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The elucidation of the apoptosis inhibitory action of SMP30.

阐明SMP30的细胞凋亡抑制作用。

基本信息

批准号：
15603010
负责人：
HANDA Setsuko
金额：
$ 2.3万
依托单位：
Tokyo Metropolitan Foundation for Research on Aging and Promotion of Human Welfare
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15603010/
关键词：
apoptosis SMP30 aging liver Akt knockout mice vitamin C gluconolactonase TNF-α アクチノマイシンD

项目摘要

Senescence marker protein-30 (SMP30) is a 34-kDa protein whose tissue levels in the liver, kidney, and lung decrease with aging. To elucidate the physiological role of this protein, we introduced a null mutation of the SMP30 gene into the germ line of mice, and then investigated the tissue susceptibility for apoptosis induced by tumor necrosis factor-alpha (TNF-alpha) using primary cultured hepatocytes. Consequently, cells that are completely lacking SMP30 (SMP30-/-) were more susceptible to apoptosis induced by TNF-alpha plus actinomycin D (Act-D) than SMP30+/+ hepatocytes, indicating that SMP30 can protect cells from apoptosis induced by TNF-alpha plus Act-D. However, the responsible mechanism(s) for anti-apoptotic effect of SMP30 has not been fully understood. Therefore we aimed in this study that the elucidation of the apoptosis inhibitory action of SMP30.Human hepatocellular carcinoma cell line was transfected with pcDNA3/SMP30 (SMP30 transfectants), or as a control with pcDNA3 (mock transfectants). When cells were exposed to 20 ng/ml tumor necrosis factor-alpha (TNF-alpha) plus 10 ng/ml actinomycin D (Act-D) for 15 h, the viability of cells was decreased in both SMP30 and mock transfectants. However, the viability of cells was threefold higher in SMP30 transfectants than mock transfectants. Cell death was confirmed as apoptosis by TUNEL assay. The presence of trifluoperazine, a calmodulin (CaM) inhibitor, attenuated anti-apoptotic effect of SMP30 in both transfectants, but the effect was more prominent in SMP30 transfectants. Western blot analyses revealed that Akt, which acts as a survival factor in cells, was activated in SMP30, but not mock, transfectants either in the presence or absence of TNF-alpha plus Act-D. Further, trifluoperazine inhibited Akt activation in SMP30 transfectants. We therefore propose that interplay between CaM and SMP30 regulates Akt activity, and thus SMP30 acts as a survival factor in hepatocytes.

衰老标记蛋白 30 (SMP30) 是一种 34 kDa 的蛋白质，其在肝脏、肾脏和肺中的组织水平随着衰老而降低。为了阐明该蛋白的生理作用，我们将 SMP30 基因的无效突变引入小鼠种系，然后使用原代培养的肝细胞研究肿瘤坏死因子-α (TNF-α) 诱导的细胞凋亡的组织敏感性。因此，完全缺乏SMP30的细胞（SMP30-/-）比SMP30+/+肝细胞更容易受到TNF-α加放线菌素D（Act-D）诱导的细胞凋亡的影响，表明SMP30可以保护细胞免受TNF-α加Act-D诱导的细胞凋亡。然而，SMP30 抗凋亡作用的机制尚未完全清楚。因此，我们本研究的目的是阐明SMP30的细胞凋亡抑制作用。用pcDNA3/SMP30（SMP30转染子）转染人肝癌细胞系，或用pcDNA3（模拟转染子）作为对照。当细胞暴露于 20 ng/ml 肿瘤坏死因子-α (TNF-α) 加 10 ng/ml 放线菌素 D (Act-D) 15 小时时，SMP30 和模拟转染子中的细胞活力均下降。然而，SMP30 转染子中的细胞活力比模拟转染子高出三倍。通过TUNEL测定确认细胞死亡为细胞凋亡。钙调蛋白 (CaM) 抑制剂三氟拉嗪的存在减弱了两种转染子中 SMP30 的抗凋亡作用，但该作用在 SMP30 转染子中更为突出。蛋白质印迹分析显示，在存在或不存在 TNF-α 加 Act-D 的情况下，作为细胞存活因子的 Akt 在 SMP30（而非模拟转染子）中被激活。此外，三氟拉嗪抑制 SMP30 转染子中的 Akt 激活。因此，我们认为 CaM 和 SMP30 之间的相互作用调节 Akt 活性，因此 SMP30 充当肝细胞中的生存因子。

项目成果

期刊论文数量（48）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

SMP30 Functions as Gluconolactonase in L_Ascorbic Acid Biosynthesis and Its Knockout Mice Are Prone to Scurvy.

SMP30在L_抗坏血酸生物合成中充当葡萄糖酸内酯酶，其基因敲除小鼠易患坏血病。

DOI：
发表时间：
2006
期刊：
Proc. Nat. Acad. Sci. USA in press
影响因子：
0
作者：
Jung et al.;Son et al.;Kondo et al.
通讯作者：
Kondo et al.

SMP30 Functions as Gluconolactonase in L-Ascorbic Acid Biosynthesis and Its Knockout Mice Are Prone to Scurvy.

SMP30 在 L-抗坏血酸生物合成中充当葡萄糖酸内酯酶，其基因敲除小鼠容易患坏血病。

DOI：
发表时间：
2006
期刊：
Proc.Nat.Acad.Sci.USA (in press)
影响因子：
0
作者：
Jung et al.;Son et al.;Kondo et al.;Jung et al.;Son et al.;Kondo et al.
通讯作者：
Kondo et al.

Senescence Marker Protein-30 (SMP30) induces formation of microvilli and bile canaloculi in hep G2 cells.

衰老标记蛋白 30 (SMP30) 诱导 Hep G2 细胞中微绒毛和胆小管的形成。

DOI：
发表时间：
2005
期刊：
Cell & Tissue Res. 320
影响因子：
0
作者：
Jung et al.;Son et al.;Kondo et al.;Jung et al.;Son et al.;Kondo et al.;Ishigami et al.
通讯作者：
Ishigami et al.

Accelerated tubular cell senescence in SMP30 knockout mice.

DOI：
10.14670/hh-21.1151
发表时间：
2006-11
期刊：
Histology and histopathology
影响因子：
2
作者：
W. Yumura;T. Imasawa;S. Suganuma;A. Ishigami;S. Handa;S. Kubo;K. Joh;N. Maruyama
通讯作者：
W. Yumura;T. Imasawa;S. Suganuma;A. Ishigami;S. Handa;S. Kubo;K. Joh;N. Maruyama

Modulation of gene expression of SMP-30 by LPS and calorie restriction during aging process