DEVELOPMENT OF AMPLIFICATION REFRACTORY MUTATION SYSTEM PCR TO DETECT ANDROGEN RECEPTOR GENE G1773A (RS6152) POLYMORPHISM
Keywords:
Androgen receptor, rs6152, ARMS-PCR, polymorphism
Abstract
Androgen receptor (AR) is a ubiquitous receptor responsible for responses by androgen stimulus. Androgen, a hormone which will bind to the AR, is essentials for normal male sexual development. Nevertheless, one of the polymorphisms in the AR gene, G1733A (rs6152) have been associated with numerous clinical risks such as cardiovascular disease (CD), androgenetic alopecia, high prostate-specific antigen (PSA) levels, male infertility, recurrent spontaneous abortions and prostate cancer. This study aims to develop an alternative and cost-effective method to detect G1773A (rs6152) polymorphism. In this study, amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) using two pair of primers was used to detect the G1733A (rs6152) polymorphism. Primer design was done using http://primer1.soton.ac.uk/primer1.html online tools and then manually adjusted to increase the specificity. A total of 54 samples were screened using ARMS PCR and 2 representative samples of each allele from previous screening were used to validate the results using Sanger DNA sequencing. Among 54 subjects screened, we found 52 (96.3%) subjects carry G allele and 2 (3.7%) subjects carry A allele. No heterozygote was found in this study. The frequency of G allele was 96.97% and the frequency of A allele was 3.03%. Result validation using DNA sequencing was in agreement with ARMS-PCR method results. ARMS-PCR can be used as efficient alternative for genotyping of G1733A (rs6152) AR gene polymorphism
Downloads
Download data is not yet available.
References
1. Murashima A, Kishigami S, Thomson A, Yamada G. Androgens and mammalian male reproductive tract development. Biochim Biophys Acta - Gene Regul Mech. 2015 Feb 1;1849(2):163–70.
2. Notelovitz M. Androgen effects on bone and muscle. Fertil Steril [Internet]. 2002 [cited 2022 Nov 28];77(SUPPL. 4):34–41. Available from: https://pubmed.ncbi.nlm.nih.gov/12007900/
3. Wilson S, Qi J, Filipp F V. Refinement of the androgen response element based on ChIP-Seq in androgen-insensitive and androgen-responsive prostate cancer cell lines. Sci Rep [Internet]. 2016 Sep 14 [cited 2022 Nov 28];6. Available from: https://pubmed.ncbi.nlm.nih.gov/27623747/
4. Gulía C, Baldassarra S, Zangari A, Briganti V, Gigli S, Gaffi M. Androgen insensitivity syndrome.
5. Kucerova R, Bienova M, Kral M, Bouchal J, Trtkova KS, Burdova A, et al. Androgenetic alopecia and polymorphism of the androgen receptor gene (SNP rs6152) in patients with benign prostate hyperplasia or prostate cancer. J Eur Acad Dermatol Venereol [Internet]. 2015 Jan 1 [cited 2022 Oct 13];29(1):91–6. Available from: https://pubmed.ncbi.nlm.nih.gov/24665929/
6. Porras-Dorantes Á, Brambila-Tapia AJL, Lazcano-Castellanos AB, Da Silva-José TD, Juárez-Osuna JA, García-Ortiz JE. Association between G1733A (rs6152) polymorphism in androgen receptor gene and recurrent spontaneous abortions in Mexican population. J Assist Reprod Genet [Internet]. 2017 Oct 1 [cited 2022 Feb 26];34(10):1303. Available from: /pmc/articles/PMC5633579/
7. Peng CY, Long XY, Lu GX. Association of AR rs6152G/A gene polymorphism with susceptibility to polycystic ovary syndrome in Chinese women. Reprod Fertil Dev [Internet]. 2010 May 20 [cited 2022 Nov 29];22(5):881–5. Available from: https://www.publish.csiro.au/rd/RD09190
