国际遗传学杂志
國際遺傳學雜誌
국제유전학잡지
INTERNATIONAL JOURNAL OF GENETICS
2011年
3期
130-134
,共5页
重组肿瘤抑素%细胞凋亡%线粒体膜电位%细胞色素C
重組腫瘤抑素%細胞凋亡%線粒體膜電位%細胞色素C
중조종류억소%세포조망%선립체막전위%세포색소C
Recombinant tumstatin%Apoptosis%Mitochondrial membrane potential%Cytochrome C
目的 检测重组肿瘤抑素42肽(T42)诱导肝癌HepG2细胞凋亡及与线粒体凋亡途径的关系,探讨T42诱导肿瘤细胞凋亡的可能机制.方法 吖啶橙/溴化乙锭(AO/EB)荧光染色观察细胞凋亡的形态学变化;流式细胞仪检测凋亡率;JC-1荧光染色检测线粒体膜电位的变化;Western印迹检测细胞色素C(Cyt-C)的分布.结果 18 μmol/LT42作用下,HepG2细胞出现明显凋亡形态学变化,凋亡率为22.4%,与对照组比较差异有统计学意义(t=7.75,P＜0.05);T42降低了HepG2细胞线粒体膜电势,明显减少了线粒体Cyt-C.结论 T42通过降低线粒体膜电势,促进Cyt-C由线粒体膜释放到胞浆中,激活 caspase-3途径诱导人肝癌细胞系HepG2细胞凋亡.
目的 檢測重組腫瘤抑素42肽(T42)誘導肝癌HepG2細胞凋亡及與線粒體凋亡途徑的關繫,探討T42誘導腫瘤細胞凋亡的可能機製.方法 吖啶橙/溴化乙錠(AO/EB)熒光染色觀察細胞凋亡的形態學變化;流式細胞儀檢測凋亡率;JC-1熒光染色檢測線粒體膜電位的變化;Western印跡檢測細胞色素C(Cyt-C)的分佈.結果 18 μmol/LT42作用下,HepG2細胞齣現明顯凋亡形態學變化,凋亡率為22.4%,與對照組比較差異有統計學意義(t=7.75,P＜0.05);T42降低瞭HepG2細胞線粒體膜電勢,明顯減少瞭線粒體Cyt-C.結論 T42通過降低線粒體膜電勢,促進Cyt-C由線粒體膜釋放到胞漿中,激活 caspase-3途徑誘導人肝癌細胞繫HepG2細胞凋亡.
목적 검측중조종류억소42태(T42)유도간암HepG2세포조망급여선립체조망도경적관계,탐토T42유도종류세포조망적가능궤제.방법 아정등/추화을정(AO/EB)형광염색관찰세포조망적형태학변화;류식세포의검측조망솔;JC-1형광염색검측선립체막전위적변화;Western인적검측세포색소C(Cyt-C)적분포.결과 18 μmol/LT42작용하,HepG2세포출현명현조망형태학변화,조망솔위22.4%,여대조조비교차이유통계학의의(t=7.75,P＜0.05);T42강저료HepG2세포선립체막전세,명현감소료선립체Cyt-C.결론 T42통과강저선립체막전세,촉진Cyt-C유선립체막석방도포장중,격활 caspase-3도경유도인간암세포계HepG2세포조망.
Objective The aim of the present article is to detect the apoptosis of hepatocarcinoma cells HepG2 induced by recombinant tumor endostatin 42 peptide (T42) ,with an emphasis on the signaling pathways involved. Methods Observed the morphological changes associated with the apoptosis of HepG2 cells by using AO/EB. Apoptosis rate were dentified by using flow cytometry. Mitochondrial membrane potential was evaluated by using JC-1 fluorescent staining. The distribution of cytochrome C(Cytc ) was estimated by using western blot. Results Compared with the control group, there was significant difference in apoptosis rate of cells HepG2 under 18μmol /L of T42. (22.4% vs 3.70% ,t =7.75, P＜0.05). Mitochondrial membrane potential was decreased by T42, and cytochrome c was reduced significantly compared with the control group. Conclusions The result demonstrated that the T42 enhanced the apoptosis of HepG2 cells and its potential mechanism was related to the decreased of mitochondrial membrane potential, an increase in Cytochrome C released into the cytosol, and reduced activation of Caspase-3 channels.
