sincalide and Leukemia--Myeloid--Acute

Mitosis-associated genes are dysregulated in many types of cancers and play important roles in disease progression and chemotherapy resistance. However, their expression and functions in chemotherapy-resistant Acute Myeloid Leukemia (AML) are still largely undetermined.. This study aims to explore the roles of spindle assembly checkpoint (SAC) genes CENPE, CENPF, and DLGAP5 in chemotherapy-resistant AML.. RNA-sequencing (RNA-seq) was performed in patients with chemotherapy-resistant AML and chemotherapy-sensitive AML. AML mRNA data from 151 patients with recurrence were downloaded from TCGA. Integrated analysis of the differentially expressed genes (DEGs), GO and KEGG pathways. CENPE, CENPF, or DLGAP5 knockdown cell lines were used to analyse proliferation, apoptosis and cell cycle alterations.. A total of 87 DEGs (48 upregulated and 39 downregulated) were obtained through gene analysis of R/R-AML and a total of 329 DEGs (202 upregulated and 127 downregulated) were obtained in refractory S-AML. Upregulated DEGs were mainly enriched in cell cycle (GO: 0007049, hsa04110) and mitotic cell cycle (GO: 0000278) processes and pathway. Venn diagram analysis identified the most upregulated DEGs (including CENPE, CENPF, and DLGAP5) in chemoresistant AML. The expression of CENPE, CENPF and DLGAP5 in R-AML (TCGA) was significantly higher than that of primary AML (GEO). The proliferation of K562 cells after CENPE and DLGAP5 knockdown was significantly decreased (P= 0.0001 and P= 0.0006). In THP-1 cells, the CCK-8 values after CENPE, CENPF and DLGAP5 knockdown were significantly decreased (P= 0.01, P= 0.0395 and P= 0.0362). Knockdown of CENPE, CENPF and DLGAP5 significantly increased cell apoptosis by regulating Caspase-9, BAX, TP-53 and bcl-2, and induced cell cycle arrested by regulating CDK1, CDK2, CDKN1A, and CyclinD1.. In conclusion, the mitotic cell cycle-associated genes CENPE, CENPF, and DLGAP5 were upregulated in chemotherapy-resistant AML patients and might be useful for predicting poor prognosis.

Topics: bcl-2-Associated X Protein; Caspase 9; Humans; Leukemia, Myeloid, Acute; Mitosis; Neoplasm Proteins; Prognosis; RNA, Messenger; Sincalide

To investigate the expression of ROBO3 in pediatric AML patients and explore its function on cell proliferation and apoptosis.. The expression of ROBO3 in pediatric AML patients at different treatment stage was detected by real-time quantitative polymerase chain reaction (RT-qPCR). The relationship between the expression of ROBO3 and clinic pathological characteristics in newly diagnosed pediatric AML patients was analyzed. Moreover, the effects of ROBO3 on the proliferation and apoptosis of AML cell lines HL-60 and THP-1 were estimated by using CCK-8 and flow cytometry after transfection with ROBO3 siRNA.. It was found that ROBO3 expression was significantly increased in most of newly diagnosed pediatric AML patients, especially in non-M3 subtype, younger patients (<10 years old), and high risk group, compared to corresponding controls. Furthermore, the expression level of ROBO3 was sharply decreased in patients who achieved complete remission. Targeting ROBO3 significantly inhibited AML cell proliferation, as well as increased apoptosis by ROBO3 siRNA transfection in vitro.. ROBO3 is differentially expressed within distinct subtypes of the pediatric AML patients, which suggested that ROBO3 may be a potential biomarker and a new therapeutic target for pediatric AML.. ROBO3在儿童急性髓系白血病患者中的表达及其对细胞增殖和凋亡的影响.. 研究ROBO3在儿童急性髓系白血病（AML）患者中的表达及其对AML细胞增殖和凋亡的影响。.. 采用荧光定量PCR法检测不同治疗阶段的儿童AML患者骨髓中ROBO3的表达，分析初诊患儿ROBO3表达量与临床特征的关系。同时采用电转染的方法靶向干扰ROBO3表达，应用 CCK-8 法检测其对AML细胞系（HL-60和THP-1）增殖的影响，流式细胞术检测其对凋亡的作用。.. 与健康对照组相比，ROBO3在初诊儿童AML患者中显著高表达，尤其是在非M3型、年龄偏小的患者（<10岁）及高危组中高表达。而且与初发组相比，治疗后完全缓解组ROBO3 mRNA的表达水平显著下降。靶向ROBO3可以有效抑制AML细胞的增殖，并促进细胞凋亡。.. ROBO3在儿童AML患者中差异表达，很可能是儿童AML潜在的标志物及新靶点。.

Topics: Apoptosis; Cell Line, Tumor; Cell Proliferation; Child; Humans; Leukemia, Myeloid, Acute; MicroRNAs; Receptors, Cell Surface; RNA, Small Interfering; Sincalide

Microrna-143 (miR-143) has been suggested to be a tumor suppressor, yet its role in hematological tumors has not been determined. Thus, we aimed to explore the expression and function of miR-143 in leukemia cells. miR-143 expression was assessed in bone marrow samples from 63 leukemia patients and 15 healthy controls using q-PCR, and its correlation with DNMT3A expression was determined. In addition, after lentiviral-mediated miR-143 overexpression, K562 cell proliferation was evaluated using CCK-8 analysis; cell cycle progression and apoptosis were determined using flow cytometry. The expression of Bcl-2 and pro-caspase-3 and -9 was assessed by q-PCR and western blot analysis, respectively. Leukemia patients had significantly lower relative miR-143 expression than healthy controls (P=0.004), and the expression levels of miR143 and DNMTA3A were negatively correlated (r=-0.663, P=0.001). Overexpression of miR-143 decreased DNMT3A mRNA and protein expression, and significantly reduced K562 cell proliferation at 72 and 96 h (both P ≤ 0.018). In addition, reduced colony formation and cell cycle progression were observed upon miR-143 overexpression. Flow cytometric analysis revealed that the early apoptosis rate was higher in the miR-143 group than the rate in the NC group. Bcl-2 mRNA expression and pro-caspase-3 and -9 protein expression were reduced in the miR-143-expressing cells. These findings suggest that miR-143 plays an important role in leukemia cell proliferation and apoptosis, possibly through silencing of DNMT3A. Further studies are necessary to determine the prognostic value and therapeutic potential of targeting miR-143.

Topics: Adolescent; Adult; Aged; Apoptosis; Bone Marrow Cells; Caspase 3; Caspase 9; Cell Line, Tumor; Cell Proliferation; Child; DNA (Cytosine-5-)-Methyltransferases; DNA Methyltransferase 3A; Female; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; Humans; Leukemia, Myeloid, Acute; Male; MicroRNAs; Middle Aged; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Proto-Oncogene Proteins c-bcl-2; RNA, Messenger; Sincalide; Young Adult