Molecular mechanism of G(1) arrest and cellular senescence induced by LEE011, a novel CDK4/CDK6 inhibitor, in leukemia cells.
Molecular mechanism of G(1) arrest and cellular senescence induced by LEE011, a novel CDK4/CDK6 inhibitor, in leukemia cells.
复制标题
新型CDK4/CDK6抑制剂LEE011诱导白血病细胞G(1)阻滞和细胞衰老的分子机制
DOI：
10.1186/s12935-017-0405-y
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发表时间：
2017
影响因子：
5.8
通讯作者：
Pan J
中科院分区：
文献类型：
--
作者：
Tao YF;Wang NN;Xu LX;Li ZH;Li XL;Xu YY;Fang F;Li M;Qian GH;Li YH;Li YP;Wu Y;Ren JL;Du WW;Lu J;Feng X;Wang J;He WQ;Hu SY;Pan J
细胞周期蛋白D1依赖性激酶4和6（CDK 4/6）的过表达是包括白血病在内的许多人类癌症的共同特征。LEE 011是CDK 4和CDK 6的新型抑制剂。迄今为止，LEE 011在白血病中的分子功能仍不清楚。通过CCK-8和膜联蛋白V/碘化丙啶染色测定评估LEE 011处理后的白血病细胞生长和凋亡。通过β-半乳糖苷酶染色和p16 INK 4a表达分析来评估细胞衰老。使用Arraystar Human LncRNA阵列研究LEE 011处理的HL-60细胞的基因表达谱。然后利用基因本体论和KEGG通路分析对聚类分析得到的差异表达基因进行分析。我们的研究表明LEE 011能抑制白血病细胞增殖并诱导细胞凋亡。Hoechst 33，342染色分析显示LEE 011处理后DNA断裂和核结构变形。细胞周期分析显示，LEE 011在8个急性白血病细胞系中的7个中显著诱导细胞周期G1停滞，THP-1细胞除外。β-半乳糖苷酶染色分析和p16 INK 4a表达分析显示LEE 011处理可诱导白血病细胞衰老。LncRNA微阵列分析显示，与对照组相比，LEE 011处理的HL-60细胞中差异表达的mRNA为2083个，差异表达的lncRNA为3224个。分子功能分析表明，LEE 011诱导白血病细胞衰老部分是通过下调MYBL 2的转录表达。我们首次证明LEE 011处理导致白血病细胞的细胞增殖抑制和G1期阻滞和细胞衰老的诱导。LncRNA微阵列分析显示LEE 011处理的HL-60细胞中差异表达的mRNA和lncRNA，我们证明LEE 011部分通过下调MYBL 2的表达诱导细胞衰老。这些结果可能为LEE 011诱导细胞衰老的分子机制开辟新的研究方向。本文的在线版本（doi：10.1186/s12935-017-0405-y）包含补充材料，可供授权用户使用。
Overexpression of cyclin D1 dependent kinases 4 and 6 (CDK4/6) is a common feature of many human cancers including leukemia. LEE011 is a novel inhibitor of both CDK4 and 6. To date, the molecular function of LEE011 in leukemia remains unclear. Leukemia cell growth and apoptosis following LEE011 treatment was assessed through CCK-8 and annexin V/propidium iodide staining assays. Cell senescence was assessed by β-galactosidase staining and p16INK4a expression analysis. Gene expression profiles of LEE011 treated HL-60 cells were investigated using an Arraystar Human LncRNA array. Gene ontology and KEGG pathway analysis were then used to analyze the differentially expressed genes from the cluster analysis. Our studies demonstrated that LEE011 inhibited proliferation of leukemia cells and could induce apoptosis. Hoechst 33,342 staining analysis showed DNA fragmentation and distortion of nuclear structures following LEE011 treatment. Cell cycle analysis showed LEE011 significantly induced cell cycle G1 arrest in seven of eight acute leukemia cells lines, the exception being THP-1 cells. β-Galactosidase staining analysis and p16INK4a expression analysis showed that LEE011 treatment can induce cell senescence of leukemia cells. LncRNA microarray analysis showed 2083 differentially expressed mRNAs and 3224 differentially expressed lncRNAs in LEE011-treated HL-60 cells compared with controls. Molecular function analysis showed that LEE011 induced senescence in leukemia cells partially through downregulation of the transcriptional expression of MYBL2. We demonstrate for the first time that LEE011 treatment results in inhibition of cell proliferation and induction of G1 arrest and cellular senescence in leukemia cells. LncRNA microarray analysis showed differentially expressed mRNAs and lncRNAs in LEE011-treated HL-60 cells and we demonstrated that LEE011 induces cellular senescence partially through downregulation of the expression of MYBL2. These results may open new lines of investigation regarding the molecular mechanism of LEE011 induced cellular senescence. The online version of this article (doi:10.1186/s12935-017-0405-y) contains supplementary material, which is available to authorized users.