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[柚皮素体外诱导K562细胞凋亡与半胱天冬酶-3和半胱天冬酶-8酶活性变化及FAS/FASL蛋白表达的关系]

[Relation of apoptosis of K562 cells induced by naringenin in vitro to enzyme activity changes of caspase-3 and caspase-8 and expression of FAS/FASL proteins].

作者信息

Zuo Xue-Lan, Zhou Ying, Li Rui-Fang, Feng Ying-Qian, He Li, Liu Ming-Hui

机构信息

Department of Hematology, Zhongnan Hospital, Wuhan University, Wuhan 430071, Hubei Province, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2008 Apr;16(2):286-9.

Abstract

The objective of this study was to investigate the apoptosis-inducing effect and underlying mechanism of naringenin (NGEN) on K562 cells in vitro. The inhibition of NGEN on K562 cells was evaluated by means of MTT assay so as to observe the cytotoxicity of NGEN; The morphological changes of the cells treated by NGEN were observed by transmission electron microscope; cell apoptosis rate influenced by NGEN was assessed by flow cytometry; the enzyme activity changes of caspase-3 and caspase-8 in the process of NGEN-induced K562 apoptosis were detected by Caspase Colorimetric Assay Kit; immunohistochemistry technique was used to detect FAS, FASL protein expression in K562 cells. The results showed that the growth of cells was inhibited by NGEN in dose-and time-dependent manners (p<0.05); NGEN-induced K562 cells apoptosis and sub-G1 peak was observed; some typically early and final phase changes of cell apoptosis were revealed under transmission electron microscope; the enzyme activity of caspase-3 and caspase-8 and the expression of FAS remarkably increased, meanwhile the expression of FASL was down-regulated (p<0.05). It is concluded that NGEN exerts anti-cancer effect by inducing K562 cell apoptosis, and the regulation of the expression of FAS and FASL. The caspase-3 and caspase-8 co-pathway brings about one of the mechanisms.

摘要

本研究的目的是探讨柚皮素(NGEN)体外对K562细胞的诱导凋亡作用及其潜在机制。采用MTT法评价NGEN对K562细胞的抑制作用,以观察NGEN的细胞毒性;通过透射电子显微镜观察NGEN处理后细胞的形态变化;采用流式细胞术评估NGEN对细胞凋亡率的影响;用Caspase比色法检测NGEN诱导K562细胞凋亡过程中caspase-3和caspase-8的酶活性变化;采用免疫组织化学技术检测K562细胞中FAS、FASL蛋白表达。结果显示,NGEN以剂量和时间依赖性方式抑制细胞生长(p<0.05);观察到NGEN诱导K562细胞凋亡及亚G1峰;透射电子显微镜下显示出一些典型的细胞凋亡早期和终末期变化;caspase-3和caspase-8的酶活性及FAS表达显著增加,同时FASL表达下调(p<0.05)。结论是,NGEN通过诱导K562细胞凋亡及调节FAS和FASL表达发挥抗癌作用。caspase-3和caspase-8共同通路是其中一种机制。

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