中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2012年
1期
43-46
,共4页
邵化敏%唐宇宏%沈群%朱红青%季鸥%章亚成%季建敏%姜鹏君%司叶俊%李兆荣
邵化敏%唐宇宏%瀋群%硃紅青%季鷗%章亞成%季建敏%薑鵬君%司葉俊%李兆榮
소화민%당우굉%침군%주홍청%계구%장아성%계건민%강붕군%사협준%리조영
葛根总黄酮%Kasumi-1细胞%融合蛋白质类,AML1-ETO%细胞凋亡
葛根總黃酮%Kasumi-1細胞%融閤蛋白質類,AML1-ETO%細胞凋亡
갈근총황동%Kasumi-1세포%융합단백질류,AML1-ETO%세포조망
Puerariae radix flavones(PRF)%Kasumi-1 cells%Fusion proteins,AML1-ETO%Apoptosis
目的 探讨葛根总黄酮(puerariae radix flavones,PRF)对人急性髓系白血病细胞系Kasumi-1细胞凋亡的影响,并对其可能的分子机制进行初步研究.方法 以不同浓度PRF作用Kasumi-1细胞48 h,瑞氏染色及Hoechst 33258荧光染色观察细胞凋亡变化,FITC-Annexin V/PI双染法检测细胞凋亡率,实时定量PCR技术检测AMLl -ETO融合基因表达,Western blot法检测Bcl-2、Bim及Caspase相关酶的变化.结果 PRF能有效诱导Kasumi-1细胞凋亡.以50、200及500 μg/ml的PRF分别处理细胞,其早期凋亡率依次为(14.1±0.8)％、(17.7±1.3)％及(32.4±1.4)％,较空白对照组[(7.8±0.7)％]明显增加(P＜0.05),作用呈浓度依赖性；抗凋亡蛋白Bcl-2相对表达量依次为0.85±0.05、0.62±0.07及0.43±0.05,呈下调趋势(P＜0.01)；而促凋亡蛋白Bim相对表达量分别为0.21±0.06、0.39±0.04及0.75 +0.05,Caspase-3蛋白相对表达量分别为0.92±0.04、1.21±0.07及1.33±0.04,Caspase-9蛋白相对表达量分别为0.35±0.05、0.53±0.03及0.69±0.07,均呈上调趋势(P＜0.01).Caspase-8蛋白表达量及AML1 -ETO融合基因表达量则无明显改变.结论 一定浓度PRF诱导Kasumi-1细胞凋亡可能与下调细胞Bcl-2、上调Bim及活化Caspase相关酶有关,与AML1 -ETO融合基因无明显相关性.
目的 探討葛根總黃酮(puerariae radix flavones,PRF)對人急性髓繫白血病細胞繫Kasumi-1細胞凋亡的影響,併對其可能的分子機製進行初步研究.方法 以不同濃度PRF作用Kasumi-1細胞48 h,瑞氏染色及Hoechst 33258熒光染色觀察細胞凋亡變化,FITC-Annexin V/PI雙染法檢測細胞凋亡率,實時定量PCR技術檢測AMLl -ETO融閤基因錶達,Western blot法檢測Bcl-2、Bim及Caspase相關酶的變化.結果 PRF能有效誘導Kasumi-1細胞凋亡.以50、200及500 μg/ml的PRF分彆處理細胞,其早期凋亡率依次為(14.1±0.8)％、(17.7±1.3)％及(32.4±1.4)％,較空白對照組[(7.8±0.7)％]明顯增加(P＜0.05),作用呈濃度依賴性；抗凋亡蛋白Bcl-2相對錶達量依次為0.85±0.05、0.62±0.07及0.43±0.05,呈下調趨勢(P＜0.01)；而促凋亡蛋白Bim相對錶達量分彆為0.21±0.06、0.39±0.04及0.75 +0.05,Caspase-3蛋白相對錶達量分彆為0.92±0.04、1.21±0.07及1.33±0.04,Caspase-9蛋白相對錶達量分彆為0.35±0.05、0.53±0.03及0.69±0.07,均呈上調趨勢(P＜0.01).Caspase-8蛋白錶達量及AML1 -ETO融閤基因錶達量則無明顯改變.結論 一定濃度PRF誘導Kasumi-1細胞凋亡可能與下調細胞Bcl-2、上調Bim及活化Caspase相關酶有關,與AML1 -ETO融閤基因無明顯相關性.
