G. Zhumina1, G.P. Pogossyan1, K.G. Li2, A.A. Konovalova1,
A.Y. Konkabayeva1*, M.A. Mukasheva1, G.M. Tykezhanova1, N.M. Duzbayeva3, S.S. Shorin1 and A.B. Yeshmagambetova3
1Karagandy State University named after E. A. Buketov, Karagandy, Kazakhstan.
2Laboratory of DNA-diagnostics “Dipner’s health resorts”, Karagandy, Kazakhstan.
3Karaganda Economic University of KazPotrebSoyuz, Karagandy, Kazakhstan.
Purpose. We demonstrated the influence of 1,25D/CA and 1,25D/CUR combinations on Nrf2 expression in primary leukemic cells derived from newly diagnosed CML and AML patients of hematological department of Karaganda Regional Clinical Hospital (Karaganda, Kazakhstan). Methods. The research was conducted by using quantitative real-time PCR. Mononuclear cells were isolated by centrifugation in a Ficoll-PaqueTMPREMIUM density gradient. Cell cultures were incubated in RPMI 1640 medium with test agents. RNA extracted from mononuclear cells was used for cDNA molecules synthesis by reverse transcription. Quantitative real-time PCR was carried out in a DT-322 PCR machine. The relative expression level was quantified by using the 2-ΔΔCtmethod. The statistical analysis was carried out by GraphPadPrism 6.0 program. Results. It was found 1.25D, CUR, CA alone and their combinations do not lead to an increase in the level of expression of the Nrf2 gene in peripheral blood mononuclear cells of AML and CML patients. The expression of NQO1 gene did not change much in the all cell groups of CML patients, but is pronounced in cells of AML patients grown in a medium supplemented with combinations of 1.25D/CUR, 1.25D/CA and CUR, CA alone. 1.25D/CUR and 1.25D/CA combinations do not enhance the expression level of the Nrf2 gene, but increase the amount of its protein, which in turn augments the expression level of the NQO1 gene.
Keywords: Vitamin D (1.25D), curcumin, carnosic acid, Nrf2, NQO1, leukemia patients.