region of CYP17 gene encoding 5'-UTR in patients with polycystic ovarian syndrome. Gynecol Endocrinol 2001; 15: 123-8
  24. Lim SK, Kim MS, Lee SH, Baek KH. Polymorphism of CYP17 and CYP11${\alpha}$ for Polycystic Ovary Syndrome in a Korean Population. Kor J Gene 2002; 24(4): 343-8
  25. Franks S, Gharani N, Waterworth D, et al. The genetic basis of polycystic ovary syndrome. Hum Reprod 1997; 12: 2641-8 https://doi.org/10.1093/humrep/12.12.2641
  26. Gharani N, Waterworth DM, Batty S, et al. Association of the steroid synthesis gene CYP11${\alpha}$ with polycystic ovary syndrome and hyperandrogenism. Hum Mol Genet 1997; 6(3): 397-402 https://doi.org/10.1093/hmg/6.3.397
  27. Kandarakis ED, Bartzis MI, Bergiele AT, et al. Microsatellite polymorphism (tttta)n at 528 base pairs of gene CYP11${\alpha}$ influences hyperandrogenemia in parients with polycystic ovary symdrome. Fertil Steril 2000; 73: 735-41 https://doi.org/10.1016/S0015-0282(99)00628-7
  28. Moore CCD, Brentano ST, Miller WL. Human P450sec gene transcription is induced by cyclic AMP and repressed by 12-O-tetradecanoylphorbol-13-acetate and A23817 through independent cis elements. Mol Cell Biol 1990; 10: 6013-23 https://doi.org/10.1128/MCB.10.11.6013