목적 탐토갈근총황동(puerariae radix flavones,PRF)대인급성수계백혈병세포계Kasumi-1세포조망적영향,병대기가능적분자궤제진행초보연구.방법 이불동농도PRF작용Kasumi-1세포48 h,서씨염색급Hoechst 33258형광염색관찰세포조망변화,FITC-Annexin V/PI쌍염법검측세포조망솔,실시정량PCR기술검측AMLl -ETO융합기인표체,Western blot법검측Bcl-2、Bim급Caspase상관매적변화.결과 PRF능유효유도Kasumi-1세포조망.이50、200급500 μg/ml적PRF분별처리세포,기조기조망솔의차위(14.1±0.8)％、(17.7±1.3)％급(32.4±1.4)％,교공백대조조[(7.8±0.7)％]명현증가(P＜0.05),작용정농도의뢰성；항조망단백Bcl-2상대표체량의차위0.85±0.05、0.62±0.07급0.43±0.05,정하조추세(P＜0.01)；이촉조망단백Bim상대표체량분별위0.21±0.06、0.39±0.04급0.75 +0.05,Caspase-3단백상대표체량분별위0.92±0.04、1.21±0.07급1.33±0.04,Caspase-9단백상대표체량분별위0.35±0.05、0.53±0.03급0.69±0.07,균정상조추세(P＜0.01).Caspase-8단백표체량급AML1 -ETO융합기인표체량칙무명현개변.결론 일정농도PRF유도Kasumi-1세포조망가능여하조세포Bcl-2、상조Bim급활화Caspase상관매유관,여AML1 -ETO융합기인무명현상관성.
Objective To explore the effects and the molecular mechanism of puerariae radix flavones(PRF) on acute myeloid leukemia cell line Kasumi-1 cells in vitro.Methods Kasumi-1 cells treated by PRF for 48 hours were observed with Wright' s and Hoechst 33258 dying.The apoptotic cells were analyzed by flow cytometry with AnnexinV/PI staining.The expression levels of bcl-2,Bim and Caspase-3/-8/-9 protein were assayed by Western blot and the AML1-ETO fusion gene was detected by real-time polymerase chain reaction.Results PRF could induce Kasumi-1 cells to apoptosis effectively.The proportion of apoptotic cells in 50,200 and 500 μg/ml PRF treatment groups were ( 14.1 +0.8)％,( 17.7 + 1.3)％ and (32.4 ± 1.4) ％,respectively,and significantly higher than that of control [ ( 7.8 + 0.7 ) ％ ].The relative expression levels of the anti-apoptotic Bcl-2 protein were 0.85 ± 0.05,0.62 ± 0.07 and 0.43 ± 0.05 ; the apoptotic Bim protein were 0.21 ± 0.06,0.39 + 0.04 and 0.75 ± 0.05 ; the caspase-3 and caspase-9 were 0.92 ± 0.04,1.21 ± 0.07,1.33 ± 0.04 and 0.35 ± 0.05,0.53 ± 0.03,0.69 ± 0.07,respectively.Compared to the blank control group,all these changes were significant ( P ＜ 0.01 ).Nevertheless,nearly no changes could be observed on the expression level of AML1-ETO fusion gene and caspase-8 protein.Conclusion Apoptosis of Kasumi-1 cells induced by PRF might correlate to the down-regulation of Bcl-2 protein expression and the activation of caspase-3 and caspase-8 protein in the cells.It seemed that all these effects had no relationship with the AMLI-ETO fusion gene.