8. Agiannitopoulos K, Bakalgianni A, Marouli E, Zormpa I, Manginas A, Papamenzelopoulos S, et al. Gender Specificity of a Genetic Variant of Androgen Receptor and Risk of Coronary Artery Disease. J Clin Lab Anal [Internet]. 2016 May 1 [cited 2022 Feb 26];30(3):204. Available from: /pmc/articles/PMC6807026/
9. Robles-Fernandez I, Martinez-Gonzalez LJ, Pascual-Geler M, Cozar JM, Puche-Sanz I, Serrano MJ, et al. Association between polymorphisms in sex hormones synthesis and metabolism and prostate cancer aggressiveness. PLoS One [Internet]. 2017 Oct 1 [cited 2022 Nov 29];12(10):e0185447. Available from: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0185447
10. Rana K, Davey RA, Zajac JD. Human androgen deficiency: insights gained from androgen receptor knockout mouse models. Asian J Androl [Internet]. 2014 Mar [cited 2022 Feb 26];16(2):169–77. Available from: https://pubmed.ncbi.nlm.nih.gov/24480924/
11. Morova T, McNeill DR, Lallous N, Gönen M, Dalal K, Wilson DM, et al. Androgen receptor-binding sites are highly mutated in prostate cancer. Nat Commun [Internet]. 2020 Dec 1 [cited 2022 Dec 20];11(1). Available from: /pmc/articles/PMC7012874/
12. Henningsson S, Jonsson L, Ljunggren E, Westberg L, Gillberg C, Råstam M, et al. Possible association between the androgen receptor gene and autism spectrum disorder. Psychoneuroendocrinology [Internet]. 2009 Jun [cited 2022 Dec 20];34(5):752–61. Available from: https://pubmed.ncbi.nlm.nih.gov/19167832/
13. Dewi AL, Paramita DK, Facliiroli J. Development of Tetra-primer Amplification Refractory Mutation System (ARMS) PCR for Detection of CHRNA3 rs8040868. Indones Biomed J [Internet]. 2021 Jun 14 [cited 2022 Dec 20];13(2):192–200. Available from: https://inabj.org/index.php/ibj/article/view/1463
14. Alyethodi RR, Singh U, Kumar S, Alex R, Deb R, Sengar GS, et al. T-ARMS PCR genotyping of SNP rs445709131 using thermostable strand displacement polymerase. BMC Res Notes [Internet]. 2018 Feb 15 [cited 2022 Dec 20];11(1):1–5. Available from: https://bmcresnotes.biomedcentral.com/articles/10.1186/s13104-018-3236-6
15. Mehdi M, 1 H, Hadadzadeh M, Fallahzadeh H. Development of One-Step Tetra-primer ARMS-PCR for Simultaneous Detection of the Angiotensin Converting Enzyme (ACE) I/D and rs4343 Gene Polymorphisms and the Correlation with CAD Patients. Avicenna J Med Biotechnol [Internet]. 2019 [cited 2022 Dec 20];11(1):118. Available from: /pmc/articles/PMC6359703/
16. Jin C, Li Z, Zheng X, Shen K, Chao J, Dong Y, et al. Development and validation of T‐ARMS‐PCR to detect CYP2C19*17 allele. J Clin Lab Anal [Internet]. 2020 Jan 1 [cited 2022 Dec 20];34(1). Available from: /pmc/articles/PMC6977150/
17. Yang L, Ijaz I, Cheng J, Wei C, Tan X, Khan MA, et al. Evaluation of amplification refractory mutation system (ARMS) technique for quick and accurate prenatal gene diagnosis of CHM variant in choroideremia. Appl Clin Genet [Internet]. 2017 Dec 19 [cited 2023 Jan 16];11:1–8. Available from: https://www.dovepress.com/evaluation-of-amplification-refractory-mutation-system-arms-technique--peer-reviewed-fulltext-article-TACG
18. Garbuz MM, Ovchinnikova AA, Kumeiko VV. Design, Optimization and Validation of the ARMS PCR Protocol for the Rapid Diagnosis of Wilson’s Disease Using a Panel of 14 Common Mutations for the European Population. Genes 2022, Vol 13, Page 1940 [Internet]. 2022 Oct 25 [cited 2023 Jan 16];13(11):1940. Available from: https://www.mdpi.com/2073-4425/13/11/1940/htm
