Research in Molecular Medicine

en Laurencia caspica extract strongly induces CYP2D6 transcription besides stabilizes its intra-tissue Diazinon-induced transcriptional fluctuations within AGS cells زیست شناسی مولکولی Molecular biology پژوهشي Research Introduction Cytochrome P450 (CYP) isoform 2D6, the second most abundant CYP enzymes in the body involving in the oxidative metabolism of a large number of endogenous and exogenous compounds, is of key relevance for transcription alteration-modulated carcinogenesis. However, absence of a known biological inducer besides of its frequent transcriptional changes made its application challenging. Transactivatory potential of Laurencia caspica extract for CYP2D6 transcription was evaluated in AGS cells in an attempt to rectify its Diazinon-induced fluctuations. Materials and Methods Cytotoxicity of Diazinon and L. caspica ethanol extract as well, was studied on AGS cells using MTT assay. mRNA levels of CYP2D6 were quantitatively estimated using real-time PCR analysis following treatment of the cells with 0-10 μM concentrations of Diazinon or 0-40 μg/ml of L. caspica extract as well as following their co-treatment. The later one, including treatment with 0.1 μM concentration of Diazinon of the cells pre-treated with various concentration ranges of L. capsica extract, was scheduled to evaluate the protective efficacy of L. caspica extract against CYP2D6 transcriptional changes induced following Diazinon treatment. All tests were carried out three times each in triplicate and the results expressed as mean ± standard deviation. Results MTT assay revealed IC50 values of 80 µM and 840 µg/ml for Diazinon and L. capsica extract, respectively. Interestingly, L. capsica extract showed proliferative activity at the concentrations up to 70 µg/ml. qRT-PCR results of the independent experiments from Diazinon treatment revealed irregular transcription fluctuations of CYP2D6. Yet, mean of all data produced a bell-shaped curve in which the 0 and 10 μM treatment caused highest CYP2D6 down-regulation while its transcription remained unchanged with 1 μM Diazinon. When treating with L. caspica extract, CYP2D6 transcription was strongly and steadily incited thereby a 4-fold increase of the mRNA level was induced at highest extract (40 µg/ml) concentration. Similarly, treatment of the cells with 0.1 µM Diazinon of the cells pre-treated with various concentrations of L. caspica extract revealed a dose-dependent transcription up-regulation similar to that of the L. caspica itself, particularly at higher concentrations. Conclusions Ethanol extract from L. caspica has antioxidant and proliferative activities at lower concentrations. It is a strong inducer for CYP2D6 transcription and could rectify Diazinon-induced CYP2D6 fluctuations. Bioactive molecules from L. caspica are promising candidate for detoxification of the chemotherapy drugs particularly in gastric cancer patients.   AGS cell line, CYP2D6, Diazinon, Gastric cancer, Laurencia capsica, Transcription induction 0 0 http://rmm.mazums.ac.ir/browse.php?a_code=A-10-779-3&slc_lang=en&sid=1 Shima Yahoo-Torghabe shimayahoo.t@gmail.com 100319475328460014002 100319475328460014002 No Department of Basic Sciences, Sari Agricultural Sciences and Natural Resources University, Sari, Iran. Zahra Hosseini-Khah zahra_582005@yahoo.com 100319475328460014003 100319475328460014003 No Diabetes Research Center, Mazandaran University of Medical Sciences, Sari, Iran Ali Barzegar alibar647@yahoo.com 100319475328460014004 100319475328460014004 Yes Department of Basic Sciences, Sari Agricultural Sciences and Natural Resources University, Sari, Iran Ali Pakdin-Parizi ali.pakdin@gmail.com 100319475328460014005 100319475328460014005 No Genetics and Agricultural Biotechnology Institute of Tabarestan, Sari Agricultural Sciences and Hossein Dehdari hoseyn_dehdari@yahoo.com 100319475328460014006 100319475328460014006 No Department of Basic Sciences, Sari Agricultural Sciences and Natural Resources University, Sari, Iran Leili Sadeghiamiri leilasadeghi_1370@yahoo.com 100319475328460014007 100319475328460014007 No Tissue Engineering Department, School of Advanced Technologies in Medicine, Mazandaran University of Medical Sciences, Sari, Iran.