ABSTRACT
OBJECTIVE@#To investigate the pro-apoptotic effect and mechanism of miR-30a overexpression on chronic myeloid leukemia K562 cells.@*METHODS@#The k562 cells were transfected with the recombinant plasmid pEGFP-pre-miR-30a, the real-time quantitative PCR was used to detect the level of miR-30a and BCR/ABL, and then the cell apoptosis was assessed by flow cytometry with AnnexinV-FITC/PI double staining. Western blot was used to detect the expression of BCR/ABL protein,apoptosis-related protein BCL-2 and BAX, PTEN, AKT and p-AKT.@*RESULTS@#Sequencing and digestion map indicated that the recombinant plasmid was constructed successfully. Compared with 2 control groups, the miR-30a expression in k562 cells transfected with recombinant plasmid pEGFP-pre-miR-30a was obviously up-regulated. The expression of BCR/ABL mRNA and BCR/ABL protein was both significantly down-regulated. Apoptotic rate was significantly enhanced (both P<0.05),and the expression of anti-apoptotic protein BCL-2 was down-regulated while the expression of pro-apoptotic protein BAX was up-regulated. The level of PTEN was significantly up-regulated in omparison with control groups,no variation was found in total AKT, but the expression of p-AKT was down-regulated.@*CONCLUSION@#The overexpression of miR-30a is abled to down-regulate the level of BCR/ABL mRNA and BCR/ABL protein, and increase apoptotic rate, its mechanism may be related with inhibition of the activity of BCR/ABL-PTEN/AKT signaling pathway.
Subject(s)
Humans , Apoptosis , Cell Proliferation , Fusion Proteins, bcr-abl , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , MicroRNAs , GeneticsABSTRACT
To study the chemical constituents of Cymbopogon citratus, isolation and purification of constituents were carried out on silica gel, Sephadex LH-20 and prepatative HPLC. The structures of the compounds were identified by physicchemical properties and spectral data analysis. Eight compounds were isolated and identified as 3beta-methoxy lanosta-9(11)-en-27-ol (1), 3beta-hydroxylanosta-9 (11)-en (2), (24S) -3beta-methoxylanosta-9(11), 25-dien-24-ol (3), 8-hydroxyl-neo-menthol (4), (2E)-3,7-dimethyl-2,7-octadiene-1, 6-diol (5), (+)-citronellol (6), 7-hydroxymenthol (7) and ethyl nonadecanoate(8). Compounds 1 is a new one. Compounds 2-3 are obtained from C. citratus for the first time.