Author/Authors :
Khaleghizadeh, Maryam Department of Microbiology - Damghan Branch Islamic Azad University, Damghan, Iran , Forghanifard, Mohammad Mahdi Department of Biology - Damghan Branch Islamic Azad University, Damghan, Iran , Rad, Abolfazl Cellular and Molecular Research Center - Sabzevar University of Medical Sciences, Sabzevar, Iran , Farshchian, Moein Human Genetic Division - Immunology Research Center - Avicenna Research Institute - Mashhad University of Medical Sciences, Mashhad, Iran , Hejazi, Zahra Department of Microbiology - Damghan Branch Islamic Azad University, Damghan, Iran , Gholamin, Mehran Human Genetic Division - Immunology Research Center - Avicenna Research Institute - Mashhad University of Medical Sciences, Mashhad, Iran , Memar, Bahram Department of Pathology - Imam Reza Hospital - Mashhad University of Medical Sciences, Mashhad, Iran , Abbaszadegan, Mohammad Reza Human Genetic Division - Immunology Research Center - Avicenna Research Institute - Mashhad University of Medical Sciences, Mashhad, Iran
Abstract :
Background: Cancer/Testis Antigens (CTAs) are a subgroup of tumor-associated antigens
which are expressed normally in germ line cells and trophoblast, and aberrantly
in a variety of malignancies. One of the most important CTAs is Developmental Pluripotency
Associated-2(DPPA2) with unknown biological function. Considering the importance
of DPPA2 in developmental events and cancer, preparing a suitable platform
to analyze DPPA2 roles in the cells seems to be necessary.
Methods: In this study, the coding sequence of DPPA2 gene was amplified and cloned
into the retroviral expression vector to produce recombinant retrovirus. The viral particles
were transducted to Esophageal Squamous Cell Carcinoma (ESCC) cell line (KYS
E-30 cells) and the stable transducted cells were confirmed for ectopic expression of
DPPA2 gene by real-time PCR.
Results: According to the critical characteristics of retroviral expression system such as
stable and long time expression of interested gene and also being safe due to deletion
of retroviral pathogenic genes, this system was used to induce expression of DPPA2
gene and a valuable platform to analyze its biological function was prepared. Transduction
results clearly showed efficient overexpression of the gene in target cells in
protein level due to high level of GFP expression.
Conclusion: Such strategies can be used to produce high levels of desired protein in
target cells as a therapeutic target. The produced recombinant cells may present a
valuable platform to analyze the effect of DPPA2 ectopic expression in target cells.
Moreover, the introduction of its potential capacity into the mouse model to evaluate
the tumorigenesis of these cancer cells in vivo leads to an understanding of the biological
importance of DPPA2 in tumorigenesis. In addition, our purified protein can be
used in a mouse model to produce specific antibody developing a reliable detection
of DPPA2 existence in any biological fluid through ELISA system.
Keywords :
Testis , Germ cells , Esophageal squamous cell carcinoma , Carcinogenesis