19. Medrano RFV, De Oliveira CA. Guidelines for the Tetra-Primer ARMS–PCR Technique Development. Mol Biotechnol 2014 567 [Internet]. 2014 Feb 12 [cited 2022 Dec 20];56(7):599–608. Available from: https://link.springer.com/article/10.1007/s12033-014-9734-4
20. Sribudiani Y, Marwan DW, Aulanni’am A, Widodo MA, Purnomo BB, Panigoro R, et al. Germline Mutations and Polymorphisms of Androgen Receptor in Prostate Cancer Patients: Frequency and Results of in Silico Analysis. Asian Pac J Cancer Prev [Internet]. 2018 Aug 1 [cited 2022 Dec 20];19(8):2241. Available from: /pmc/articles/PMC6171394/
2. Notelovitz M. Androgen effects on bone and muscle. Fertil Steril [Internet]. 2002 [cited 2022 Nov 28];77(SUPPL. 4):34–41. Available from: https://pubmed.ncbi.nlm.nih.gov/12007900/
3. Wilson S, Qi J, Filipp F V. Refinement of the androgen response element based on ChIP-Seq in androgen-insensitive and androgen-responsive prostate cancer cell lines. Sci Rep [Internet]. 2016 Sep 14 [cited 2022 Nov 28];6. Available from: https://pubmed.ncbi.nlm.nih.gov/27623747/
4. Gulía C, Baldassarra S, Zangari A, Briganti V, Gigli S, Gaffi M. Androgen insensitivity syndrome.
5. Kucerova R, Bienova M, Kral M, Bouchal J, Trtkova KS, Burdova A, et al. Androgenetic alopecia and polymorphism of the androgen receptor gene (SNP rs6152) in patients with benign prostate hyperplasia or prostate cancer. J Eur Acad Dermatol Venereol [Internet]. 2015 Jan 1 [cited 2022 Oct 13];29(1):91–6. Available from: https://pubmed.ncbi.nlm.nih.gov/24665929/
6. Porras-Dorantes Á, Brambila-Tapia AJL, Lazcano-Castellanos AB, Da Silva-José TD, Juárez-Osuna JA, García-Ortiz JE. Association between G1733A (rs6152) polymorphism in androgen receptor gene and recurrent spontaneous abortions in Mexican population. J Assist Reprod Genet [Internet]. 2017 Oct 1 [cited 2022 Feb 26];34(10):1303. Available from: /pmc/articles/PMC5633579/
7. Peng CY, Long XY, Lu GX. Association of AR rs6152G/A gene polymorphism with susceptibility to polycystic ovary syndrome in Chinese women. Reprod Fertil Dev [Internet]. 2010 May 20 [cited 2022 Nov 29];22(5):881–5. Available from: https://www.publish.csiro.au/rd/RD09190
8. Agiannitopoulos K, Bakalgianni A, Marouli E, Zormpa I, Manginas A, Papamenzelopoulos S, et al. Gender Specificity of a Genetic Variant of Androgen Receptor and Risk of Coronary Artery Disease. J Clin Lab Anal [Internet]. 2016 May 1 [cited 2022 Feb 26];30(3):204. Available from: /pmc/articles/PMC6807026/
9. Robles-Fernandez I, Martinez-Gonzalez LJ, Pascual-Geler M, Cozar JM, Puche-Sanz I, Serrano MJ, et al. Association between polymorphisms in sex hormones synthesis and metabolism and prostate cancer aggressiveness. PLoS One [Internet]. 2017 Oct 1 [cited 2022 Nov 29];12(10):e0185447. Available from: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0185447
10. Rana K, Davey RA, Zajac JD. Human androgen deficiency: insights gained from androgen receptor knockout mouse models. Asian J Androl [Internet]. 2014 Mar [cited 2022 Feb 26];16(2):169–77. Available from: https://pubmed.ncbi.nlm.nih.gov/24480924/
11. Morova T, McNeill DR, Lallous N, Gönen M, Dalal K, Wilson DM, et al. Androgen receptor-binding sites are highly mutated in prostate cancer. Nat Commun [Internet]. 2020 Dec 1 [cited 2022 Dec 20];11(1). Available from: /pmc/articles/PMC7012874/
