Effect of Triptonide on Proliferation and Apoptosis of SHI-1 Cells / 中国实验方剂学杂志

Objective:This study aims to investigate the effect of triptonide （TN） on proliferation， cell cycle， apoptosis and expressions of apoptosis-related proteins of human acute monocytic leukemia（AML） cell line SHI-1， and to explore its possible mechanism of action. Method:The thiazolyl blue （MTT） colorimetric assay was applied to detect the inhibitory effect of 20，40，80，160，320 nmol·L^-1 triptonide on the proliferation of SHI-1 cells for 48， 72 h. Changes in SHI-1 cell cycle before and after triptonide treatment were detected by flow cytometry propidium iodide （PI） simple staining， and changes in SHI-1 cell apoptosis before and after triptonide treatment were detected by flow cytometry with AnnexinV/PI double staining. Western blot was applied to detect the protein expression of cysteine protease （Caspase）-3， Caspase-8 and nuclear transcription factor kappaB（NF-<italic>κ</italic>B） in SHI-1 cells before and after treatment with 80， 160 nmol·L^-1triptonide. Result:Compared with the blank group， 40，80，160，320 nmol·L^-1 triptonide significantly inhibited the proliferation of SHI-1 cells（<italic>P</italic><0.01） in a dose-dependent manner for 48， 72 h， while 160， 320 nmol·L^-1 triptonide induced apoptosis of SHI-1 cells（<italic>P</italic><0.01） for 48， 72 h， and 160 nmol·L^-1 triptonide could decrease the S phase ratio of SHI-1 cells（<italic>P</italic><0.01）. In addition， compared with the blank group， 80，160 nmol·L^-1 triptonide induced the downregulation of NF-<italic>κ</italic>B significantly（<italic>P</italic><0.01）， 160 nmol·L^-1 triptonide induced the downregulation of Caspase-3， Caspase-8 significantly（<italic>P</italic><0.01）. Conclusion:Triptonide can inhibit the proliferation and induce apoptosis <italic>in vitro</italic> of SHI-1 cells， which may be related to the reduction of the cells in S phase proportion by triptonide， and the downregulation of the expression levels of Caspase-3， Caspase-8 and NF-<italic>κ</italic>B proteins.

Effect of LSD1 knock-down by small hairpin RNA on the apoptosis and cell cycle of human acute myelogenous leukemia cells / 实用医学杂志

Objective To investigate the effect of down-regulation of lysine specific demethylase 1 (LSD1) by shRNA on the apoptosis and cell cycle of human acute myelogenous leukemia cells.Methods The lentiviral vector-mediated LSD1-shRNA was transfected into human acute promyelocytic leukemia HL-60 cells and acute monocytic leukemia SHI-1 cells.The expressions of LSD1 mRNA and protein were examined by real time quantitative PCR and Western blot,respectively.The flow cytometry was applied to detect the apoptosis and cell cycle distribution after AnnexinV-PE/7-AAD and PI dying,respectively.Results The expressions of LSD1 mRNA and protein in HL-60 and SHI-1 LSD1-shRNA group were significantly decreased compared with the blank control group and the negative shRNA group (P ＜ 0.01,respectively).The apoptosis levels of HL-60 and SHI-1 cells were significantly increased after knockdown of LSD1 (P ＜ 0.01).Moreover,the cell cycle distribution in the G0/G1 phases was also significantly increased(P ＜ 0.01).Conclusion LSDI-shRNA promotes cell apoptosis and increases the percentage of cells in the G0/G1 phases of human acute myelogenous leukemia cells.

Analysis of VOCs in the headspace of SHI-1 acute mononuclear leukemia cell / 安徽医科大学学报

Objective To find out the impact of SHI-1 cells on VOCs in the headspace, and to assess the feasibili-ty of VOCs analysis in leukemia patients or with other tumors in the differential diagnosis by determing VOCs in the headspace of acute mononuclear leukemia cells(SHI-1). Methods The air samples from the headspace of SHI-1, Human macrophages cells and medium control were collected by syringes, and then determined by means of solid phase microextraction-gas chromatography/mass spectrometry ( SPME-GC/MS ) in order to have a better under-standing of the concentration distributions and changes of VOCs in SHI-1 cells' headspace. Results Using Mann-Whitney U test, we found that eight kinds of VOCs were different among them. They were 2,4-dimethyl heptanes, 4-methyl octane, chloroform, benzene, 3,7-dimethyl dodecanese, hexanol, cyclohexanol, cetane. And alkane, benzene were significantly higher than the control group ( P<0.05 ) , alcohol decreased significantly ( P<0.05 ) . Conclusion SHI-1 cells have an impact on the concentration distributions of VOCs in their headspace. The most outstanding VOCs are alkanes, alcohol and benzene compared with the blank control group. They may be potential markers characteristic of leukemia cells.

