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甾醇类药NSC67657诱导急性早幼粒细胞白血病HL-60细胞单核系分化中Wnt信号通路相关蛋白的表达 被引量:3

Expression of ICAT and Wnt signaling-related proteins in the monocytic differentiation of HL-60 cells induced by a new steroidal drug NSC67657
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摘要 目的:探讨甾醇类药NSC67657诱导急性早幼粒细胞白血病HL-60细胞单核系分化时β连环蛋白(β-catenin)和T细胞因子4( TCF-4)抑制蛋白( ICAT)和Wnt信号通路相关mRNA和蛋白的表达。方法10μmol/L NSC67657药物处理HL-60细胞5 d后,采用瑞氏染色、非特异酯酶染色分析细胞的分化方向,采用流式细胞术分析细胞的分化程度和细胞周期的分布情况,采用逆转录聚合酶链反应和Western blot检测 Wnt 信号通路相关因子 ICAT、β-catenin、TCF-4、Wnt 信号通路下游靶点c-myc、细胞周期蛋白D1( cyclin D1)、TCF-1 mRNA和蛋白在细胞分化前后的表达情况。结果 NSC67657可诱导HL-60细胞向单核系分化。10μmol/L NSC67657作用5 d后,处理组CD14+细胞数为(94.37±2.84)%,高于未处理组[(1.31±0.09)%],差异有统计学意义(P<0.01)。处理组G1/G0期细胞所占比例为(76.46±2.83)%,高于未处理组[(59.40±5.42)%],差异有统计学意义(P<0.05);处理组S期细胞所占比例为(18.76±0.98)%,低于未处理组[(34.38±2.61)%],差异有统计学意义(P<0.05)。药物作用后HL-60细胞中ICAT mRNA和蛋白的表达水平上调(均P<0.01),β-catenin mRNA和蛋白的表达水平下调(均P<0.01),细胞核中β-catenin蛋白的表达水平下调( P<0.01)。处理组和未处理组HL-60细胞中TCF-4 mRNA、总蛋白和细胞核蛋白的表达水平差异均无统计学意义(均P>0.05)。药物诱导HL-60细胞分化后,Wnt信号通路下游靶基因c-myc、cyclin D1、TCF-1 mRNA和蛋白的表达水平均下调(均P<0.05)。结论 NSC67657可诱导HL-60细胞向单核系分化,导致Wnt信号通路关键调节因子β-catenin及下游靶点的表达下调,Wnt信号通路可能直接或间接地参与了HL-60细胞的单核系分化进程。 Objective To investigate the expression of mRNA and proteins of β-catenin, TCF-4 ( ICAT) and Wnt signaling pathway-related genes in the monocytic differentiation of acute myeloid leukemia HL-60 cells induced by a new steroidal drug NSC67657. Methods Wright′s staining and α-NBE staining were used to observe the differentiation of HL-60 cells after 5 days of 10μmol/L NSC67657 treatment. Flow cytometry ( FCM ) was used to detect the differentiation and cell cycles. The expressions of mRNA and proteins of ICAT and Wnt signaling pathway-related factors, including β-catenin, TCF-4, c-myc, cyclin D1 and TCF-1 before and after differentiation, were detected by RT-PCR and Western blot. Results Morphological observation showed that NSC67657 induced monocytic differentiation of HL-60 cells. At 5 days after 10 μmol/L NSC67657 treatment, the number of CD14+ HL-60 cells was (94.37±2.84)%, significantly higher than the (1.31±0.09)% in control group (P〈0.01). The flow cytometry assay revealed that NSC67657 induced (76.46±2.83)% of G1/G0 phase arrest, significantly higher than that of (59.40±5.42)% in the control group (P〈0.05), while the S phase cells were of (18.76±0.98)%, significantly lower than that of (34.38±2.61) % in the control group (P〈0.05). The NSC67657 treatment also up-regulated the expression of ICAT mRNA and protein, and down-regulated the expression ofβ-catenin mRNA and protin ( P〈0.01 for all) . However, the nuclear expression ofβ-catenin was down-regulated ( P〈0.01) . The NSC67657 treatment induced nonsignificant alterations of TCF-4 mRNA, total protein and nuclear protein in the HL-60 cells ( P〉0.05 for all) . The target genes of Wnt signaling pathway, including c-myc, cyclinD1 and TCF-1 mRNA and proteins in the HL-60 cells were significantly down-regulated after NSC67657 treatment ( P〈0. 05 ) . Conclusions The new steroidal drug NSC67657 induces monocytic differentiation of HL-60 cells, and down-regulates the expression
作者 王晋蜀 王伟佳 王婷 张彦 Wang Jinshu, Wang Weijia, Wang Ting, Zhang Yan(1. Key Laboratory of Clinical Laboratory Diagnostics of Ministry of Education, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China; 2.Zhongshan People's Hospital Medical Laboratory Center, Zhongshan, Guangdong 528403, China)
出处 《中华肿瘤杂志》 CAS CSCD 北大核心 2016年第4期246-251,共6页 Chinese Journal of Oncology
基金 国家自然科学基金(81301492)
关键词 白血病 早幼粒细胞 急性 WNT蛋白质类 HL-60细胞 细胞分化 NSC67657 β连环蛋白和T细胞因子4抑制蛋白 Leukemia, promyelocytic, acute Wnt proteins HL-60 cells Monocytic differentiation NSC67657 Inhibitor of β-catenin and TCF-4
作者简介 通信作者:张彦,Email:zy2753@hotmail.com
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