12. Henningsson S, Jonsson L, Ljunggren E, Westberg L, Gillberg C, Råstam M, et al. Possible association between the androgen receptor gene and autism spectrum disorder. Psychoneuroendocrinology [Internet]. 2009 Jun [cited 2022 Dec 20];34(5):752–61. Available from: https://pubmed.ncbi.nlm.nih.gov/19167832/
13. Dewi AL, Paramita DK, Facliiroli J. Development of Tetra-primer Amplification Refractory Mutation System (ARMS) PCR for Detection of CHRNA3 rs8040868. Indones Biomed J [Internet]. 2021 Jun 14 [cited 2022 Dec 20];13(2):192–200. Available from: https://inabj.org/index.php/ibj/article/view/1463
14. Alyethodi RR, Singh U, Kumar S, Alex R, Deb R, Sengar GS, et al. T-ARMS PCR genotyping of SNP rs445709131 using thermostable strand displacement polymerase. BMC Res Notes [Internet]. 2018 Feb 15 [cited 2022 Dec 20];11(1):1–5. Available from: https://bmcresnotes.biomedcentral.com/articles/10.1186/s13104-018-3236-6
15. Mehdi M, 1 H, Hadadzadeh M, Fallahzadeh H. Development of One-Step Tetra-primer ARMS-PCR for Simultaneous Detection of the Angiotensin Converting Enzyme (ACE) I/D and rs4343 Gene Polymorphisms and the Correlation with CAD Patients. Avicenna J Med Biotechnol [Internet]. 2019 [cited 2022 Dec 20];11(1):118. Available from: /pmc/articles/PMC6359703/
16. Jin C, Li Z, Zheng X, Shen K, Chao J, Dong Y, et al. Development and validation of T‐ARMS‐PCR to detect CYP2C19*17 allele. J Clin Lab Anal [Internet]. 2020 Jan 1 [cited 2022 Dec 20];34(1). Available from: /pmc/articles/PMC6977150/
17. Yang L, Ijaz I, Cheng J, Wei C, Tan X, Khan MA, et al. Evaluation of amplification refractory mutation system (ARMS) technique for quick and accurate prenatal gene diagnosis of CHM variant in choroideremia. Appl Clin Genet [Internet]. 2017 Dec 19 [cited 2023 Jan 16];11:1–8. Available from: https://www.dovepress.com/evaluation-of-amplification-refractory-mutation-system-arms-technique--peer-reviewed-fulltext-article-TACG
18. Garbuz MM, Ovchinnikova AA, Kumeiko VV. Design, Optimization and Validation of the ARMS PCR Protocol for the Rapid Diagnosis of Wilson’s Disease Using a Panel of 14 Common Mutations for the European Population. Genes 2022, Vol 13, Page 1940 [Internet]. 2022 Oct 25 [cited 2023 Jan 16];13(11):1940. Available from: https://www.mdpi.com/2073-4425/13/11/1940/htm
19. Medrano RFV, De Oliveira CA. Guidelines for the Tetra-Primer ARMS–PCR Technique Development. Mol Biotechnol 2014 567 [Internet]. 2014 Feb 12 [cited 2022 Dec 20];56(7):599–608. Available from: https://link.springer.com/article/10.1007/s12033-014-9734-4
20. Sribudiani Y, Marwan DW, Aulanni’am A, Widodo MA, Purnomo BB, Panigoro R, et al. Germline Mutations and Polymorphisms of Androgen Receptor in Prostate Cancer Patients: Frequency and Results of in Silico Analysis. Asian Pac J Cancer Prev [Internet]. 2018 Aug 1 [cited 2022 Dec 20];19(8):2241. Available from: /pmc/articles/PMC6171394/
Published
2023-07-29
How to Cite
Triyana Sari, Erick Sidarta, Sari Mariyati Dewi Nataprawira, Giovani Evangelista Alfonsius, & Damar Sajiwo. (2023). DEVELOPMENT OF AMPLIFICATION REFRACTORY MUTATION SYSTEM PCR TO DETECT ANDROGEN RECEPTOR GENE G1773A (RS6152) POLYMORPHISM . Jurnal Scientia, 12(03), 1-7. Retrieved from https://infor.seaninstitute.org/index.php/pendidikan/article/view/2952
Section
Articles