Experimental study on the differentiation of SHI-1 cells induced by puerariae radix flavones in vitro / 白血病·淋巴瘤

Objective To explore the possible effects on differentiation of SHI-1 cells induced by puerariae radix flavones(PRF)in vitro.Methods SHI-1 cells were treated with PRF in various concertration,then the inhibitory effects of cell proliferation were detected by MTT assay,the cell cycles were analyzed by flow cytometry(FCM),the cells reduction rates were detected by NBT reduction test,and the expression of CD11b and CD14 were tested by FCM.Results 10-50 μg/ml PRF could inhibit the proliferation of SHI-1 cells in a time-and dose-dependent manner,and the cell cycles were arrested in S phase.When SHI-1 cells were treated with 10,30 and 50 μg/ml PRF in 48 houres respectively,the NBT reduction rates of cells were increased in a dose-dependent with PRF(P＜0.05),and the expression of cells surface differentiation antigen CD14 was also increased along with the concentration of PRF.Conclusion The SHI-1 cells could be induced to differentiation partially after treated with 10,30 and 50 μg/ml PRF in vitro.

Effect of artesunat on the expression of vascular endothelial growth factor and vascular endothelial growth factor receptors in SHI-1 cell line / 白血病·淋巴瘤

ObjectiveTo investigate the effect of artesunate on the expression of vascular endothlial growth factors(VEGF)and VEGFR in SHI-1 cell line.MethodsEnzyme-linked immunosorbent assay analysis was performed to detect the amount of VEGF in culture supernatants of SHI-1 cell in the condition of artesunate or not. The expression of VEGFR1 and VEGFR2 in SHI-1 cell in the condition of artesunate or not were detected by flow cytometry.ResultsWithout artesunate,the concentration of VEGF in the culture supernatant of SHI-1 cell were (980.3±2.2) pg/ml in 24 h and (982.4±2.3) pg/ml in 48 h. The expression of VEGFRI in SHI-1 cell were (6.40±3.11) ％ in 24 h and (6.45±2.85) ％ in 48 h. The expression of VEGFR2 in SHI-1 cell were (13.90±2.26) ％ in 24 h and (13.95±1.96) ％ in 48 h. With artesunate at 5, 10, 20 ng/ml, the concentration of VEGF in culture supematant of SHI-1 cell were (234.6±1.8)pg/ml, (114.9±1.6)pg/ml, (108.8±1.5) pg/ml in 24 h and (62.3±1.7) pg/ml, (60.9±1.6) pg/ml, (32.7±1.7) pg/ml in 48 h, respectively. The levels of VEGF in SHI-1 cells treated with artesunate at different concentrations decreased significantly (P ＜0.05).There was significant difference between 24 hours group and 48 hours group(P ＜0.05).The expression of VEGFR1 in SHI-1 cell were (4.30±2.21) ％, (4.20±1.37) ％, (3.90±1.86) ％ in 24 h and (3.80±2.87) ％, (3.60±1.73) ％, (3.00±1.82) ％ in 48 h, respectively. The expression of VEGFR1 in SHI-1 cell treated with artesunate at different concentrations were not significantly different (P ＞0.05). No significant difference between 24 hours group and 48 hours group was observed (P ＞0.05). VEGFR2 expression of SHI-1 cell were(4.40±1.15) ％, (3.10±0.68) ％, (1.10±0.72) ％ in 24 h and (3.00±1.68) ％, (2.20±0.93) ％, (0.60±0.92) ％ in 48 h, respectively. The results indicated that the expression of VEGFR2 in SHI-1 cells treated with artesunate at different concentrations reduced significantly (P ＜0.05),but there was no significant difference between 24 h group and 48 h group (P ＞0.05). ConclusionThe concentration of VEGF in SHI-1 cell was high, and artesunate can down-regulate the expression of VEGF and VEGFR2,but the effect of artesunate on the VEGFR1 was not significant.