摘要
目的探讨中华猕猴桃根乙酸乙酯提取部分(EE-ACP)对SW480的增殖抑制及诱导凋亡的作用及其可能的机制。方法运用MTT比色法、Hochest33258染色法、流式细胞术和Western Blot法检测EE-ACP对SW480细胞的作用。结果不同浓度的EE-ACP作用于SW480细胞在24、48、72 h后均能明显的抑制细胞的增殖(P<0.05),且存在时间-浓度依赖性,IC50分别为224.344、195.376和160.459μg/ml。Hochest33258荧光染色可见不同浓度EE-CAR干预SW480细胞48 h后,凋亡小体逐渐增多。FITC-Annexin V/PI双染法提示EE-ACP诱导细胞凋亡,并随剂量的增加作用增强。PI染色结果提示,EE-ACP将SW480细胞周期阻滞于G_1期。Western Blot结果显示,随着加药浓度的增加,周期相关蛋白P53和P21表达逐渐增大,周期相关蛋白Cyclin D1表达减少。结论中华猕猴桃根乙酸乙酯提取部分可以抑制SW480细胞的增值,其发生可能与其促进P53、P21基因,抑制Cyclin D1基因表达有关。
Objective To investigate the inhibition efficacy of Actinidiachinensis planch extracted by ethyl acetate（ EE-ACP） on proliferation of human cancer cell line SW480 cells and induction on apoptosis,and to explore its mechanism. Methods Using the MTT assay,Hochest33258 stain,flow cytometric（ FCM） analysis and Western Blot methods,the EE-ACP＇s affects on SW480 cells were detected in this study. Results The result of MTT showed that,after EE-ACP treatment,the proliferation of SW480 cells were obviously inhibited in a dose-and time-dependent manner. The IC50 was 224. 344,195. 376 and 60. 459 μg / ml. The result of hochest33258 showed that,after treated by EE-ACP,the apoptosis morphology of SW480 cells was observed. The apoptotic cells were detected after treated with 80,160 and 320μg / ml of the EE-ACP for 48 hours and the effect was enhanced as the dosage increased. The percentages of the G0 / G1 phase cells were increased and S phase cells were decreased after treated by EE-ACP for 48 hours. Apoptptic protein P53 and cycly protein P21 were up-regulated,while cyclin D1 protein down-regulated. Conclusion EE-ACP inhibits the proliferation of SW480 cells,probably through the up-regulation of P53 and P21,and down-regulation of cyclin D1.
出处
《临床军医杂志》
CAS
2016年第1期55-59,共5页
Clinical Journal of Medical Officers
基金
广西高等学校重点资助基金(210202ZD064)
