ABSTRACT
Objective To analyze the role of hDOT1L signaling pathway on the proliferation of human acute monocytic leukemia cell line THP-1 cell inhibited by demcitabine, and to explore the other effects except DNA demethylation and the mechanism of hDOT1L signaling pathways involved in leukemia. Methods Methyl thiazolyl tetrazolium (MTT) method was used to detect the influence of decitabine on THP-1 cell proliferation, and trypan blue anti-staining was applied to detect the effect of decitabine on THP-1 cell survival rate, reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression levels of hDOT1L, HOXA9, HOXA10, MLL-AF10 (MLLT10) mRNA, then the methylation level of H3K79 was detected by Western blotting assay. Results Compared with the control group, decitabine inhibited the proliferation of THP-1 cell in a time and dose-dependent manner. The inhibitory effect of 72 h was the highest, and the highest inhibition rate was 56.18%. In addition to 0.5 μmol/L in decitabine treatment group, the other groups were statistically significant (all P<0.05). There were high expressions of hDOT1L, HOXA9, HOXA10, MLLT10 mRNA and H3K79 hypermethylation in the THP-1 cell, meanwhile decitabine reduced the levels of hDOT1L mRNA after 48 h as well as HOXA9, HOXA10, MLLT10 mRNA, and there was a significant difference compared with the control group (all P < 0.01). Decitabine reduced obviously the methylation of H3K79 in cell after 48 h, especially the expression levels of bis and three methyl protein, and these differences were significant compared with the control group (all P< 0.01). Conclusion Decitabine plays its inhibitory effect on the proliferation of THP-1 cells probably through reducing the expression level of hDOT1L and H3K79 methylation level and decreasing the expression levels of key molecules, HOXA9, MLL-AF10, HOXA10, MLLT10 in the hDOT1L signal pathway.
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BACKGROUND:Cytotoxicity of graphene oxide to normal cel s is relatively low, but whether graphene oxide loaded with doxorubicin has some effects on malignant cel s is rarely reported. OBJECTIVE:To explore the cytotoxicity of graphene oxide loaded with doxorubicin on multiple myeloma cel s. METHODS:Multiple myeloma cel line RPMI8226 in logarithmic phase was selected, cultured and divided into four groups, including graphene oxide loaded with doxorubicin, doxorubicin and graphene oxide groups as wel as control group with no intervention. After 24 hours of culture, the cel activity was detected by cel counting kit-8 method, and the cel cycle and apoptosis were detected using flow cytometry. RESULTS AND CONCLUSION:Plump-shaped cel s with translucent and clear cytoplasm were found in the control group;cel s with relatively translucent cytoplasm, and even a few shrunken cel s appeared in the graphene oxide group;cel ular morphology was in a heterogeneity, apoptotic bodies appeared in the doxorubicin group;the cel s was significantly reduced in size, presenting more obvious shrinkage and apoptotic bodies in the group of graphene oxide loaded with doxorubicin. The cel survival rate in the graphene oxide loaded with doxorubicin, doxorubicin and graphene oxide groups was significantly lower than that in the control group (P<0.05), and this indicator was significantly lower in the group of graphene oxide loaded with doxorubicin than the graphene oxide group (P<0.05). The apoptotic rate in the group of graphene oxide loaded with doxorubicin and doxorubicin group was significantly higher than those in the graphene oxide and control groups (P<0.05), respectively. Additional y, there were no significant differences in the cel cycle among groups. These results show that graphene oxide loaded with doxorubicin has a stronger cytotoxicity, and can induce apoptosis in human multiple myeloma cel s.
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Objective To study the expression of PI3K and Akt mRNA in acute leukemia(AL) and chronic myelocytic leukemia (CML) and try to find out a relationship between the prognostic significance of acute leukemia and the expression level of PI3K and Akt. Methods The samples were collected from 63 ALpatients, 7 relapsed AL patients, 14 CML patients and 10 AL patients in complete remission(CR) patients, 11samples of normal controls(NC). The expression of PI3K, Akt mRNA were measured by semi-quantity reverse transcription polymerase chain reaction(RT-PCR). Results The expression of PI3K mRNA in AL and relapse group were significantly higher than that in normal control. In CML, the P13K mRNA expression level was higher than that in NC and CR. The similar results of the expression of Akt mRNA were observed in CML, NC and CR. At the same time, the expression level of Akt mRNA in relapse group was significantly higher than that in NC. The complete remission rate in PI3K (+) was lower than that in PI3K (-). Similar result was obtained in Akt (+) and Akt (-). Conclusion The PI3K/Akt pathway may contribute to the occurrence of leukemia. The patients with positive expression of the members of this pathway had a lower remission rate.
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Objective To investigate the expression of suppressor of cytokine signaling genes (SOCSs) and JAKs mRNA in the acute myloid leukemia(AML) patients. Methods The expression of SOCSs and JAKs mRNA as well as TYK2 in AML patients and healthy adults as normal contrals (NC) was measured with RT-PCR. Results The expression of SOCS 1,4,5 and 7 in AML patients was lower than those in normal control and AML with remis-sion (P<0.01),but the expression of SOCS 3 and 6 was higher than those in normal control and remission AML(P<0.01),however there was no significant difference in SOC2 between groups. The expressions of JAK2 ,JAK3 and TYK2 in AML were significantly higher than those in patients with remission and normal control(P<0.05). The ex-pression of JAK1 mRNA in relapsed AML was higher than that in normal control group(P<0.05),but the latter has no statistical significance between beginning treatment and normal group(P>0.05). Conclusion The deletion and degradion of SOCS 1,4,5 and 7 present in AML patients and JAKs expression is significantly increased, suggesting that both of them may co-participate in the pathogenesis of AML.