中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
2期
252-254
,共3页
冀保卫%陈谦学%田道锋%吴立权%刘宝辉%郭振涛%纪振刚%朱晓楠
冀保衛%陳謙學%田道鋒%吳立權%劉寶輝%郭振濤%紀振剛%硃曉楠
기보위%진겸학%전도봉%오립권%류보휘%곽진도%기진강%주효남
胶质瘤%STAT3%脱噬作用
膠質瘤%STAT3%脫噬作用
효질류%STAT3%탈서작용
Glioma%STAT3%Apoptosis
目的 探讨组蛋白去乙酰化酶抑制剂MS-275对脑胶质瘤细胞株U251细胞增殖的影响及促凋亡机制.方法 以不同药物浓度梯度和U251细胞分别共培养24、48、72 h后,应用CCK-8法检测肿瘤细胞增殖,瑞氏-姬姆萨染色法观察细胞形态变化,PI单染法检测细胞周期,Annexin V-FITC/PI法经流式检测细胞凋亡率,免疫印迹检测STAT3蛋白和PARP蛋白表达.结果 MS-275能显著抑制U251细胞的增殖,药物浓度40 nmol/ml,培养72 h,抑制率为(79.0±1.7)%;经药物作用后,细胞呈现凋亡形态,胞质和胞核浓缩或碎裂;G0/G1期细胞由(66.320±0.456)%降至(38.288±1.242)%(P＜0.01),G2/M期细胞由(19.940±0.580)%升高至(35.650±0.507)%,而后降至(22.343±1.224)%(P＜0.01),亚G0凋亡峰由(0.230±0.035)%增高至(18.393±1.449)%(P＜0.01);总凋亡率由(2.133±0.416)%增高至(29.000±2.307)%(P＜0.01);免疫印迹检测提示磷酸化STAT3表达水平降低,PARP被剪切.结论 MS-275对胶质瘤细胞的增殖抑制作用具有浓度和时间依赖性,它通过阻断STAT3磷酸化,诱导激活Caspase-3凋亡途径.
目的 探討組蛋白去乙酰化酶抑製劑MS-275對腦膠質瘤細胞株U251細胞增殖的影響及促凋亡機製.方法 以不同藥物濃度梯度和U251細胞分彆共培養24、48、72 h後,應用CCK-8法檢測腫瘤細胞增殖,瑞氏-姬姆薩染色法觀察細胞形態變化,PI單染法檢測細胞週期,Annexin V-FITC/PI法經流式檢測細胞凋亡率,免疫印跡檢測STAT3蛋白和PARP蛋白錶達.結果 MS-275能顯著抑製U251細胞的增殖,藥物濃度40 nmol/ml,培養72 h,抑製率為(79.0±1.7)%;經藥物作用後,細胞呈現凋亡形態,胞質和胞覈濃縮或碎裂;G0/G1期細胞由(66.320±0.456)%降至(38.288±1.242)%(P＜0.01),G2/M期細胞由(19.940±0.580)%升高至(35.650±0.507)%,而後降至(22.343±1.224)%(P＜0.01),亞G0凋亡峰由(0.230±0.035)%增高至(18.393±1.449)%(P＜0.01);總凋亡率由(2.133±0.416)%增高至(29.000±2.307)%(P＜0.01);免疫印跡檢測提示燐痠化STAT3錶達水平降低,PARP被剪切.結論 MS-275對膠質瘤細胞的增殖抑製作用具有濃度和時間依賴性,它通過阻斷STAT3燐痠化,誘導激活Caspase-3凋亡途徑.
목적 탐토조단백거을선화매억제제MS-275대뇌효질류세포주U251세포증식적영향급촉조망궤제.방법 이불동약물농도제도화U251세포분별공배양24、48、72 h후,응용CCK-8법검측종류세포증식,서씨-희모살염색법관찰세포형태변화,PI단염법검측세포주기,Annexin V-FITC/PI법경류식검측세포조망솔,면역인적검측STAT3단백화PARP단백표체.결과 MS-275능현저억제U251세포적증식,약물농도40 nmol/ml,배양72 h,억제솔위(79.0±1.7)%;경약물작용후,세포정현조망형태,포질화포핵농축혹쇄렬;G0/G1기세포유(66.320±0.456)%강지(38.288±1.242)%(P＜0.01),G2/M기세포유(19.940±0.580)%승고지(35.650±0.507)%,이후강지(22.343±1.224)%(P＜0.01),아G0조망봉유(0.230±0.035)%증고지(18.393±1.449)%(P＜0.01);총조망솔유(2.133±0.416)%증고지(29.000±2.307)%(P＜0.01);면역인적검측제시린산화STAT3표체수평강저,PARP피전절.결론 MS-275대효질류세포적증식억제작용구유농도화시간의뢰성,타통과조단STAT3린산화,유도격활Caspase-3조망도경.
Objective To investigate the effect of MS-275 on the proliferation of brain glioma cells. Methods U251 cells were respectively cultured for 24, 48, 72 h, with different concentrations of MS-275. CCK-8 assay was used to assess the proliferation of U251 cells. The morphological changes of U251 cells were observed by Wright-Giemsa staining. The cell cycle was analyzed by PI dyeing. The apoptosis rate was assayed by flow cytometry using annexin V-FITC/PI. The protein expression of STAT3 and PARP was detected by Western blotting. Results MS-275 could significantly inhibited proliferation of U251 cells. After treatment with MS-275, apoptosis of U251 cells was seen; The ratio of G0/G1 was decreased from ( 66.320 ± 0.456 ) % to ( 38. 288 ± 1. 242 ) % ( P ＜ 0. 01 ), the percentage of G2/M cells was increased from ( 19. 940 ± 0. 580 ) % to ( 35.650 ± 0. 507 ) %, then decreased to ( 22. 343 ± 1. 224 ) %(P＜0.01), and ratio of sub-G0 was increased from (0.230 ±0.035)% to (18.393 ±1.449)% (P＜0. 01 ); Total apoptosis rate was increased from ( 2. 133 ± 0.416 ) % to ( 29. 000 ± 2. 307 ) % ( P ＜ 0. 01 );The expression level of phosphorylated STAT3 was significantly downregulated, and the cleavage of PARP was detected by Western blotting. Conclusion MS-275 inhibites proliferation of brain glioma cells in a time- and dose-dependent manner, which is achieved by inducing apoptosis.
