中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2012年
1期
45-49
,共5页
张月%张斌%冯炜红%李媛媛%曹旭晨
張月%張斌%馮煒紅%李媛媛%曹旭晨
장월%장빈%풍위홍%리원원%조욱신
乳腺肿瘤%细胞增殖%细胞凋亡
乳腺腫瘤%細胞增殖%細胞凋亡
유선종류%세포증식%세포조망
Breast neoplasms%Cell proliferation%Apoptosis
目的 研究Aurora激酶抑制剂PHA739358抑制人乳腺癌T47D细胞增殖、诱导凋亡的分子机制.方法 用四甲基偶氮唑盐法(MTT法)评价PHA739358对T47D细胞增殖的影响；免疫荧光法观察染色体、核的变化；流式细胞术检测细胞周期阻滞和凋亡率；Western印迹检测AuroraA、pAurora A、Histone H3、p-Histone H3,周期特异性指标Cyclin B1,周期调节性指标Cdc2、Cdc25c、p-Cdc2、p-Cdc25c,凋亡诱导指标p21、p53,凋亡相关指标PARP、Bcl-2、Bax.结果 不同浓度的PHA739358处理细胞24h、48 h后,明显抑制T47D的增殖,IC50分别为(3.44±0.54) μ,mol/L、(0.21±0.67)μmol/L,核与纺锤体形态发生明显变化,G2/M期阻滞增加呈剂量依赖性,Western印迹显示Aurora A、Histone H3、Cdc2、Cdc25c无明显趋势变化,p-AuroraA、p-Histone H3、CyclinB1、Bcl-2随着浓度的增加而减少,p-Cdc2、p-Cdc25c、P21、P53、Bax、PARP随着浓度的增加而增加.流式细胞术凋亡率由0.31％ ±0.03％增加到40.6％±0.81％.结论 PHA739358抑制乳腺癌细胞T47D增殖、诱导凋亡的分子机制为乳腺癌治疗提供了新思路.
目的 研究Aurora激酶抑製劑PHA739358抑製人乳腺癌T47D細胞增殖、誘導凋亡的分子機製.方法 用四甲基偶氮唑鹽法(MTT法)評價PHA739358對T47D細胞增殖的影響；免疫熒光法觀察染色體、覈的變化；流式細胞術檢測細胞週期阻滯和凋亡率；Western印跡檢測AuroraA、pAurora A、Histone H3、p-Histone H3,週期特異性指標Cyclin B1,週期調節性指標Cdc2、Cdc25c、p-Cdc2、p-Cdc25c,凋亡誘導指標p21、p53,凋亡相關指標PARP、Bcl-2、Bax.結果 不同濃度的PHA739358處理細胞24h、48 h後,明顯抑製T47D的增殖,IC50分彆為(3.44±0.54) μ,mol/L、(0.21±0.67)μmol/L,覈與紡錘體形態髮生明顯變化,G2/M期阻滯增加呈劑量依賴性,Western印跡顯示Aurora A、Histone H3、Cdc2、Cdc25c無明顯趨勢變化,p-AuroraA、p-Histone H3、CyclinB1、Bcl-2隨著濃度的增加而減少,p-Cdc2、p-Cdc25c、P21、P53、Bax、PARP隨著濃度的增加而增加.流式細胞術凋亡率由0.31％ ±0.03％增加到40.6％±0.81％.結論 PHA739358抑製乳腺癌細胞T47D增殖、誘導凋亡的分子機製為乳腺癌治療提供瞭新思路.
목적 연구Aurora격매억제제PHA739358억제인유선암T47D세포증식、유도조망적분자궤제.방법 용사갑기우담서염법(MTT법)평개PHA739358대T47D세포증식적영향；면역형광법관찰염색체、핵적변화；류식세포술검측세포주기조체화조망솔；Western인적검측AuroraA、pAurora A、Histone H3、p-Histone H3,주기특이성지표Cyclin B1,주기조절성지표Cdc2、Cdc25c、p-Cdc2、p-Cdc25c,조망유도지표p21、p53,조망상관지표PARP、Bcl-2、Bax.결과 불동농도적PHA739358처리세포24h、48 h후,명현억제T47D적증식,IC50분별위(3.44±0.54) μ,mol/L、(0.21±0.67)μmol/L,핵여방추체형태발생명현변화,G2/M기조체증가정제량의뢰성,Western인적현시Aurora A、Histone H3、Cdc2、Cdc25c무명현추세변화,p-AuroraA、p-Histone H3、CyclinB1、Bcl-2수착농도적증가이감소,p-Cdc2、p-Cdc25c、P21、P53、Bax、PARP수착농도적증가이증가.류식세포술조망솔유0.31％ ±0.03％증가도40.6％±0.81％.결론 PHA739358억제유선암세포T47D증식、유도조망적분자궤제위유선암치료제공료신사로.
Objective To explore the molecular mechanisms of Aurora kinase inhibitor PHA739358 in inhibited proliferation and in vitro induced apoptosis of breast cancer cells. Methods The in vitro cultured T47D cells in logarithmic growth phase were used.The effects of PHA739358 on cell proliferation were examined by MTT (methyl thiazolyl tetrazolium ) assay. The variety of nuclear and spindle morphologies was examined by immunofluorescence.And G2/M arrest was determined by flow cytometry.The morphological changes of apoptotic cells were observed by fluorescent microscope.The levels of Aurora kinase relative protein expression phosphorate-AuroraA (p-AuroraA),AuroraA,phosphorate-histone H3 (p-histone H3 ),histone H3,cell cycle-specific protein expression CyclinBl,cell cycle regulative protein expression Cdc2,Cdc25c,phosphorate-Cdc2 (p-Cdc2),phosphorate-Cdc25c (p-Cdc25c) and apoptosis relative protein expression PARP,Bcl-2 and Bax were detected by Western blot.The apoptotic rate was examined by flow cytometer.Results PHA739358 obviously inhibited the proliferation of T47D cells after a 24 h or 48 h treatment in a dose-dependent and time-dependent manner.Their IC50 values were (3.44:0.54) and ( 0.21 ± 0.67 ) μmol/L respectively. Flow cytometry showed that G2/M arrested in a dosedependent manner.PHA739358 dose-dependently and time-dependently promoted the dissection of PARP (poly (ADP-ribose) polymerase); down-regulated the expressions of Bcl-2,p-AuroraA,p-histone H3,CyclinB1 and up-regulated the levels of Bax,p-Cdc25c,p-Cdc2,P21 and P53 protein.PHA739358 had no significant effects on the expressions of AuroraA and histone H3. Flow cytometry and fluorescence microscope showed that PHA739358 significantly induced apoptosis. Flow cytometry showed the rate of apoptosis significantly increased from 0.31％ ± 0.03％ to 40.6％ ± 0.81％. Conclusion The proliferation-inhibiting and apoptosis-inducing effects of PHA739358 may provide new therapeutic approaches of breast cancer.
