CAJ | 학술논문

目的观察Aurora激酶抑制剂VX-680对人乳腺癌MDA-MB-231细胞株增殖和凋亡的影响.方法不同浓度的VX-680处理MDA-MB-231细胞,倒置显微镜下观察细胞形态变化,用噻唑监(MTT)比色法和克隆形成实验检测乳腺癌细胞增殖,碘化丙锭(PI)单染法检测细胞周期,免疫荧光观察核和纺锤体形态变化,免疫印迹法检测AuroraA/B蛋白、组蛋白和凋亡相关蛋白表达,Yo-Pro-1荧光染色观察细胞凋亡形态学变化.结果 VX-680显著抑制MDA-MB-231增殖,24、48 h半数抑制浓度( IC50)分别为(2.362±0.599)、(0.102±0.556) μmol/L;经药物作用24h后,贴壁细胞出现皱缩、变圆、脱落；克隆形成率南( 93.00 ±0.03)％降至(0.01±0.01)％,G0/G1期细胞由(51.27±0.75)％降至(5.87±0.49)％,G2/M期细胞由(17.67±1.25)％升高至(91.93±1.96)％,荧光染料法显示凋亡细胞数由(4.33±0.03)％增高至(44.00±0.04)％,实验组与对照组比较差异均有统计学意义(P＜0.05)；细胞核与纺锤体结构紊乱；免疫印迹检测提示磷酸化AuroraA/B、磷酸化Histone H3表达水平降低,Caspase-3和PARP被剪切.结论 VX-680抑制乳腺癌细胞MDA-MB231增殖、诱导凋亡呈剂量依赖性.
목적 관찰Aurora격매억제제VX-680대인유선암MDA-MB-231세포주증식화조망적영향.방법 불동농도적VX-680처리MDA-MB-231세포,도치현미경하관찰세포형태변화,용새서감(MTT)비색법화극륭형성실험검측유선암세포증식,전화병정(PI)단염법검측세포주기,면역형광관찰핵화방추체형태변화,면역인적법검측AuroraA/B단백、조단백화조망상관단백표체,Yo-Pro-1형광염색관찰세포조망형태학변화.결과 VX-680현저억제MDA-MB-231증식,24、48 h반수억제농도( IC50)분별위(2.362±0.599)、(0.102±0.556) μmol/L;경약물작용24h후,첩벽세포출현추축、변원、탈락；극륭형성솔남( 93.00 ±0.03)％강지(0.01±0.01)％,G0/G1기세포유(51.27±0.75)％강지(5.87±0.49)％,G2/M기세포유(17.67±1.25)％승고지(91.93±1.96)％,형광염료법현시조망세포수유(4.33±0.03)％증고지(44.00±0.04)％,실험조여대조조비교차이균유통계학의의(P＜0.05)；세포핵여방추체결구문란；면역인적검측제시린산화AuroraA/B、린산화Histone H3표체수평강저,Caspase-3화PARP피전절.결론 VX-680억제유선암세포MDA-MB231증식、유도조망정제량의뢰성.
Objective To investigate the effects of VX-680,a new aurora kinase inhibitor,on proliferation and apoptosis of MDA-MB-231 breast cancer cell lines.Methods MDA-MB-231 cells were cultured for 24 h in the medium which contained VX-680 with different concentrations.Inverted Microsoft showed cells varied with different concentrations.The effect of VX-680 on cell proliferation was examined by methyl thiazol tetrazolium (MTT) assay and colony assay.The cell cycle was analyzed by propidium iodide (PI) dyeing.The variety of nucleus and spindle morphology was tested by immunofluorescence.The levels of AuroraA/B protein,histone H3 and apoptosis relative protein expression were detected by Western blotting.Morphological changes of apoptotic cells were observed under the flurescent microscopy.Results VX-680 obviously inhibited the proliferation of MDA-MB-231 cells 24 h or 48 h after treatment,with the 50％ inhibitory concentration ( IC50 ) value being (2.362 ±0.599) μmol/L or (0.102 ±0.556) μnol/L.After treatment with VX-680,apoptosis of MDA-MB-231 cells was seen.The ratio of colony forming was decreased from (93.00 ±0.03)％ to (0.01 ±0.01 )％,the ratio of G0/G1 was decreased from (51.27 ±0.75)％ to (5.87 ±0.49)％,the percentage of G2/M cells was increased from (17.67 ± 1.25)％ to (91.93 ± 1.96)％,and the percentage of apoptotic cells was increased from (4.33 ±0.03)％ to (44.00 ±0.04 ) ％ ( P ＜ 0.05 ).The morphology of nucleus and spindle was abnormal.The expression levels of phosphorylated AuroraA/B and Histone H3 were significantly down-regulated,and the cleavage of PARP and Caspase-3 was detected by Western blotting.Conclusion VX-680 inhibited the proliferation and induced the apoptosis of MDA-MB-231 cells in a dose-dependent manner.