中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2010年
12期
1785-1787
,共3页
王喻%李小娟%叶春伟%朱宝益%阮星星%陈泽斌%冯淑君%陶奕然%高新%温星桥
王喻%李小娟%葉春偉%硃寶益%阮星星%陳澤斌%馮淑君%陶奕然%高新%溫星橋
왕유%리소연%협춘위%주보익%원성성%진택빈%풍숙군%도혁연%고신%온성교
前列腺癌%氧化应激%生育酚结合蛋白%脱噬作用
前列腺癌%氧化應激%生育酚結閤蛋白%脫噬作用
전렬선암%양화응격%생육분결합단백%탈서작용
Prostate carcinoma%Oxidative stress%Tocopherol associated protein%Apoptosis
目的 观察生育酚结合蛋白(TAP)对氧化应激所致前列腺癌细胞凋亡的调控作用.方法 脂质体法分别转染pEGFP-N3-TAP质粒及空载体质粒进入前列腺癌细胞PC-3.以过氧化氢(H2O2)刺激PC-3细胞构建氧化应激模型,逆转录-聚合酶链反应(RT-PCR)、Western blot检测TAP的表达,流式细胞技术检测各组细胞的凋亡.结果转染TAP后的PC-3细胞中TAP表达量明显升高.RT-PCR结果显示,H2O2单独或联合促氧化剂丁胱亚磺酰亚胺(BSO)刺激细胞后TAP的mRNA表达量分别为(2.457±0.113)、(3.643±0.162)较对照组(0.721±0.048)显著上升,差异有统计学意义(P<0.01);在无H2O2刺激的对照组PC-3、PC-3-vector、PC-3-TAP细胞的凋亡率分别为2.42%、3.69%、15.37%;在H2O2刺激下,各组凋亡率分别为7.56%、16.71%、43.93%;联合应用BSO与H2O2,各组凋亡分别为13.73%、26.36%、49.19%.结论 TAP基因可参与前列腺癌细胞对氧化应激的应答反应,并促进氧化应激所致的前列腺癌细胞凋亡.
目的 觀察生育酚結閤蛋白(TAP)對氧化應激所緻前列腺癌細胞凋亡的調控作用.方法 脂質體法分彆轉染pEGFP-N3-TAP質粒及空載體質粒進入前列腺癌細胞PC-3.以過氧化氫(H2O2)刺激PC-3細胞構建氧化應激模型,逆轉錄-聚閤酶鏈反應(RT-PCR)、Western blot檢測TAP的錶達,流式細胞技術檢測各組細胞的凋亡.結果轉染TAP後的PC-3細胞中TAP錶達量明顯升高.RT-PCR結果顯示,H2O2單獨或聯閤促氧化劑丁胱亞磺酰亞胺(BSO)刺激細胞後TAP的mRNA錶達量分彆為(2.457±0.113)、(3.643±0.162)較對照組(0.721±0.048)顯著上升,差異有統計學意義(P<0.01);在無H2O2刺激的對照組PC-3、PC-3-vector、PC-3-TAP細胞的凋亡率分彆為2.42%、3.69%、15.37%;在H2O2刺激下,各組凋亡率分彆為7.56%、16.71%、43.93%;聯閤應用BSO與H2O2,各組凋亡分彆為13.73%、26.36%、49.19%.結論 TAP基因可參與前列腺癌細胞對氧化應激的應答反應,併促進氧化應激所緻的前列腺癌細胞凋亡.
목적 관찰생육분결합단백(TAP)대양화응격소치전렬선암세포조망적조공작용.방법 지질체법분별전염pEGFP-N3-TAP질립급공재체질립진입전렬선암세포PC-3.이과양화경(H2O2)자격PC-3세포구건양화응격모형,역전록-취합매련반응(RT-PCR)、Western blot검측TAP적표체,류식세포기술검측각조세포적조망.결과전염TAP후적PC-3세포중TAP표체량명현승고.RT-PCR결과현시,H2O2단독혹연합촉양화제정광아광선아알(BSO)자격세포후TAP적mRNA표체량분별위(2.457±0.113)、(3.643±0.162)교대조조(0.721±0.048)현저상승,차이유통계학의의(P<0.01);재무H2O2자격적대조조PC-3、PC-3-vector、PC-3-TAP세포적조망솔분별위2.42%、3.69%、15.37%;재H2O2자격하,각조조망솔분별위7.56%、16.71%、43.93%;연합응용BSO여H2O2,각조조망분별위13.73%、26.36%、49.19%.결론 TAP기인가삼여전렬선암세포대양화응격적응답반응,병촉진양화응격소치적전렬선암세포조망.
Objective To investigate the effect of α-tocopherol associated protein (TAP) gene in the regulation of the apoptosis induced by oxidative stress.Methods The plasmid pEGFP-N3-TAP expressing human TAP gene was transfected into PC-3 cells by the transfection reagent DMRIEC,and the expression of TAP was detected by Western blotting.Reverse transcription-polymerase chain reaction ( RT-PCR) and Western blotting were used to detect the change of TAP gene expression in the oxidative stress cell model established by using hydrogen peroxide ( H2O2 ).The apoptosis rate under oxidative stress in PC-3 cells was evaluated by the flow cytometry.Results After transfection,the expression level of TAP gene was significantly higher than the wild type PC-3 cell line.The TAP mRNA expression level in the groups of hydrogen peroxide alone (2.457 ± 0.113 ) or hydrogen peroxide combined with 250 μmol/L BSO (3.643 ± 0.162) was significantly higher than in control group (0.721 ± 0.048 ) ( P< 0.01 ).The flow cytometry suggested that in control group,the apoptosis rate in PC-3 cells,PC-3-TAP cells,PC-3-vector cells was 2.42%,3.69% and 15.37% respectively; In the hydrogen peroxide-stimulated groups,the apoptosis rate was 7.56%,16.71% and 43.93%,respectively; In the group of BSO combined with hydrogen peroxide,the apoptosis rate was 13.73%,26.36% and 49.19%,respectively.Conclusion TAP gene was involved in the response of oxidative stress in prostate cancer cells,and could promote the apoptosis induced by oxidative stress.
