RUNX1 gene mutations are associated with adverse prognosis of patients with acute myeloidleukemia / 南方医科大学学报

OBJECTIVE@#To explore the rate and distribution of Runt- related transcription factor 1 (RUNX1) gene mutations in patients with acute myeloid leukemia (AML) and the correlation of these mutations with the clinical characteristics and survival outcomes of the patients.@*METHODS@#The genomic DNA extracted from the bone marrow of 158 patients with newly diagnosed AML for PCR amplification of RUNX1 gene and sequence analysis to identify the mutations. The mutations of ASXL1, DNMT3A, TET2, FLT3, CEBPA, NPM1, IDH2, NRAS and c-KIT genes were also examined to analyze their association with RUNX1 gene mutations.@*RESULTS@#Among the 158 AML patients, 19 (12.0%) were found to have RUNX1 mutations in A166G (9 cases), A142T (6 cases) and A162L (4 cases). RUNX1 mutations were more frequent in elderly patients (@*CONCLUSIONS@#RUNX1 gene mutations are associated with an adverse prognosis of patients with AML.

Effect of Artesunate on Transferrin Receptor in K562/ADM Cells / 现代生物医学进展

Objective:To investigate the effect of Artesunate (Art) on the expression of transferrin receptor (TtR)in K562/ADM cells.Methods:The drug-resistant K562/ADM cells were cultured with 1000 ng/mL doxorubicin for two weeks followed by Artesunate treatment with different concentrations (12.5 μg/mL,25μg/mL and 50 μg/mL) or different time (12 h,24 h,36 h,and 48 h).The content of transferrin receptor in K562/ADM cells was determined by flow cytometry.The effect of Artesunate on the expression of transferrin receptor protein in K562/ADM cells was measured by Westem blot.Cell counting kit-8 (CCK-8) assay was used to evaluate inhibitory effect of Art combined with doxorubicin (ADM) in K562/ADM cells.The reversal index was defined as the IC50 of the experimental group divided by the IC50 of the control group in K562/ADM cells.Results:Art effectively decreased the content of transferrin receptor and the expression of transferrin receptor protein in K562/ADM cells in a dose-dependent manner.Moreover,Art also inhibited transferrin receptor protein expression in K562/ADM cells in a time-dependent manner.The different concentrations of Art(12.5 μg/mL,25μg/mL and 50 μg/mL) could induce reversal of drug-resistance with the reversal index being 1.38,2.12 and 2.95 times (P＜0.05).Art inhibited cell proliferation of K562/ADM cells,and the IC50 werel9.7 μmol/L.Conclusions:Art effectively down-regulated the transferrin receptor content as well as transferrin receptor protein expression in K562/ADM cells,which resulted in reversal of drug resistance of K562/ADM cells.Art also inhibited K562/ADM cells proliferation,which has great value in clinical treatment of leukemia.

Effects of SLeX on invasion and migration of HepG2 cells / 中国病理生理杂志

AIM: To investigate the expression and effects of sialyl Lewis X (SLeX) on the invasion and migration of human hepatocellular carcinoma HepG2 cells.METHODS: The expression of α1,3-fucosyltransferase VII (FUT7) in HepG2 cells and L-02 cells was detected by RT-qPCR and Western blot.The SLeX expression in HepG2 cells and L-02 cells was determined by Western blot and immunocytochemical staining.The invasion and migration abilities of the treated cells were evaluated by Transwell assay.RESULTS: The expression of FUT7 and SLeX in the HepG2 cells, but not in the L-02 cells, was observed.The invasion rates of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L were significantly decreased as compared with control group (P<0.05).The migration ability of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L was also significantly reduced as compared with control group (P<0.05).The invasion rate and migratory cell number were significantly different between any 2 groups in the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L (P<0.05).CONCLUSION: HepG2 cells express SLeX.SLeX is closely related to the migration and invasion abilities of the HepG2 cells.

Potentiation of cisplatin induced apoptosis by low molecular weight heparin in human hepatocellular carcinoma cell / 中南大学学报(医学版)

OBJECTIVE@#To explore the effects of low molecular weight heparin (LMWH) on cisplatin (DDP)- induced apoptosis in human hepatocellular carcinoma cell and the underlying mechanisms.
@*METHODS@#Hepatocellular carcinoma SMMC-7721 cells were divided into a control group, a LMWH group, a DDP group and a LMWH plus DDP group. The effect of the drugs on proliferation of hepatocellular carcinoma SMMC-7721 cells were evaluated by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide (MTT) assay, and the apoptosis were detected by acridine orange/ethidium bromide (AO/EB) double fluorescence method and flow cytometry method. The expression levels of apoptosis-related protein Fas, Bcl-2 and Bax were detected by quantitative real-time PCR and Western blot.
@*RESULTS@#Compared with the control group, the proliferation of SMMC-7721 cells was inhibited in the DDP group and the LMWH plus DDP group (both P0.05). The Fas level was increased obviously (P0.05). Compared with the control group and the DDP group, the Bcl-2 level was reduced significantly in the LMWH plus DDP group (both P0.05).
@*CONCLUSION@#LMWH can enhance the cisplatin-induced apoptosis in SMMC-7721 cells, which might be related to activation of mitochondrial pathway.

Effect of protein kinase Cθ on the regulation of L-selectin expression of human γδT cells / 中华微生物学和免疫学杂志

Objective To investigate the role of PKCθ signal pathway on regulation of L-selectin (CD62L) expression in human activated γδT cells. Methods Human peripheral blood mononuclear cells (PBMC) were stimulated with Mycobacterium tuberculosis antigen (Mtb-Ag) and cultured for 6-8 d to generate Mtb-Ag activated T cells(MtbAT) as γδT cells enrichment T cell line. The MtbAT were stimulated with PMA or PMA + IMN for 3, 6, 12 and 24 h, respectively, or MtbAT cultured for 8 d were stimulated with Mtb-Ag, or PMA, with or without PKC0 inhibitor Rottlerin for 4 h. After the treated cells stained with fluo-rescent labeled monoclonal antibodies, the expression of CD62L on γδT cells were measured by flow cytome-try (FCM ). Results The expression of CD62L on γδT cells cultured for 6-8 d were 75.0%-87.0%. Decrease of CD62L from the surface of γδT cells by 3 h to 12 h after exposure to PMA (42.3% to 23.5% ), but CD62L expression increased to 53.2% when γδT cells were exposed to PMA for 24 h. The expression of CD62L of γδT cells decreased to 52.1% and 39.3% respectively when γδT cells were exposed to PMA + IMN for 3 h and 6 h. After treated with PMA + IMN for 12 h and 24 h, the expression of CD62L were 52.9% and 35. 3% respectively. The CD62L expression of γδT cells treated with PMA and Rottlerin (47.9%) were higher than that treated with PMA alone (31.8%). After Mtb-Ag restimulated MtbAT for 4 h, the CD62L level of γδT cells decreased from 70.0% to 54.8%, Rottlerin could inhibite Mtb-Ag down regulation CD62L level of γδT cells (63.1%). Conclusion The CD62L expression of γδT cells could be ingibited partly by the inhibitor of PKCθ signal pathway may regulate L-selectin (CD62L) expression of activated human γδT cells.