国际泌尿系统杂志
國際泌尿繫統雜誌
국제비뇨계통잡지
International Journal of Urology and Nephrology
2015年
5期
674-677
,共4页
王伟%徐长庚%周向军%张杰
王偉%徐長庚%週嚮軍%張傑
왕위%서장경%주향군%장걸
膀胱肿瘤%茯苓酸
膀胱腫瘤%茯苓痠
방광종류%복령산
Urinary Bladder Neoplasms%PACHYMIC ACID
目的 探究茯苓酸(PA)对膀胱癌细胞BIU87的抑制作用及其机制.方法 体外培养BIU87细胞后加入不同浓度的茯苓酸,CCK-8法检测其对BIU87细胞的生存抑制作用.采用Hoechst染色法观察不同浓度下茯苓酸诱导细胞凋亡的作用.为进一步探究机制,将BIU87细胞采用20μg/mL、40μg/mL、80 μg,/mL的茯苓酸处理后,检测Caspase3的活性及采用westem blot检测凋亡特异性蛋白Caspase9、Cleaved-caspase9、Bcl-2、Bax、Bak的表达.结果 茯苓酸对BIU87细胞有明显的抑制作用,且呈现剂量依赖的趋势.20μg/mL、40μg/mL、80μg/mL的PA作用BIU87细胞后可产生明显的细胞凋亡作用,Hoechst染色可见明显的核碎裂和凋亡小体.计数阳性细胞数目分别为20μg/mL(6.7±1.5)、40μg/mL (13.3±3.5)、80μg/mL(20.7±1.5).BIU87细胞经茯苓酸处理后,Caspase3活化水平明显升高,以对照组为标准,20μg/mL组为(167.3±18.7)％、40μg/mL为(215.0±26.9)％、80μg/mL为(289.7±17.1)％.且Cleaved-caspase9、Bax、Bak蛋白表达水平逐渐升高,Caspase9、Bcl-2蛋白表达逐渐降低,均呈浓度依赖的趋势.结论 茯苓酸可通过诱导细胞凋亡对膀胱癌细胞BIU87产生抑制作用,且可能是通过内源性线粒体凋亡途径介导实现的.
目的 探究茯苓痠(PA)對膀胱癌細胞BIU87的抑製作用及其機製.方法 體外培養BIU87細胞後加入不同濃度的茯苓痠,CCK-8法檢測其對BIU87細胞的生存抑製作用.採用Hoechst染色法觀察不同濃度下茯苓痠誘導細胞凋亡的作用.為進一步探究機製,將BIU87細胞採用20μg/mL、40μg/mL、80 μg,/mL的茯苓痠處理後,檢測Caspase3的活性及採用westem blot檢測凋亡特異性蛋白Caspase9、Cleaved-caspase9、Bcl-2、Bax、Bak的錶達.結果 茯苓痠對BIU87細胞有明顯的抑製作用,且呈現劑量依賴的趨勢.20μg/mL、40μg/mL、80μg/mL的PA作用BIU87細胞後可產生明顯的細胞凋亡作用,Hoechst染色可見明顯的覈碎裂和凋亡小體.計數暘性細胞數目分彆為20μg/mL(6.7±1.5)、40μg/mL (13.3±3.5)、80μg/mL(20.7±1.5).BIU87細胞經茯苓痠處理後,Caspase3活化水平明顯升高,以對照組為標準,20μg/mL組為(167.3±18.7)％、40μg/mL為(215.0±26.9)％、80μg/mL為(289.7±17.1)％.且Cleaved-caspase9、Bax、Bak蛋白錶達水平逐漸升高,Caspase9、Bcl-2蛋白錶達逐漸降低,均呈濃度依賴的趨勢.結論 茯苓痠可通過誘導細胞凋亡對膀胱癌細胞BIU87產生抑製作用,且可能是通過內源性線粒體凋亡途徑介導實現的.
목적 탐구복령산(PA)대방광암세포BIU87적억제작용급기궤제.방법 체외배양BIU87세포후가입불동농도적복령산,CCK-8법검측기대BIU87세포적생존억제작용.채용Hoechst염색법관찰불동농도하복령산유도세포조망적작용.위진일보탐구궤제,장BIU87세포채용20μg/mL、40μg/mL、80 μg,/mL적복령산처리후,검측Caspase3적활성급채용westem blot검측조망특이성단백Caspase9、Cleaved-caspase9、Bcl-2、Bax、Bak적표체.결과 복령산대BIU87세포유명현적억제작용,차정현제량의뢰적추세.20μg/mL、40μg/mL、80μg/mL적PA작용BIU87세포후가산생명현적세포조망작용,Hoechst염색가견명현적핵쇄렬화조망소체.계수양성세포수목분별위20μg/mL(6.7±1.5)、40μg/mL (13.3±3.5)、80μg/mL(20.7±1.5).BIU87세포경복령산처리후,Caspase3활화수평명현승고,이대조조위표준,20μg/mL조위(167.3±18.7)％、40μg/mL위(215.0±26.9)％、80μg/mL위(289.7±17.1)％.차Cleaved-caspase9、Bax、Bak단백표체수평축점승고,Caspase9、Bcl-2단백표체축점강저,균정농도의뢰적추세.결론 복령산가통과유도세포조망대방광암세포BIU87산생억제작용,차가능시통과내원성선립체조망도경개도실현적.
Objectives The article aims to detect the inhibition effect of bladder cancer cells treated with pachymic acid and its possible mechanisms.Methods CCK-8 assay was applied to measure the cell viability after cells incubated with pachymic acid.After that,morphology of apoptosis was observed by Hoechst staining under different concentrations of PA.To further identify a possible mechanism,intrinsic apoptosis related protein levels of Cleaved-caspase9,Caspase9,Bax,Bak and Bcl-2 were detected by Western-Blot.Results PA significant inhibited the viability of BIU87 cells in a dose-dependent manners;Meanwhile,The BIU87 cells present obvious apoptosis and apoptosis rate reaches to 39.1％ (P ＜ 0.05).A notable apoptotic effect was observed after when it was treated with 20μg/mL PA,40ug/ml and 80ug/mL.In addition,Hoechst staining showed nuclear fragmentation and apoptotic bodies.The numbers of Hoechst staining positive cells in each group were:20μg/mL(6.7 ± 1.5),40μg/ mL(13.3 ± 3.5),80μg/mL(20.7 ± 1.5).Furthermore,activation of Caspase3 levels were significant increased after the treatment.Take the control group as the standard,the Caspase3 activation levels in three groups were:(167.3 ± 18.7)％,40μg/mL(215.0 ± 26.9)％,80μg/mL(289.7 ± 17.1)％.The protein expressions of Cleaved-caspase9,Bax and Bak were up-regulated and had a dose-dependent tendency while Bcl-2 and Caspase9 were down-regulated.Conclusions PA has an inhibited effect on bladder cancer cells BIU87 by inducing apoptosis,which probably through an intrinsic apoptotic pathway.
