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Objective:To investigate the effect and mechanism of NACK knockdown on the proliferation and apoptosis of T-cell acute lymphoblastic leukemia (T-ALL) Jurkat cells. Methods:Lentivirus transfection technology was used to transfect Jurkat cells and knock down NACK gene.Real time fluorescent quantitative PCR and Western blot were used to detect the silencing efficiency of NACK gene.CCK-8 method and flow cytometry were used to detect the effects of NACK knockdown on the proliferation and apoptosis of Jurkat cells.The expressions of protein related with Notch1 pathway, such as Hes1 and c-Myc, were detected by Western blot. Results:After NACK-shRNA was successfully transfected into Jurkat cells by lentiviral vector, the expression of NACK mRNA and protein was reduced signi-ficantly ( P<0.05). Compared with the negative control group and the blank control group, the CCK-8 method showed that the cell proliferation in the experimental group was significantly inhibited [The inhibition rates of cell proliferation in the experimental group, negative control group and blank control group were (37.27±4.48)%, (4.25±2.10)% and (2.43±1.40)%, respectively]( F=132.640, P<0.05), and the flow cytometry test showed that the apoptosis in the experimental group increased significantly [The apoptosis rates of experimental group, negative control group and blank control group were (26.38±3.03)%, (6.07±2.61)% and (3.40±1.98)%, respectively]( F=90.534, P<0.05). Western blot results confirmed that the expression of Notch1 pathway-related proteins Hes1 and c-Myc was down-regulated compared with the negative control group and the blank control group, and the difference was statistically significant ( P<0.05). Conclusions:Targeting silent NACK can down-regulate the expression of Notch1 pathway-related proteins, which leads to the inhibition of Jurkat cell proliferation and increased apoptosis, thereby exerting its anti-T-ALL effect.
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Objective To study the effect of Tamibarotene on the SH-SY5Y cell proliferation inhibition ability and the mRNA and protein expressions of tyrosine kinase receptor a (TrkA) and N-myc (MYCN) in order to provide some experimental bases for the treatment of neuroblastoma.Methods The SH-SY5Y cells were treated with different concentrations of Am80 (0,10,20,40,80,160 μmol/L) for 48 h,then Cell Counting Kit-8 (CCK-8) was used to test the cell proliferation.Reverse transcription PCR(RT-PCR) and Western blot were used to test the mRNA and protein expressions of TrkA and MYCN at 48 hours.Results When the concentration was 10 μmol/L,Am80 had no significant inhibitory effect on SH-SY5Y cells [(3.51 ± 1.68)％,inhibition ratio ＜ 5 ％];but when the concentration was 20 μmol/L,it showed weak inhibition [(9.60 ± 1.97) ％,inhibition ratio ＜ 10％].The inhibition rate of SH-SY5Y cell proliferation[(57.43 ± 4.95)％] was significantly enhanced at Am80 with a concentration of 80 μmol/L.The concentrations of Am80 could effectively inhibit SH-SY5Y cell proliferation in a dose-dependent manner(P ＜0.05).The expression of TrkA increased with the increase of Am80 concentration.Am80 significantly decreased the expression of MYCN in SH-SY5Y cells(10 μmol/L:0.65 ±0.05 vs.20 μmol/L:0.36 ±0.06),and the difference was statistically significant(P ＜ 0.05).Conclusions It is suggested that Am80 can effectively inhibit SH-SY5Y cell proliferation in a concentration-dependent manner.The underlying mechanism involves increasing the expression of TrkA by down-regulation of MYCN.
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Objective@#To investigate the effects of Tiaozhi mixture on lipid metabolism disorder in patients with chronic hepatitis C virus infection and fatty liver.@*Methods@#The clinical data of 122 cases of patients with chronic hepatitis C virus infection and fatty liver admitted to our hospital from January 2016 to January 2017 were retrospectively analyzed. Control group was only given antiviral treatment, and observation group was given antiviral treatment and Tiaozhi mixture. The blood lipids parameters, decrease of HCV RNA load, negative conversion rate of serum anti-HCV.@*Results@#After treatment, the levels of blood lipids, i. e., triglycerides (TG), total cholesterol (TC) and low density lipoprotein-cholesterol (LDL-C) were decreased significantly in the two groups, and the decreases of blood lipid levels of TG, TC and LDL-C in observation group were greater than those in control group. The level of high density lipoprotein-colesterol (HDL-C) was increased significantly in observation group, and the level of HDL-C in control group was basically unchanged. The liver function tests for alanine aminotransferase (ALT) and total bile acid (TBA) levels were all decreased effectively, and the decrease in observation group was greater than that in control group, and the aspartate aminotransferase (AST) level in the two groups was reduced, but there was no significant difference between the two groups. The decrease of HCV RNA load and negative conversion rate of serum anti-HCV in observation group were higher than those in control group. There was no significant difference in the incidence rate of adverse reactions between the two groups after treatment(χ2=0.000, P=1.000).@*Conclusions@#In the clinical treatment of chronic hepatitis C virus infection with fatty liver, combined Tiaozhi mixture can effectively increase the antiviral efficacy, reduce the possibility of adverse reactions, and help patients with liver function recovery, and it has high safety. Thus it is worthy of clinical promotion.
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Objective To study the effect of tetramethylpyrazine injection on proliferation and apoptosis of human acute lymphoblastic leukemia (ALL) cell line Jurkat and the relevant molecular mechanisms.Methods Cells were treated with tetramethylpyrazine injection at various concentrations (0,0.25,0.50,0.75,1.00,1.25 and 1.50 mg/ml),CCK-8 method was used to detect the inhibition rates at 24,48 and 72 h.After cells were treated with different concentrations of tetramethylpyrazine injection (0,0.50,1.00 and 1.50 mg/ml) for 48 h,the cell cycle and apoptosis were detected by flow cytometry,and the Aurora-B and Survivin protein expression were analyzed by Western blot.Results Compared with the control group (cells without tetramethylpyrazine injection treatment),various concentrations of tetramethylpyrazine injection could effectively inhibit Jurkat cells proliferation in a time-and dose-dependent manner (P ＜ 0.05),and IC50 at 24 h,48 h and 72 h after treatment were (1.33±0.16),(0.91±0.10) and (0.67±0.11) mg/ml,respectively.After cells were treated with tetramethylpyrazine injection at different concentrations (0.5,1.0 and 1.5 mg/ml) for 48 h,the number of treated cells in G2/M phase was increased,but that in S phase was decreased,and the apoptosis rates were significantly higher than that of control group,with a dose dependence (P ＜ 0.05).The results of Western blot showed that the expression levels of Aurora-B and Survivin in treated cells were lower than that of control group,also with a dose dependence (P ＜ 0.05).Conclusions Tetramethylpyrazine injection can effectively inhibit proliferation and induce apoptosis of Jurkat cells in vitro,and its underlying mechanisms involve with down-regulation of the Aurora-B and Survivin protein expression.
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The SF3B1 gene encodes subunit 1 of the splicing factor 3b,which is a core component of the U2 small nuclear ribonucleoprotein and plays an important role in the process of RNA splicing. Abnormal splicing caused by SF3B1 mutations are associated with hematological malignancies,particularly with myelodys-plastic syndrome,refractory anemia with ring sideroblasts associated with marked thrombocytosis and chronic lymphocytic leukemia( CLL) . In myelodysplastic syndrome and refractory anemia with ring sideroblasts associat-ed with marked thrombocytosis,SF3B1 mutations are bond up with favorable prognosis and strongly with ring sideroblasts. But in CLL,SF3B1 mutations are factors of poor prognosis.