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Analysis of Akt protein expression in CaCo2 and NCI-H358 cells treated with N-sulfonylpurine derivatives by Western blotting

Most of anticancer drugs are not target-specific, affecting normal cells also. Newly synthesized N-9-sulfonylpurine derivatives belong to the group of antimetabolites that achieve specific effects through interaction with natural metabolites. The aim of this study is to determine possible changes in the expression of the Akt protein, a central effector in the PI3K signalling pathway, in adenocarcinoma (CaCo-2) and bronchioalveolar carcinoma (NCI-H358) cells after treatment with two N-9-sufonylpurine derivatives. After 24 hrs of treatment cells were lysed and cellular proteins were isolated. Protein concentration was determined by the BCA protein assay with bovine serum albumin as a standard. Presence of Akt protein in total isolated cellular proteins was determined by the Western blot method using Akt antibody. Relative quantification of the Akt protein obtained on the membrane was made with the ImageJ program. Results of Western blot analysis showed no significant change in expression level of Akt protein after treatment of CaCo-2 cells with SPD1 and SDP12 derivatives compared to untreated cells. Applied SPD1 on NCI-H358 caused moderately increased Akt protein expression in relation to the control cells. Obtained results suggest that Akt signaling pathway is not affected by tested compounds.

purine nucleoside derivatives, Akt expression, CaCo-2, NCI-H358

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