香菇提取物对白血病细胞HL-60增殖的抑制作用

香菇是全球第二大人工种植最为普遍的食用菌。研究表明,香菇具有抗癌、抗氧化、抗菌及降血脂等活性,目前大多数研究集中于香菇子实体的抗癌活性。在对香菇在固体培养基上进行培养的过程中,培养基中发现菌丝分泌一种棕色物质,猜测这些棕色物质可能具有某些生物活性。本研究的目的在于探讨这些棕色物质是否对急性早幼粒细胞性白血病细胞HL-60的增殖具有抑制作用。香菇在固体培养基上培养3￣4周后,通过离心和冷冻干燥得到棕色物质,然后进行一系列的分离纯化。通过台盼蓝染色排除法测定了得到的各种组分对HL-60增殖的抑制作用,通过荧光显微镜双染色法测定了各组分对HL-60细胞凋亡的诱导作用。结果表明,作用72h后,粗提物、透析所得低分子量组分和大孔树脂分离所得低极性组分对HL-60的细胞增殖具有明显的抑制作用,IC50分别为(135±0.59),(102±0.66)和(23±0.45)μg/mL。同时,粗提物中的活性成分为低极性物质,其分子量小于10000D,并非香菇多糖。荧光显微镜双染色结果表明,作用12h后,粗提物诱导HL-60细胞的凋亡指数由32%提高至64%。结果表明,香菇提取物对于HL-60细胞的增殖具有显著的抑制作用。     

Fibrinolytic activity and its mechanisms in various leukemic cell lines

AIM:To study the relation between expression of uPAR and annexinⅡ and fibrinolytic activity in various leukemic cell lines. METHODS:The plasma activity was measured under the reaction between cells of NB4, SHI-1, K562, Jurkat, Raji and plaminogen by chromogenic assay. The protein expressions of uPAR and annexinⅡin cells of NB4, SHI-1, K562, Jurkat, Raji were detected by flow cytometry method. The mRNA expressions of uPAR and annexinⅡin cells of NB4, SHI-1, K562, Jurkat, Raji were detected by RT-PCR. RESULTS:The plasma activity in SHI-1 cells and NB4 cells were higher obviously than that in Raji, K562 and Jurkat cells. The protein expression ratio of uPAR and annexinⅡ in NB4 cells were (13.15±1.61)% and (95.97±1.19)%， respectively, they were (99.00±0.26)%, (90.35±2.15)% respectively in SHI-1 cells, and they were lower in K562, Jurkat, Raji cells. The expression of annexinⅡ mRNA in NB4 cells was higher than that in SHI-1 cells, and they were undectectable in K562 and Jurkat cells. The expression of uPAR mRNA in NB4 and SHI-1 cells were higher than that in Jurkat and K562 cells. The expression of uPAR mRNA in Raji cells was undectectable. CONCLUSION:The primary hyperfibrinolysis in leucocythemia cells was observed, and relation was closely with the expression of annexinⅡ. It might be the main reason for bleeding and disseminated intravascular coagulation in patients with acute promyelocytic leukemia and acute monocytic leukemia.     

Alteration of circulating endothelial cells from acute promyelocytic leukemia patients before and after treatment and its influential factors

AIM: To determine the biological feature of circulating endothelial cells (CECs) in acute promyelocytic leukemia (APL) patients before and after treatment, and to analyze the relationship between CECs and the clinical characteristics. METHODS: The CECs were sorted from peripheral blood by magnetic-activated cell sorting and then counted by 3-color flow cytometry. The cells were identified by immunofluorescence staining for the expression of CD146, CD31, CD144, VEGFR-2, CD45 and CD133. The CECs were cultured in vitro, and the tube formation and colony-forming rate were determined. RESULTS: Increased quantity of CECs was observed in CD34 positive group and group with WBC>10×10⁹/L (P<0.05). The quantity of CECs had a significant difference among low risk, medium risk and high risk groups (P<0.05). The positive rate of CD133 and quantity of CECs significantly reduced in 32 APL patients when they gain complete remission after treatment (P<0.05). The amount of tube formation and colony-forming rate were significantly reduced after treatment (P<0.05). The ratio of CECs quantity from APL patients after treatment to that before treatment had a negative correlation with arsenic concentration in urine on day 7 during As₂O₃ treatment (P<0.05). CONCLUSION: Accurately counting CECs may be helpful for evaluating prognosis and designing treatment strategy.     

Effect of glutamine deprivation on growth and differentiation of primary APL cells

AIM:To investigate the effect of Gln deprivation on growth and differentiation of primary APL cells. METHODS: The primary APL cells were collected from 18 cases of APL patients’ periphery blood, the patients had not been treated. The cells were incubated in RPMI 1640 without or with different contents of glutamine and with 10% fetal calf sera, at 37℃,5% CO 2 and saturation humidity for 4 days.The initial living cell density was 5×10⁸/L. After 4 days incubation,the cells were counted,collected, smeard and stained with Wright-Giemsa, DNA, POX, NAE and NaF inhibition,gulping ink and NBT etc. cytochemical technology. The stained cells were investigated under oil immersion lens. RESULTS: The living cell density became (54.28±4.28)% of the initial in glutamine deprivation group, but the living cell density in control with 4 mmol/L Gln was (108.56±12.27 )% of the initial ( P <0.01) after 4 days incubation. The cells differentiated into mature granulocytes in glutamine deprivation group.CONCLUSION: Deprivation of glutamine inhibited primary APL cell growth and induced cell differentiation toward mature granulocyte.