ABSTRACT
OBJECTIVE@#To construct a K562 and adriamycin-resistant K562 (KAR) cell line with stably down-regulation of NCL (nucleolin) expression, and to investigate the effect of NCL down-regulation on the drug resistance in K562 and KAR cells.@*METHODS@#K562 and KAR cells were infected with lentivirus, and stably transfected cell clones were obtained by puromycin screening. The cell proliferation was detected by MTS assay, the cell apoptosis was detected by flow cytometry, and the expression level of drug resistance related genes was detected by real-time PCR.@*RESULTS@#The K562 and KAR cells with stable down-regulation of NCL were successfully constructed. Compared with the control group, the proliferation of K562 and KAR cells with down-regulating NCL expression decreased significantly (P ＜0.05), the apoptosis of cells increased significantly (P ＜0.05), and cell resistance to adriamycin was down-regulated.@*CONCLUSION@#Inhibition of NCL expression may increase the sensitivity of cells to adriamycin, which may be related with the promotion of apoptosis of K562 and KAR cells.
Subject(s)
Humans , Apoptosis , Doxorubicin , Drug Resistance, Neoplasm , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Phosphoproteins , RNA-Binding ProteinsABSTRACT
OBJECTIVE@#To investigate the role of nucleophosmin (NPM) in the proliferation of chronic myeloid leukemia cells (K562 cells) and its mechanism by RNAi technology.@*METHODS@#shRNA was used to inhibit the expression of NPM. The expression of NPM gene was detected by real-time quantitative PCR. The effect of inhibiting NPM gene on cell proliferation was detected by MTS assay. Change of cell cycle was detected by flow cytometry. Western blot was used to detect the expression of cell cycle-related proteins.@*RESULTS@#The shRNA lentiviral vector targeting at NPM gene was successfully constructed and used to transfect the K562 cells. The results showed that compared with the control groups, suppression of NPM gene expression in K562 cells could inhibit the cell proliferation and decrease the cell colony formation. Moreover, interference of NPM gene could prolong G/G phase and arrest cell cycle, which may be related to the down-regulation of NPM gene expression and activation of p21 protein expression, thereby inhibited the formation of CDK2/ Cyclin E complex.@*CONCLUSION@#Down-regulation of NPM gene expression in K562 cells can induce cell cycle arrest and inhibit cell proliferation.
Subject(s)
Humans , Apoptosis , Cell Proliferation , Gene Knockdown Techniques , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Nuclear ProteinsABSTRACT
<p><b>OBJECTIVE</b>To explore the changes of metabolites in brain after treatment by analysis on 1H-MRS examination of the hippocampus and prefrontal lobe in the healthy volunteers and the depression patients.</p><p><b>METHODS</b>Seventy-five cases of mild and moderate depression were randomly divided into groups A, B and C, 25 cases in each group. The group A was treated with oral administration of Prozac capsule, 20 mg/d; the group B by electroacupuncture for 30-40 min and the needle was retained for 1 h, once each day, with main points Baihui (GV 20), Yin-tang (GV 29) and adjuvant acupoints selected; the group C by combination of the treatment methods in the groups A and B. They were treated for 6 weeks. Use PROBE-J sequence at the MRI system on Single Voxel of ROI of each lateral of hippocampus and frontal lobe in the depression patients of the 3 groups. Compare the differences of N-acetylasp artate/creatine (NAA/Cr) and choline/creatine (Cho/Cr) between the healthy volunteers and the patients before and after treatment.</p><p><b>RESULTS</b>Before treatment, NAA/Cr in the bilateral hippocampus decreased in the 3 groups as compared with the control group, and after treatment, NAA/Cr in the bilateral hippocampus of the group B and in the right hippocampus of the group C increased compared with that before treatment (P < 0.05), and NAA/Cr in the left hippocampus of the group C significantly increased as compared with that before treatment (P < 0.01). Before treatment, Cho/Cr in the bilateral prefrontal lobe in the 3 groups increased as compared with that in the control group (P < 0.05), after treatment, Cho/Cr in the bilateral prefrontal lobes of the groups A, B and C significantly decreased as compared with that before treatment (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>There are differences in contents of metabolites in corresponding parts in bilateral frontal lobes and hippocampus between the depression patient and healthy person.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Choline , Metabolism , Creatine , Metabolism , Depression , Diagnostic Imaging , Metabolism , Therapeutics , Electroacupuncture , Frontal Lobe , Diagnostic Imaging , Metabolism , Hippocampus , Diagnostic Imaging , Metabolism , Magnetic Resonance Imaging , RadiographyABSTRACT
We isolated 61 endophyte isolates from the bark of 4 plants, Ginkgo biloba L, Albizzia julibrissin Durazz, Ailanthus sltissima (Mill) Swingle and Melia azedarach L. At the test concentration of 200 ?g/mL, higher than 50% of antitumor activities were demonstrated with crude extracts from 45.9% of fungal culture in MTT assay. Six isolates, YX5, YX17, YX36, KL1, CC1 and CC5, still showed higher cytotoxicity at 50 ?g/mL. No isolates from A. julibrissin had inhibitory effect towards EC109 at the test concentration of 50 ?g/mL; while about 15.8% of isolates from G. biloba were active. IC50 of the extract from the most active isolate YX5 against EC109, HONE1 and HeLa were 18.3 ?g/mL, 3.6 ?g/mL and 6.5 ?g/mL, respectively. Our results indicate that endophytes from G. biloba could be regarded as a potent source of antitumor drugs.