CAJ | 학술논문

目的探讨CD45分子的表达与骨髓瘤细胞凋亡的关系.方法用马法兰诱导骨髓瘤细胞系U266的凋亡;无血清培养法诱导转染CD45基因或空质粒的U266细胞凋亡;无葡萄糖培养法诱导AMO1细胞凋亡;动物实验比较U266细胞CD45阴性和阳性两群细胞在体内的存活能力;流式细胞术检测凋亡细胞率和线粒体膜电位;Western blot法检测线粒体内细胞色素C的释放和caspase-9的激活.结果经马法兰处理后,45%的CD45+U266细胞发生凋亡,而CD45-U266细胞只有30%发生凋亡;在无葡萄糖培养条件下,与低表达CD45的AMO1细胞相比,高表达CD45的AMO1细胞更易发生凋亡.无血清培养5 d,转染CD45RB基因的U266细胞60%发生凋亡,而转染空质粒的U266细胞的凋亡数无明显增加,与对照组相似,约为10%.动物体内实验表明,CD45-U266细胞群在SCID-hIL-6小鼠体内存活能力是CD45+U266细胞群的5倍.经DiOC6染色流式细胞术检测马法兰处理的U266细胞线粒体膜电位,CD45+U266细胞线粒体膜电位下降60%,而CD45-U266细胞线粒体膜电位仅下降30%,两对照组线粒体膜电位下降均为10%左右.经紫外线照射后,CD45+U266细胞更易从线粒体释放细胞色素C,导致更多的caspase-9被激活.结论 CD45分子的表达参与线粒体介导的细胞凋亡过程,使骨髓瘤细胞对多种凋亡刺激因子的敏感性增加.
목적 탐토CD45분자적표체여골수류세포조망적관계.방법 용마법란유도골수류세포계U266적조망;무혈청배양법유도전염CD45기인혹공질립적U266세포조망;무포도당배양법유도AMO1세포조망;동물실험비교U266세포CD45음성화양성량군세포재체내적존활능력;류식세포술검측조망세포솔화선립체막전위;Western blot법검측선립체내세포색소C적석방화caspase-9적격활.결과 경마법란처리후,45%적CD45+U266세포발생조망,이CD45-U266세포지유30%발생조망;재무포도당배양조건하,여저표체CD45적AMO1세포상비,고표체CD45적AMO1세포경역발생조망.무혈청배양5 d,전염CD45RB기인적U266세포60%발생조망,이전염공질립적U266세포적조망수무명현증가,여대조조상사,약위10%.동물체내실험표명,CD45-U266세포군재SCID-hIL-6소서체내존활능력시CD45+U266세포군적5배.경DiOC6염색류식세포술검측마법란처리적U266세포선립체막전위,CD45+U266세포선립체막전위하강60%,이CD45-U266세포선립체막전위부하강30%,량대조조선립체막전위하강균위10%좌우.경자외선조사후,CD45+U266세포경역종선립체석방세포색소C,도치경다적caspase-9피격활.결론 CD45분자적표체삼여선립체개도적세포조망과정,사골수류세포대다충조망자격인자적민감성증가.
Objective To investigate the effects of CD45 expression on induction of apoptosis in multiple myeloma cells. Methods Melphalan was used to induce myeloma cell line U266 apoptosis. Serum-free culture was used to induce CD45RB gene or empty plasmid transfected U266 apoptosis. The glucose-free culture was used to induce high CD45 (CD45h1) or low CD45 (CD45low) expression AMO1 apoptosis. Intraperitoneal inoculation was used to compare the survival of CD45 - or CD45 + U266 cells in mice. The number of apoptotic cells and mitochondrial membrane potential(MMP) was detected by flow cytometry. Western blotting was used to detect the cytochrome C release from mitochondrial and Caspase-9 activation. Results Melphalan treatment induced 45% of CD45 + and 30% of CD45 - U266 cells apoptosis. Compared with the CD45low AMO1 cells, CD45hl cells were more susceptible to apoptosis. In serum-free culture for five days,60% of CD45RB transferred U266 cells underwent apoptosis, while in the empty plasmid transfected ones,apoptotic cell number was not significantly increased. The survival time of CD45 - U266 cells in the SCID-hIL-6 mice was 5 times that of CD45 + cells. After melphalan treatment, 60% of the CD45 + U266 cells lost MMP, while only 36% of CD45 - U266 cells, and 10% of control cells did so. After UV irradiation, CD45 +U266 cells mitochondria released more cytochrome C, leading to more caspase-9 activation. Conclusion CD45 expression is involved in mitochondria-mediated apoptotic process and increases apoptotic sensitivity of myeloma cells under a variety of stimulation.