Regulatory mechanism between autophagy and apoptosis in leukemia cells

白血病细胞自噬与凋亡的调控机制

• 批准号: 18591089
• 负责人: MIYAZAWA Keisuke
• 金额: $ 1.71万
• 依托单位: Tokyo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591089/
• 关键词: autophagy; apoptosis; leukemia; bcl-2; cell death; vitamin K2

We investigated molecular based regulatory mechanism between autophagy and apoptosis.Vitamin K2 (menaquinone-2 : VK2) is now known to be a potent inducer for apoptosis in leukemia cells in vitro. HL-60bc1-2 cells, which are derived from a stable transfectant clone of human bc1-2 gene into HL-60 leukemia cell line, show 5-fold greater expression of Bc1-2 protein compared with that in HL-60neo cells, a control clone transfected with vector alone. Although HL-60neo cells are induced apoptosis in response to VK2, HL-60bc1.2 cells are resistant against apoptosis induction but still show cell growth inhibition along with an increase of cytoplasmic vacuoles during exposure to VK2. Electron microscopy revealed autophagosomes and autolysosomes formation in HL-60bc1-2 cells after exposure to VK2. An increase of acid vesicular organelles (AVO) detected by acridine orange staining for lysosomes as well as conversion of LC3B-I into LC3B-II by immunonoblotting and an increased punctuated pattern of cytoplasmic LC3B by fluorescent immunostaining all supported enhanced autophagy induction in response to VK2 in HL-60bc1-2 cells. However, during shorter exposure to VK2, autophagosome formation was rather prominent in HL-60neo cells although nuclear chromatin condensations and nuclear fragments were also observed at the same time. These findings indicated the mixed morphologic features of apoptosis and autophagy. Inhibition of autophagy by either addition of 3-methyladenine, siRNA for Atg7, or Tet-off AtgS system all resulted in attenuation of VK2-incuded cell death, indicating autophagy-mediated cell death in response to VK2. These data demonstrate that autophagy and apoptosis can be simultaneously induced by VK2. However, autophagy becomes prominent when the cells were protected from rapid apoptotic death by higher expression level of Bcl-2.

维生素K2（menaquinone-2：VK 2）是一种有效的诱导白血病细胞凋亡的因子，在体外研究了自噬和凋亡之间的分子调控机制。将人bcl-2基因稳定转染HL-60白血病细胞系，获得HL-60 bc 1 -2细胞，其bcl-2蛋白表达量是对照细胞HL-60 neo的5倍。尽管HL-60 neo细胞在VK 2作用下可被诱导凋亡，但HL-60 bc1.2细胞对凋亡诱导具有抵抗性，但在暴露于VK 2期间仍显示细胞生长抑制沿着细胞质空泡的增加。电子显微镜观察显示，HL-60 bc 1 -2细胞暴露于VK 2后，自噬体和自溶酶体形成。通过吖啶橙子染色检测到的溶酶体酸性囊泡细胞器（AVO）的增加，以及通过免疫印迹检测到的LC 3B-I转化为LC 3B-II，以及通过荧光免疫染色检测到的胞质LC 3B的点状图案的增加，这些都支持HL-60 bc 1 -2细胞对VK 2的应答增强的自噬诱导。然而，在较短的暴露于VK 2，自噬体的形成是相当突出的HL-60 neo细胞，虽然核染色质浓缩和核碎片也在同一时间观察到。这些结果表明，细胞凋亡和自噬的混合形态特征。通过添加3-甲基腺嘌呤、针对Atg 7的siRNA或Tet-off AtgS系统抑制自噬均导致VK 2引起的细胞死亡的减弱，表明自噬介导的细胞死亡响应于VK 2。这些数据表明，自噬和凋亡可以同时诱导VK 2。然而，自噬变得突出时，细胞受到保护，从快速凋亡死亡的Bcl-2的表达水平较高。

项目成果

VitaminK2は胆管癌細胞株TFK-1に対してautophagyを伴った抗腫瘍効果を有する。

维生素K2 对胆管癌细胞系 TFK-1 具有抗肿瘤作用，并伴有自噬作用。

• 发表时间: 2007
• 作者: 榎本 正統;土田 明彦;宮澤 啓介;横山 智央;鴇田 博美;内藤 宗和;伊藤 正裕;大屋敷 一馬;青木 達哉
• 通讯作者: 青木 達哉

Clinical features of adult leukemia with 11q23 abnormalities in Japan : a co-operative multicenter study

日本 11q23 异常成人白血病的临床特征：一项合作多中心研究

• 发表时间: 2007
• 期刊: Int J Hematol 87
• 作者: Tamai;H.;Yamaguchi;H.;Hamaguchi;H.;Miyazawa;K.;et. al.
• 通讯作者: et. al.

Clinical features of hypereosinophilic syndrome : FIPILI-PDGFRA fusion gene-positive disease is a distinct clinical entity with myeloproliferative features and a poor response to corticosteroid.

嗜酸性粒细胞增多综合征的临床特征：FIPILI-PDGFRA 融合基因阳性疾病是一种独特的临床实体，具有骨髓增殖特征且对皮质类固醇反应不佳。

• 发表时间: 2007
• 期刊: Int J Hematol. 85・1
• 作者: Miyazawa K;Kakazu N;Ohyashiki K
• 通讯作者: Ohyashiki K

Translocation(10;21)(q21;q22)in myelodysplastic syndrome.

骨髓增生异常综合征中的易位(10;21)(q21;q22)。

• 发表时间: 2007
• 期刊: Cancer Genet Cytogenet. 173・1
• 作者: Rashaan Z;Ito Y;Takaku T;Toyotake J;Miyazawa K;et al.
• 通讯作者: et al.

Vitamin K2-induced cell growth inhibition via autophagy formation in cholangiocellular carcinoma cell lines.

• DOI: 10.3892/ijmm.20.6.801
• 发表时间: 2007-12
• 期刊: International journal of molecular medicine
• 影响因子: 5.4
• 作者: M. Enomoto;A. Tsuchida;K. Miyazawa;Tomohisa Yokoyama;Hideaki Kawakita;H. Tokita;M. Naito;M. Itoh;K. Ohyashiki;T. Aoki