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Expression analysis of activated protein kinase C gene ( LACK 1) in antimony sensitive and resistant Leishmania tropica clinical isolates using real‐time RT‐PCR
Author(s) -
Hajjaran Homa,
KazemiRad Elham,
Mohebali Mehdi,
Oshaghi Mohammad A.,
KhademErfan Mohammad B.,
Hajaliloo Elham,
Reisi Nafchi Hossein,
Raoofian Reza
Publication year - 2016
Publication title -
international journal of dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.677
H-Index - 93
eISSN - 1365-4632
pISSN - 0011-9059
DOI - 10.1111/ijd.13321
Subject(s) - meglumine antimoniate , leishmania tropica , gene , gene expression , real time polymerase chain reaction , biology , reverse transcription polymerase chain reaction , cutaneous leishmaniasis , polymerase chain reaction , complementary dna , drug resistance , leishmania , leishmaniasis , microbiology and biotechnology , genetics , parasite hosting , world wide web , computer science
Background Resistance to pentavalent antimonial drugs has become a serious problem in the treatment of cutaneous leishmaniasis in some endemic areas. Investigations on molecular markers involved in drug resistance are essential for monitoring of the disease. Leishmania ‐activated C kinase gene ( LACK 1) is involved in multiple central processes such as signal transduction. According to the probable role of the LACK 1 gene in antimony resistance, we used real‐time reverse transcription–polymerase chain reaction ( PCR ) to investigate the expression of this gene in clinical L. tropica strains, which were resistant or sensitive to meglumine antimoniate. Methods We analyzed the expression level of LACK in 18 sensitive and 14 resistant L. tropica isolates collected from patients with anthroponotic cutaneous leishmaniasis. After cDNA synthesis, gene expression analysis was performed by quantitative real‐time PCR using SYBR Green. In addition, the full length of the LACK gene from six reference strains was cloned and sequenced then deposited in the NCBI database to confirm our strains. Results Real‐time reverse transcription‐ PCR revealed that the average RNA expression level of LACK in isolates from unresponsive and responsive patients were 0.479 and 4.583, respectively, and expression of LACK was significantly downregulated (9.56‐fold) in resistant isolates compared to sensitive ones. Conclusion Results of the present study suggest the probable role of the LACK gene in antimony resistance. Moreover, it can be considered as a potential marker for monitoring antimony resistance in clinical isolates. However, further studies are required to exploit the biological functions of it in antimony resistance.