Measurement of Human Progesterone Receptor A Gene Expression in Normal and Breast Cancer Tissue using Real-Time PCR Technique

Document Type : Original Article


1 M.Sc. Student of Genetics, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.

2 Associate Professor Cancer Genetics, Biotechnology Research Institute, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.

3 Assistant Professor of Genetics and Breeding, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.

4 Fellowship of Gastrointestinal Pathology, Razavi Hospital Vice President for Research Cancer, Mashhad, Iran.

5 Professor of Veterinary Pathology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.

6 PhD. Student of Genetics, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.


Introduction: The improvement of knowledge in the field of molecular genetics and diagnostics of breast cancer can help us better to understand the biological process that leads a normal breast cell to be transformed into a malignant one with invasive and metastatic confidants. The aim of this study was to introduce the Relative Quantitative Real-time PCR technique based on sybr green dye for relative quantification of progesterone receptor A (PR-A) gene expression in normal and malignant neoplasia in human breast tissue.
Methods: 20 malignant samples (Grad 2) and 10 normal samples of human breast tissue were collected before treatment and without contamination. After RNA extraction and cDNA synthesis, PR-A and GAPDH genes for measurement of relative quantification were amplified by Real-time PCR according to the standard method. The obtained Ct value were analyzed by SDS version 1.4 software, GLM program and T-test student procedure were performed in SAS version 9.1 software to determine statistical significance.
Results: PR-A gene can be expressed as in both normal and malignant breast tissue but expression of PR-A gene in malignant breast tissue was over expressed than normal cases (p≤0/05). The statistical significance difference test for comparing of PR-A gene expression in malignant breast tissue to the mean of normal samples showed an increased in 70 percent of samples. Also there was no significant correlation between over expression of PR-A gene and marital status and menopause (p ≤0.05). Furthermore, results indicated a linear correlation was between expression of PR-A gene and the age of cancer patients (p ≤0.05).
Conclusion: PR-A gene can be used as a biomarker for prognosis, screening and diagnosis of malignant patients against normal persons at the beginning stages of breast cancer.


  1. Mousavi SM, Montazeri A, Mohagheghi MA, Jarrahi AM, Harirchi I, Najafi M, et al Breast cancer in Iran: an epidemiological review. Breast J 2007 Jul-Aug;13(4):383-91.
  2. Harirchi I, Ebrahimi M, Zamani N, Jarvandi S, Montazeri A. Breast cancer in Iran. a review of 903 case records. Public Health 2000 Mar;114(2):143-5.
  3. Tabor MP, Brakenhoff RH, Ruijter-Schippers HJ, Kummer JA, Leemans CR, Braakhuis BJ. Genetically altered fields as origin of locally recurrent head and neck cancer: a retrospective study. Clin Cancer Res 2004 Jun 1; 10(11):3607-13.
  4. Behjati F, Atri M, Najmabadi H, Nouri K, Zamani M, Mehdipour P. Prognostic value of chromosome 1 and 8 copy number in invasive ductal breast carcinoma among Iranian women: an interphase FISH analysis. Pathol Oncol Res 2005 Sep;11(3):157-63.
  5. Yassaee VR, Zeinali S, Harirchi I, Jarvandi S, Mohagheghi MA, Hornby DP, et al. Novel mutation in the BRCA1 and BRCA2 genes in Iranian women with early-onset breast cancer. Breast Cancer Res 2002;4(4):R6.
  6. Lin Y, Kikuchi S, Tamakoshi K, Kondo T, Niwa Y, Yatsuya H, et al. Active smoking, passive smoking and breast cancer risk: findings from the Japan Collaborative Cohort Study for Evaluation of Cancer Risk. J Epidemiol 2008;18(2):77-83.
  7. Abba MC, Hu Y, Sun H, Drake JA, Gaddis S, Baggerly K, et al. Gene expression signature of estrogen receptor α. status in breast cancer. BMC Genomics 2005 Mar;37(6):1-13.
  8. Christensen SA. Receptors in breast cancer: interpreting estrogen, progesterone, and HER-2 status. Available at: (2009 Sep).
  9. Camacho-Arroyo I, Rodríguez-Dorantes M. Transcriptional activity regulated by progesterone receptor isoforms. Mol Endocrinol 2006;25-38.
  10. Vandesompele J, De Paepe A, Speleman F. Elimination of primer-dimer artifacts and genomic coamplification using a two-step SYBR green I real-time RT-PCR. Anal Biochem 2002 Apr;303(1):95-8.
  11. Pfaffle MW. A new mathematical model for relative quantification in realtime RT-PCR. Nucleic Acids Res 2001 May;29(9):e45.
  12. Graham JD, Clark CL. Expression and transcriptional activity of progesterone receptors A and progesterone receptor B in mammalian cells. Breast Cancer Res 2002 Jul;4(5):187–90.
  13. Graham JD, Yeates C, Balleine RL, Harvey SS, Milliken JS, Bilous AM, et al. Characterization of progesterone receptor A and B expression in human breast cancer. Cancer Res 1995 Nov 1;55(21):5063-8.
  14. Hopp TA, Weiss HL, Hilsenbeck SJ, Cui Y, Allred DC, Horwitz KB, et al. Breast cancer patients with progesterone receptor PR-A-rich tumors have poorer disease-free survival rates. Clin Cancer Res 2004 Apr 15;10(8):2751-60.
  15. Graham JD, Mote PA, Salagame U, van Dijk JH, Balleine RL, Huschtscha LI, et al. DNA replication licensing and progenitor numbers are increased by progesterone in normal human breast. Endocrinology 2009 Jul;150(7):3318-26.
  16. Lydon JP, Edwards EP. Finally! A model for progesterone receptor action in normal human breast. Endocrinology 2009 Jul;150(7):2988-90.
  17. Mote PA, Bartow S, Tran N, Clarke CL. Loss of co-ordinate expression of progesterone receptors A and B is an early event in breast carcinogenesis. Breast Cancer Res 2002 Mar;72(2):163-72.
  18. Hasan TN, B LG, Masoodi TA, Shafi G, Alshatw AA, Sivashanmugham P. Affinity of estrogens for human progesterone receptor A and B monomers and risk of breast cancer: a comparative molecular modeling study. Adv Appl Bioinform Chem 2011 Mar;4:29-36.
  19. Jacobsen BM, Schittone SA, Richer JK, Horwitz KB. Progesterone-independent effects of human progesterone receptors (PRs) in estrogen receptor-positive breast cancer: PR isoform-specific gene regulation and tumour biology. Mol Endocrinol 2005 Mar;19(3): 574-87.