You Position: Home > Paper
Apoptosis of hepatocelluar carcinoma BEL-7402 cells in vitro induced by rapamycin and its action mechanism
( views:183, downloads:8 )
- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF ORGAN TRANSPLANTATION
- Issue:
- 10
- DOI:
- 10.3760/cma.j.issn.0254-1785.2006.10.002
- Key Word:
- 西罗莫司;肝肿瘤;细胞凋亡;线粒体
Abstract： 目的 探讨西罗莫司(SRL)在体外诱导人肝癌细胞株BEL-7402的凋亡作用及其机制.方法 以不同浓度的SRL(5、10、20、30、40和50nmol/L)作用于体外培养的人肝癌细胞株BEL-7402,应用流式细胞仪检测培养24、48和72 h时的细胞凋亡情况;Hoechst 33258荧光染色法观察细胞凋亡时的形态学变化;Western Blot法观察BEL-7402细胞中Caspase-3、Bcl-2、Bcl-xl和Bax基因的表达变化;采用Caspase-3试剂盒检测Caspase-3酶的活性;JC-1染色法检测细胞线粒体膜电位(△ψm).结果 SRL可诱导BEL-7402细胞凋亡,并与药物浓度和作用时间呈正相关.SRL作用BEL-7402细胞后48 h,在Hoechst 33258荧光染色图片上可见核浓缩及核碎裂等典型的细胞凋亡特征,凋亡过程中线粒体膜电位下降及Caspase-3酶原蛋白激活,伴有Bcl-2、Bcl-x蛋白表达的降低和Bax蛋白上调.结论 SRL能使抗凋亡蛋白Bcl-2、Bcl-xl表达降低,促凋亡蛋白Bax表达上调,线粒体膜电位下降,激活Caspase-3酶原蛋白,从而诱导细胞凋亡.
- [1]D D, Newmeyer,D M, Farschon,J C, Reed.Cell-free apoptosis in Xenopus egg extracts: inhibition by Bcl-2 and requirement for an organelle fraction enriched in mitochondria.[J].1994,79(2).
- [2]S, Krajewski,M, Krajewska,A, Shabaik,etc.Immunohistochemical determination of in vivo distribution of Bax, a dominant inhibitor of Bcl-2.[J].1994,145.
- [3]Granger DK,Canafax DM,Chen SC,etc.Prolongation of renal allograft survival in a large animal model by oral rapamycin monotherapy.[J].1995,59(2).
- [4]Kreis Henri,Cisterne Jean-Marc,Land Walter,etc.Sirolimus in Association with Mycophenolate Mofetil Induction for the Prevention of Acute Graft Rejection in Renal Allograft Recipients12[J].2000.
- [5]M O, Hengartner.The biochemistry of apoptosis.[J].2000,407(6805).
- [6]Dirksen RobertT,Sheu SheyShing,Beutner Gisela,etc.Mitochondrial permeability transition and calcium dynamics in striatal neurons upon intense NMDA receptor activation.[J].2002,80(3).
- [7]Rini BI,Wright TM,Rathmell WK.Molecularly targeted therapy in renal cell carcinoma.[J].2005,5(6).
- [8]S N, Farrow,J H, White,I, Martinou,etc.Cloning of a bcl-2 homologue by interaction with adenovirus E1B 19K.[J].1995,374(6524).