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Pressure-induced cell death in human immunocytes.

压力诱导的人类免疫细胞细胞死亡。

基本信息

批准号：
09672315
负责人：
TAKANO Takako
金额：
$ 1.98万
依托单位：
Teikyo University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672315/
关键词：
pressure cell death apoptosis lymphoblast Ramos HL-60 immunocyte

项目摘要

As pressure is the fundamental physical quantity, the knowledge of the role of pressure is useful to understand the whole scheme of cell death. The viability of human immunocytes was investigated after compression at 37ﾟC for 30 minutes using a high pressure apparatus of the piston-cylinder type. In the case of EB3 (EB virus transformed human lymphoblasts), necrotic cells were observed immediately after decompression, and the viability decreased sigmoidally to zero with increasing pressure from 85 to 250 MPa. A 50% cell viability (P_<1/>) was attained at a pressure of 170 MPL Apoptotic cells began to be observed from 8 knirs after decompression (compression at 100 MPa for 30 minutes at 37ﾟC). The apoptosis was confirmed to be pressure-induced by the TUNEL (TdT-mediated dUll' nick end labeling) technique and by optical and electron microscopy. In the case of 50 MPa, the cell counts increased gradually after compression, which means that cell division progressed.Qualitatively similar res … More ults were obtained with B cell lymphoma (Ramos). P_<1/> in HL-60 (human promyelocytic leukemia cells) and Ramos was lower than that in EB3. In the case of HL-60, apoptotic cells were observed immediately after decompression (compression at 100 MPa for 30 minutes at 37ﾟC). Execution of apoptosis should be different between EB3 and HL-60. Expression of protein related to pressure-induced apoptosis was studied using Western blot analysis. In EB3 cells, compared with uncompressed control, the expression of p53 and Bax increased from 5 lxurs after compression at IOOMPa, whereas that of caspase-3 (CPP32, apoptotic cysteine protease) decreased. Overexpression of p53 may induce caspase-3 processing through Bax activation. The pressure-induced attenuation in a-tubulin expression was observed after compression at lOOMPa, while it was not observed at 50 MPa. HL-60 is p53 deleted cell line. In LIL-60 cells, the expression of phospho p38 MAPK (mitogen-activated protein kinase), Bcl-2 and Bax increased immediatly after decompression at lOOMPa After 2 hours, the expression of an inactive 32-kDa precusor of caspase-3 (CPP32) decreased, and the mature activated p17 form of caspase-3 was detected. These findings suggest that p38 MAPK cascade may be induced by high-pressure, and the participation in the common signaling pathway (caspase cascade) may lead to apoptosis in HL-60 cells.Our study of human cell biology at high pressure aids the medical application of high pressure in the near future. Less

由于压力是基本的物理量，了解压力的作用有助于理解细胞死亡的整个过程。使用活塞-气缸型高压装置在37 ℃下压缩30分钟后研究人免疫细胞的存活力。在EB 3（EB病毒转化的人淋巴母细胞）的情况下，减压后立即观察到坏死细胞，并且随着压力从85 MPa增加到250 MPa，存活率呈S形下降到零。在170 MPL的压力下达到50%的细胞存活率（P_）。在减压（在37 ℃、100 MPa下压缩30分钟）后，从8小时开始观察到凋亡细胞。用TUNEL（TdT-mediated dUll' nick end labeling）技术及光镜、电镜观察证实细胞凋亡是压力诱导的。在50 MPa的情况下，细胞计数在压缩后逐渐增加，这意味着细胞分裂进行。 ...更多信息 B细胞淋巴瘤（拉莫斯）获得结果。HL-60细胞和拉莫斯细胞的P_（max）值低于EB_3细胞。在HL-60的情况下，减压后立即观察到凋亡细胞（在37 ℃下以100 MPa压缩30分钟）。EB 3和HL-60细胞凋亡的实施应该是不同的。Western blot分析压力诱导细胞凋亡相关蛋白的表达。在EB 3细胞中，与未压缩的对照相比，在100 MPa压缩后，p53和Bax的表达从5个小时开始增加，而caspase-3（CPP 32，凋亡半胱氨酸蛋白酶）的表达下降。p53的过表达可能通过Bax的激活诱导caspase-3的加工。在100 MPa下压缩后观察到α-微管蛋白表达的压力诱导的衰减，而在50 MPa下未观察到。HL-60是p53缺失的细胞系。LIL-60细胞在100 MPa压力下减压后，磷酸化p38 MAPK（mitogen-activated protein kinase）、Bcl-2和Bax的表达立即增加。减压2小时后，失活的32 kDa caspase-3前体（CPP 32）的表达减少，并检测到成熟的激活的p17型caspase-3。这些结果提示高压可诱导p38 MAPK级联反应，参与共同信号通路（caspase级联反应）可能导致HL-60细胞凋亡。少