Molecular biological study of cytotoxicity due to leukotoxins and their metabolites

白细胞毒素及其代谢物细胞毒性的分子生物学研究

• 批准号: 12470135
• 负责人: ISHIZAKI Takeshi
• 金额: $ 7.17万
• 依托单位: Fukui Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470135/
• 关键词: Leukotoxin; Epoxide; Linoleate; Cell-cycle; Cyclin-dependent kinase; cyclin dependent kinase inhibitor; cell cycle; P16 INK4a; senescence; ディオール; cyclin dependent kinase; セルサイクル; Leukotoxin; Epoxide; Linoleate; Cell-cycle; Cyclin-dependent kinase; cyclin dependent kinase inhibitor; cell cycle; P16 INK4a; senescence; ディオール; cyclin dependent kinase; セルサイクル

We continued to study the effect of Leukotoxin (Lx), a linoleate epoxide, on the cell cycle. By using cultured human pulmonary arterial cells (HPEAC), we assessed the effect of antisense p16 INK4aS-oligo in terms of cell growth and used senescence-associated(SA)- β galactosidase staying for evaluating the degree of cell senescence in the presence/absence of Lx.Results : 1) Cell cycle of the HPEAC was arrested at the phase of G1 in the presence of Lx. The capability of DNA synthesis recovered when Lx was excluded or when pretreated with antisense p16INK4aS-oligo. The expression of mRNA and protein of p16INK4a was increased 3-fold at 48hr after addition of Lx and 4-fold at 96hr, respectively. However, either antisense p16 INK4aS-oligo or washout of Lx suppressed such an effect of Lx.2) When HPEAC passed PD50 the capability of growth of HPEAC was loosed and positive stained cells with SA-β galactosidase were increased. Whereas Lx suppressed the increase of SA-β galactosidase cells. The simultaneous presence of Lx and antisense p16INK4aS-oligo raised the cell number of positive SA-β galactosidase stain.Our current experiment suggests that Lx caused cell growth suppression and furthermore cell senescence suppression.

我们继续研究白细胞毒素（LX）（Linoleate环氧树脂）对细胞周期的影响。通过使用培养的人肺动脉细胞（HPEAC），我们在细胞生长方面评估了反义p16 ink4as-oligo的影响，并使用了与感应相关的（SA） - β半乳糖苷酶保持在LX.RES的存在/不存在HPEACECECECECECECECECECECECEAL CARPE CAILE CAILE CASE CARPE CAILE CASE AD AD the n a pEASTED AD the n d ar the n d ar the n d ar the n d ar the the the percecected n d rx的存在。当排除LX或用反义P16INK4AS-OLIGO预处理时，DNA合成的能力恢复了。在添加LX和96小时后，在48小时时，P16INK4A的mRNA和蛋白质的表达分别增加了3倍。然而，反义p16 ink4as-oligo或lx的冲洗都抑制了这种效果。2）当hpeac通过pd50时，HPEAC的生长能力松动，并增加了用SA-β半乳糖酶阳性染色的细胞。而LX抑制了SA-β半乳糖苷酶细胞的增加。 LX和反义P16INK4AS-OLIGO的简单存在增加了SA-β半乳糖苷酶染色的细胞数。