Effect of interferon on HEL cell apoptosis and JAK2 V617F mutation gene expression / 实用医学杂志

Objective To observe the effect of Interferon-α2b on HEL cells (human erythroleukemia cell line) growth, apoptosis and JAK2 V617F mutation gene expression. Methods HEL cells were placed in RPMI1640 containing 10% FBS and incubated in a cell incubator. Cells in the logarithmic growth phasem were collected, adjusting the cell density to 1 × 105/mL for experimental research. The interferon concentration in five groups were 0, 5 × 105, 10 × 105, 50 × 105, 100 × 105 U/L, with different incubation time (0, 24, 72, 120 h), respectively. The cell growth status in different groups was observed in the inverted optical microscope; MTT was used to detect the inhibition of interferon on HEL cell proliferation. Cell apoptosis was detected by flow cytometry. Fluorescence quantitative PCR was used to detect the mutation gene of JAK2 V617F expression. Results Inhibition rates of Interferon on the HEL cell proliferation in 5 × 105 U/L, 10 × 105 U/L, 50 × 105 U/L, 100 × 105 U/L groups were 18.57%, 25.10%, 42.10%, 57.00%, respectively. JAK2 V617F/GAPDH by fluorescence quantitative was 1.556, 1.213, 0.870 respectively under the concentration of interferon 100 × 105 U/L for 24, 72, 120 h. Conclusions Interferon-α2b can inhibit HEL cells proliferation and induce HEL cells apoptosis. Increasing concentration of interferon increases HEL cell apoptosis rate. Interferon can inhibit JAK2 V617F expression of HEL cells in a dose-dependent manner.

The influence of heparin on the apoptosis and proliferation for K562 cells induced by vincristine / 白血病·淋巴瘤

Objective To investigate the effect of heparin on the apoptosis and proliferation of human myeloid leukemia cell line K562 induced by vincristine.Methods K562 cells were pretreated by heparin for 1h,then cultured with 0.05 mg/L vincristine in 37℃ 5% CO2 for 24 h.Apoptosis of KS62 cells was evaluated by Hoechst 33342 staining,flow cytometer and DNA agarose gel electrophoresis after culture for 24 hours.The effect of heparin on KS62 cell proliferation and toxicitv was determined by Trypan blue staining and MTT assay.Results In the apeptosis induced group,the apeptosis rate was 40.10% dected by Hoechst 33342 fluorescence staining.The hepafin in different concentrations was found to be able to inhibit the apoptosis of K562 cells triggered by vincristine and the apoptosis rate was 32.47%,29.7%,25.5%,19.53% in the heparin groups of 25,50,100,200 U/ml,respectively.The apeptosis rate was significantly lower in the apeptosis induced group than in the heparin groups of 25,50,100,200 U/ml(P＜0.05).The typical DNA ladder could be found in the apoptosis-induced group,and the DNA ladder gradually disappeared along with the increase of heparin(5～200 U/ml).The sub-G1 peak of K562 cells could be found in the induced group by FACS and the apoptosis rate was 21.61%.In the heparin groups of 25,50,100,200 U/ml,the sub-G1 Peak of K562 cells gradually dropped and the apoptosis rate was 13.64%,11.75%,8.59%,6.03%(P＜0.05),respectively.After K562 cells were incubated with different hepafin concentrations(5～200 U/ml)for 24 hours,there was no difference compared with the normal control group in both the total live cell numbers and the cell proliferation rate measured by trypan blue staining and MTT assay(P＞0.05).Conclusion The results suggested that heparin had no influence on KS62 cell toxicity and proliferation,but may inhibit the apoptosis of KS62 cells induced by vincristine.

Promotion of Angelica polysaccharide on inducing chronic myelocytic leukemia cells into dendritic cells / 中草药

Object To investigate the effect of Angelica polysaccharide (APS) on the induction of chronic myelocytic leukemia cells into chronic myelocytic leukaemia dendritic cells (CML-DCs). Methods Bone marrow monocytes from CML patients were cultured in GM-CSF/IL-4 or in GM-CSF/IL-4 combined with APS in each concentration (50, 100, 200 mg/L), respectively. The morphotype of CML-DCs was identified by optical microscope or electron microscope, CML-DCs viability was calculated by Trypan Blue exclusion. The phenotype of CML-DCs (CD 80, CD 86, and CD 83) was identified by flow cytometry. The capability of stimulating auto-lymphocyte or allo-lymphocyte proliferation was tested with mixed leukocyte reaction (MLR). Results Bone marrow monocytes from CML patients, which were cultured in GM-CSF/IL-4 or in GM-CSF/IL-4/APS showed typical morphotype and expressed the high level phenotype of CML-DCs. The capability of proliferation and the survival rate of CML-DCs were enhanced markedly and the expression of CD 83, CD 80, and CD 86 on CML-DCs were significantly increased when CML-DCs were cultured in GM-CSF/IL-4/APS. The capability of stimulating lymphocyte proliferation was more competent in 100 mg/L APS group. Conclusion The expression of CD 83, CD 80, and CD 86 on CML-DCs cultured in GM-CSF/IL-4/APS is significantly higher than those in GM-CSF/IL-4. The capability of CML-DCs of stimulating lymphocyte proliferation is more potential in GM-CSF/IL-4/APS than in GM-CSF/IL-4. APS can promote the induction and mature of CML-DCs cultured in IL-4 and GM-CSF.