Effects of Paclitaxel and Quizartinib Alone and in Combination on AML Cell Line MV4-11 and Its STAT5 Signal Pathway / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of paclitaxel, quizartinib and their combination on proliferation, apoptosis and FLT3/STAT5 pathway of human leukemia cell line MV4-11 (FLT3-ITD+).@*METHODS@#MV4-11 cells were treated with paclitaxel and quizartinib at different concentrations for 24 h, 48 h and 72 h, respectively, and then the two drugs were combined at 48 h to compare the inhibition of proliferation, the apoptosis rate was detected by flow cytometry, the expression of FLT3 and STAT5 mRNA was determined by fluorescence quantitative PCR, and the protein expression of FLT3, p-FLT3, STAT5 and p-STAT5 was determined by Western blot.@*RESULTS@#Different combination groups of paclitaxel and quizartinib had synergistic inhibitory effect. The cell survival rate in the combination group was significantly lower than that in the single drug group (P<0.05). The cell apoptosis rate in the combination group was significantly higher than that in the single drug group (P<0.001). The expression of FLT3 mRNA in combination group was significantly higher than that in two single drugs (P<0.01). The expression of STAT5 mRNA in combination group was significantly higher than that in quizartinib group (P<0.001); increased compared with paclitaxel group, but there was no statistical significance. The expression level of p-FLT3、p-STAT5 protein in the combination group was significantly lower than that in the single drug group (P<0.05, P<0.05).@*CONCLUSION@#Paclitaxel combined with quizartinib can synergistically inhibit the proliferation of MV4-11 cell line and promote the apoptosis of MV4-11 cell line by inhibiting the activity of FLT3/STAT5 pathway.

Exploring the Mechanism of Paclitaxel Inhibiting T-cell Lymphoma based on High-throughput Sequencing and Public Databases / 中国实验血液学杂志

OBJECTIVE@#To analyze gene expression profile of T cell lymphoma Jurkat cell line treated with paclitaxel by computational biology based on next generation sequencing and to explore the possible molecular mechanism of paclitaxel resistance to T cell lymphoma at gene level.@*METHODS@#IC50 of paclitaxel on Jurkat cell line was determined by CCK-8 assay. Gene expression profile of Jurkat cells treated with paclitaxel was acquired by next generation sequencing technology. Gene microarray data related to human T cell lymphoma were screened from Gene Expression Omnibus (GEO) database (including 720 cases of T cell lymphoma and 153 cases of normal tissues). Combined with the sequencing data, differential expression genes (DEGs) were intersected and screened. DAVID database was used for enrichment analysis of GO function and KEGG pathway to determine and visualize functional entries of DEGs, and protein-protein interactions network of DEGs was drawn. The levels of gene expression were detected and verified by RT-qPCR.@*RESULTS@#CCK-8 results showed that the proliferation of Jurkat cells was inhibited by paclitaxel depended on the concentration apparently. Treated by paclitaxel for 48 h, P<0.05 and |log2(FC)|≥1 were used as filter criteria on the results of RNA Sequencing (RNA-Seq) and GeoChip, 351 DEGs were found from Jurkat cells, including 323 up-regulated genes and 28 down-regulated genes. The GO functional annotation and KEGG pathway enrichment analysis showed that the role of paclitaxel was mainly concentrated in protein heterodimerization activity, nucleosome assembly and transcriptional dysregulation in cancer, etc. The results of RT-qPCR were consistent with those of the sequencing analysis, which verified the reliability of this sequencing.@*CONCLUSION@#Paclitaxel can affect the proliferation and apoptosis of T-cell lymphoma by up-regulating JUN gene, orphan nuclear receptor NR4A family genes and histone family genes.

Effect of Mangiferin on Proliferation of FLT3-ITD Mutation Positive Acute Myeloid Leukemia Cell MV4-11 and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of mangiferin on proliferation, apoptosis and cycle of FLT3-ITD mutation-positive acute myeloid leukemia cells and its mechanism.@*METHODS@#The effects of different concentration of mangiferin on proliferation of MV4-11 cells were detected by CCK8 method. Apoptosis, cell cycle and FLT3 transmembrane protein expression were detected by flow cytometry. FLT3 mRNA expression was detected by real-time fluorescent quantitative polymerase chain reaction (PCR) .@*RESULTS@#Mangiferin obviously inhibited MV4-11 proliferation in a concentration and time dependent manner (48 h，r=0.922；72 h，r=0.959；96 h，r=0.973). The ratio of G0/G1 phase in cell cycle increased with the enhancement of concentration of mangiferin in MV4-11 cells for 48 h, and the ratio of S phase decreased with enhasment of concentration. The increase of apoptosis was more obvious. The expression of FLT3 transmembrane protein significantly decreased after the actior of IC50 concentration of mangiferin in MV4-11 cells for 48 h. The results of qRT-PCR showed that the expression of FLT3 mRNA significantly decreased after treatment of MN4-11 cells with mangiferin (P＜0.05).@*CONCLUSION@#Mangiferin inhibits MV4-11 cell proliferation, arrests cell cycle progression and promotes apoptosis, which may be related with the inhibition of FLT3 activity by mangiferin and the subsequent signaling pathways involved in apoptosis and proliferation of cells.

Changes of serum anti-schistosome antibody levels in schistosomiasis japoni-ca patients after treatment / 中国血吸虫病防治杂志

Objective To investigate the changes of serum anti-schistosome antibody titers in schistosomiasis japonica pa-tients after treatment,in order to provide the evidence for formulating the schistosomiasis surveillance program in marshland and lake regions.Methods Upon prospective cohort study,the stool examination positive schistosomiasis patients and blood exami-nation positive suspected patients(the titer was more than 1:80,including 1:80)were selected as the research objects in Jian-gling County in 2014,and they received the 2-day praziquantel therapy.Half year,one year and two years after the treatment, their blood samples and fecal samples were collected for IHA anti-schistosome antibody detections and schistosome egg and mira-cidium detections. Results In 2014,the stool examination positives were 251,and the majority of them were over 41 years old,accounting for 93.23%(234/251);581 cases of high antibody titers were detected by the IHA method,and the majority of them were over 41 years old,accounting for 89.16%(518/581).Half year,one year and two years after the treatment,among the stool examination positives,the negative conversion rates of stool positives were 99.60%(250/251),100%(239/239)and 100%(234/234)respectively and the negative conversion rates of antibody positives were 21.91%(55/251),64.11%(156/239)and 76.89%(193/234)respectively.In the high antibody titer positives,the negative conversion rates were 38.04%(221/581),64.11%(359/560),and 77.86%(429/551)respectively,Half year,one year and two years after the treatment.There were statistically significant differences among the antibody negative conversion rates by χ2test(χ2=77.538,183.412,25.469 respectively,all P<0.001).The geometric mean values of antibody titers of different durations between 2 groups were analyzed by 2-independent-samples T test,and the geometric mean values of antibody titers between the 2 groups were different before the treatment(t=23.576,P<0.01),but the geometric mean values of antibody titers between the 2 groups were not different 6 months,1 year and 2 years after the treatment(t=-0.046,1.165, -0.132,P=0.964, 0.245,0.895 respectively). Conclu-sions The levels of serum anti-schistosome antibody degrade slowly in schistosomiasis japonica patients after the treatment, and the results of IHA tests cannot distinguish the current schistosome infection from previous schistosome infection.Therefore, it is necessary to develop the specific diagnostic technology for schistosome infection in order to meet the need of monitoring.