中华放射医学与防护杂志
中華放射醫學與防護雜誌
중화방사의학여방호잡지
Chinese Journal of Radiological Medicine and Protection
2014年
7期
497-501
,共5页
姜新%刘野%常鹏宇%曲雅勤%黄巍%王立哲%蒋鑫萍%Rui Ji%辛颖
薑新%劉野%常鵬宇%麯雅勤%黃巍%王立哲%蔣鑫萍%Rui Ji%辛穎
강신%류야%상붕우%곡아근%황외%왕립철%장흠평%Rui Ji%신영
渥曼青霉素%PI3K信号通路%恶性胶质瘤%辐射敏感性%线粒体凋亡途径
渥曼青黴素%PI3K信號通路%噁性膠質瘤%輻射敏感性%線粒體凋亡途徑
악만청매소%PI3K신호통로%악성효질류%복사민감성%선립체조망도경
Wortmannin%PI3K signaling pathway%Malignant glioma%Radiation sensitivity%Mitochondria apoptosis pathway
目的利用渥曼青霉素(wortmannin,WM)抑制人恶性胶质瘤细胞U251磷脂酰肌醇-3-激酶(PI3K)通路的活性,探讨对U251细胞辐射敏感性的影响及可能的作用机制.方法 采用10μmol/L WM预处理U251 2 h,并接受10 Gy X射线照射,检测PI3K/Akt信号通路的活性、集落形成率和凋亡的变化;通过Western blot方法检测凋亡相关蛋白活化型Caspase-3、Bax、Bcl-2及XIAP表达的变化.结果 10 μmol/L WM预处理2h,明显抑制了U251细胞phospho-Akt的表达(t=0.000 1,P＜0.01).WM预处理联合X射线照射后,U251细胞的凋亡率由对照组和单纯照射组的(2.14±1.32)％和(11.5±2.9)％增加到(22.6±3.8)％,差异具有统计学意义(t=0.009 3、0.002 7,P＜0.01);集落形成率由对照组和单纯照射组的(88.54±4.76)％和(56.31±4.05)％降低到(12.25±9.59)％(t=0.000 03、0.000 2,P＜0.01);同时伴随着凋亡相关蛋白活化型Caspase-3表达显著增加,Bax/Bcl-2的比值明显增高以及XIAP的显著降低.结论 WM通过抑制PI3 K/Akt信号通路活性,增加凋亡蛋白Caspase-3的活化和Bax/Bcl-2比值,下调凋亡抑制蛋白XIAP来增强恶性胶质瘤细胞的辐射敏感性.
目的利用渥曼青黴素(wortmannin,WM)抑製人噁性膠質瘤細胞U251燐脂酰肌醇-3-激酶(PI3K)通路的活性,探討對U251細胞輻射敏感性的影響及可能的作用機製.方法 採用10μmol/L WM預處理U251 2 h,併接受10 Gy X射線照射,檢測PI3K/Akt信號通路的活性、集落形成率和凋亡的變化;通過Western blot方法檢測凋亡相關蛋白活化型Caspase-3、Bax、Bcl-2及XIAP錶達的變化.結果 10 μmol/L WM預處理2h,明顯抑製瞭U251細胞phospho-Akt的錶達(t=0.000 1,P＜0.01).WM預處理聯閤X射線照射後,U251細胞的凋亡率由對照組和單純照射組的(2.14±1.32)％和(11.5±2.9)％增加到(22.6±3.8)％,差異具有統計學意義(t=0.009 3、0.002 7,P＜0.01);集落形成率由對照組和單純照射組的(88.54±4.76)％和(56.31±4.05)％降低到(12.25±9.59)％(t=0.000 03、0.000 2,P＜0.01);同時伴隨著凋亡相關蛋白活化型Caspase-3錶達顯著增加,Bax/Bcl-2的比值明顯增高以及XIAP的顯著降低.結論 WM通過抑製PI3 K/Akt信號通路活性,增加凋亡蛋白Caspase-3的活化和Bax/Bcl-2比值,下調凋亡抑製蛋白XIAP來增彊噁性膠質瘤細胞的輻射敏感性.
목적이용악만청매소(wortmannin,WM)억제인악성효질류세포U251린지선기순-3-격매(PI3K)통로적활성,탐토대U251세포복사민감성적영향급가능적작용궤제.방법 채용10μmol/L WM예처리U251 2 h,병접수10 Gy X사선조사,검측PI3K/Akt신호통로적활성、집락형성솔화조망적변화;통과Western blot방법검측조망상관단백활화형Caspase-3、Bax、Bcl-2급XIAP표체적변화.결과 10 μmol/L WM예처리2h,명현억제료U251세포phospho-Akt적표체(t=0.000 1,P＜0.01).WM예처리연합X사선조사후,U251세포적조망솔유대조조화단순조사조적(2.14±1.32)％화(11.5±2.9)％증가도(22.6±3.8)％,차이구유통계학의의(t=0.009 3、0.002 7,P＜0.01);집락형성솔유대조조화단순조사조적(88.54±4.76)％화(56.31±4.05)％강저도(12.25±9.59)％(t=0.000 03、0.000 2,P＜0.01);동시반수착조망상관단백활화형Caspase-3표체현저증가,Bax/Bcl-2적비치명현증고이급XIAP적현저강저.결론 WM통과억제PI3 K/Akt신호통로활성,증가조망단백Caspase-3적활화화Bax/Bcl-2비치,하조조망억제단백XIAP래증강악성효질류세포적복사민감성.
Objective To explore the effect of wortmannin (WM),an inhibitor of PI3K signalingpathway,on the radiation sensitivity of human malignant glioma cells (U251) and the underlyingmechanism.Methods U251 cells were pretreated with 10 μmol/L WM for 2 h and irradiated with 10 GyX-rays,then the activity of PI3K/Akt signaling pathway,the colony-forming ability and apoptosis weredetected to evaluate the effect of WM on radiosensitivity.The protein expressions of cleaved Caspase-3,Bax,Bcl-2 and XIAP were detected with Western blot assay.Results The expression of phospho-Akt wassignificantly inhibited by the pretreatment of cells with 10 μ mol/L WM (t =0.000 1,P ＜0.01).The ratioof apoptotic cells in the WM + IR group was significantly increased from (2.14 ± 1.32)％ of control groupand (11.5 ±2.9)％ of IR group to (22.6 ±3.8)％ (t =0.009 3,0.002 7,P＜0.01).The colony-forming rate of WM + IR group was decreased from (88.54 ± 4.76) ％ of control group and (56.31 ±4.05)％ of IR group to (12.25 ±9.59)％ significantly (t=0.000 03,0.000 2,P＜0.01) accompaniedwith the increases of Caspase-3 and Bax/Bcl-2 but decrease of XIAP.Conclusions WM significantlyincreased the radiation sensitivity of human malignant glioma cells by inhibiting the activity of PI3Ksignaling pathway and the expressions of Bcl-2 and XIAP and activating Caspase-3 and Bax.
