Expression Level and Correlation of miR-211, miR-155, C-myc in Acute T Lymphocytic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression and correlation of miR-211, miR-155, and C-myc in acute T lymphocytic leukemia (T-ALL), aiming to provide evidence for the diagnosis and treatment.@*METHODS@#A total of 96 T-ALL patients who were diagnosed and treated in People's Hospital of Zhengzhou from June 2014 to June 2017 were selected, and 69 healthy volunteers who had a physical examination were selected as control group in the same period. Real-time fluorescent quantitative PCR (RT-PCR) was used to determine the expression levels of miR-211, miR-155, and C-myc in peripheral blood mononuclear cells in each group. Kaplan-Meier was used to analyze the survival of T-ALL patients and correlation of miR-211, miR-155, and C-myc with prognosis. Pearson correlation analysis was used to evaluate the correlation of miR-211, miR-155, and C-myc with disease risk.@*RESULTS@#The expression levels of miR-211 mRNA, miR-155 mRNA, and C-myc mRNA in T-ALL group were higher than those in the control group (P<0.01), those in non-remission group were higher than those in remission group (P<0.01), and those in high-risk group were also higher than those in low-risk group and intermediate-risk group (P<0.01). The survival time of T-ALL patients with low miR-211 expression was longer than that with high miR-211 expression (P<0.01), that with low miR-155 expression was longer than that with high miR-155 expression (P<0.01), and that with low C-myc expression was also longer than that with high C-myc expression (P<0.01). The high expression of miR-211, miR-155, and C-myc was linearly positively correlated with high risk of disease (r=0.749, 0.781, 0.804).@*CONCLUSION@#The expressions of miR-211, miR-155, and C-myc are up-regulated in T-ALL patients, closely related to prognosis, and linearly positively correlated with disease risk.

The Relationship between PPP2R5C and Molt-4 Cell Viability, HSP90-GR Signal in Childhood Acute T Lymphocytic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of PPP2R5C to the activity of Molt-4 cells in childhood acute T lymphocytic leukemia and its mechanism.@*METHODS@#The small interfering RNA (siRNA) technology targeting PPP2R5C gene was used to down-regulate the expression of PPP2R5C in Molt-4 cells. At the same time, a blank control group, a negative control group and a 17-DMAG group were set up. The cells in the negative control group were transfected with siRNA-NC, the cells in 17-DMAG group were treated with the HSP90 inhibitor 17-DMAG at a final concentration of 6.4 μmol/L for 48 h. Real-time fluorescent quantitative PCR (RT-qPCR) and Western blot were used to detect transfection efficiency; CCK-8 method was used to detect the proliferation activity of the cells in each group, EdU was used to detect the proliferation level of the cells in each group, flow cytometry was used to detect the cell cycle distribution ratio of the cells in each group, Annexin V-FITC/PI staining was used to detect the apoptosis of the cell, RT-qPCR and Western blot were used to detect the expression changes of heat shock protein 90 (HSP90) and glucocorticoid receptor (GR) of the cells in each group.@*RESULTS@#After Molt-4 cells were transfected with siRNA-PPP2R5C, the expression of PPP2R5C mRNA and protein in the cells were down-regulated significantly compared with those in the blank control group and the si-NC group (P<0.05); compared with cells in the blank control group and the si-NC group, the proliferation activity of the cells in the siRNA-PPP2R5C group and the 17-DMAG group significantly decreased (P<0.05), and the rate of EdU positive cells was significantly reduced (P<0.05); the proportion of the cells in G1 phase decreased while the proportion of the cells in G2 phase increased (P<0.05), the apoptosis rate of the cells also increased significantly (P<0.05); in addition, the expression of PPP2R5C mRNA and protein of the cells in siRNA-PPP2R5C group was significantly down-regulated compared with those in the blank control group and si-NC group (P<0.05). The expressions of PPP2R5C mRNA and protein in the 17-DMAG group were also significantly down-regulated compared with those in the blank control group and si-NC group (P<0.05).@*CONCLUSION@#Down-regulation of PPP2R5C gene expression can inhibit Molt-4 cell activity in childhood acute T lymphocytic leukemia, block the cells in G2 phase, and promote cell apoptosis, the mechanism may be related to the inhibition of HSP90-GR signaling pathway.

Anti-tumor Efficacy and Safety of Zeylenone on Acute T Lymphoblastic Leukemia / 中国实验方剂学杂志

Objective:To investigate the anti-tumor efficacy, mechanism and safety of zeylenone on acute T lymphocytic leukemia. Method:In vitro, Molt-4 cells were treated with various concentrations of zeylenone (0.2, 0.4, 0.8, 1.6, 3.2 μmol·L-1) for 48 h, and the cell viability was measured with cell counting kit-8 (CCK-8) assay. nonobese diabetic-severce combined immunodeficient mice（NOD/SCID） mice were randomly divided into six groups: normal group, model group, vincristine group (1 mg·kg-1), low-dose zeylenone group (12.5 mg·kg-1), medium-dose zeylenone group (25 mg·kg-1), high-dose zeylenone group (50 mg·kg-1). With the exception of normal group, mice were pre-irradiated with 60Co and inoculated subcutaneously with Molt-4 cells to establish the Molt-4 xenograft model. Then NOD/SCID mice were sacrificed after 13 days of administration. The tumor inhibition rates, relative tumor growth rates and organ indexes were calculated. Hematoxylin and eosin (HE) staining was used to observe the pathological changes of liver and spleen tissues in mice. The expressions of phosphorylation signal transducer and activator of transcription (p-STAT3), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) and cysteine aspartate-specific protease-3 (Caspase-3) were detected in tumor tissues by Western blot. Result:In vitro, zeylenone had an obvious inhibitory effect on Molt-4 cells. IC50 values of zeylenone was 1.49 μmol·L-1. In vivo, compared with the model group, medium and high-dose zeylenone groups had significant tumor inhibition effects, with the inhibition rates of 50.24% and 60.75%, respectively (P<0.01). Additionally, liver and spleen injuries were slight in the above mentioned two groups compared with the vincristine group, indicating that zeylenone was safe. Western blot analysis showed that medium and high-dose zeylenone groups showed significant declines in proteins p-STAT3, Caspase-3 and Bcl-2, and marked increases in pro-apoptotic protein Bax compared with the model group (P<0.05, P<0.01). Conclusion:zeylenone could obviously inhibit the proliferation and induce the apoptosis of Molt-4 cells; and its mechanism may be related to the down-regulation of p-STAT3, Caspase-3, Bcl-2 and the up-regulation of Bax expressions. In addition, zeylenone had less damage to liver and spleen, and was safer than vincristine.