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Abstract(Please copy/paste the abstract send to the congress) : |
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Protozoan parasites
Expression analysis of Activated protein Kinase C gene (LACK1) in antimony sensitive and resistant Leishmania tropica clinical isolates using Real-Time RT-PCR
Hajjaran H.1, Kazemi-Rad E. 2, Mohebali M. 1, Oshaghi M.A. 3, Khadem Erfan M.B. 4, Hajialilo E. 1, Reissi H. 1, Raoofian R. 5
1Tehran University of Medical Sciences, Dep. Medical Parasitology and Mycology, Tehran
2Department of Parasitology, Pasteur Institute of Iran, Tehran
3Tehran University of Medical Sciences,, Departments of Medical Entomology and Vector Control , Tehran
4Kurdistan University of Medical Sciences, , Department of Medical parasitology and Mycology,, Sanandaj
5Legal Medicine Research Center, Legal Medicine Organization, Iran, Tehran,
Background: Resistance to pentavalent antimonial drugs has become a serious problem in treatment of cutaneous leishmaniasis (CL) in some endemic areas. Investigations on molecular markers involved in drug resistance are essential for monitoring of the disease. Leishmania activated C kinase gene (LACK1) is involved in multiple central processes including signal transduction, RNA processing, and cell cycle control. According to the probable role of the LACK gene in antimony resistance, we used real-time RT-PCR to investigate the expression of this gene in clinical L. tropica strains which were resistant or sensitive to meglumine antimoniate (Glucantimee).
Methods: we analyzed the relative expression level of LACK in 18 sensitive and 14 resistant Leishmania tropica clinical isolates, were collected from anthroponotic cutaneous leishmaniasis (ACL) patients. After cDNA synthesis, gene expression analysis was performed by quantitative real-time PCR using SYBR® Green. In addition, full length of LACK gene from 6 reference strains was cloned and sequenced then deposited in NCBI database to confirm our strains.
Results: Real time RT-PCR revealed that the average of the RNA expression level of LACK gene in isolates from unresponsive and responsive patients were 0.479 and 4.583 respectively and expression of LACK was significantly down regulated (9.56 fold) in resistant isolates compared to sensitive ones.
Conclusion: The results of the present study suggest the probable role of LACK gene in antimony resistance. Moreover, it can be considered as a potential marker for monitoring of antimony resistance in clinical isolates. However, further studies are required to exploit the biological functions of it in antimony resistance.
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