Aurora激酶抑制剂对乳腺癌细胞增殖和凋亡的影响  

Effects of aurora kinase inhibitor on proliferation and apoptosis of breast cancer cells

作　　者：张月[1] 张斌[1] 冯炜红[1] 李媛媛[1] 刘博文[1] 曹旭晨[1] 

机构地区：[1]天津医科大学附属肿瘤医院乳腺一科乳腺癌防治教育部重点实验室天津市“肿瘤防治”重点实验室,300060

出　　处：《中华实验外科杂志》2012年第1期74-76,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81001186）;天津市自然科学基金资助项目（IOJCYBJC14100）

摘　　要：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，用噻唑蓝（M1Tr）比色法和克隆形成实验检测乳腺癌细胞增殖，碘化丙锭（PI）单染法榆测细胞周期，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、48h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；克隆形成率由（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—MB-231增殖、诱导凋亡呈剂量依赖性。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 （MTr） 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 ） μmol/L. After treatment with VX-680, apoptosis of MDA-MB-231 ceils 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 ceils 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 phos-phorylated 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.

关 键 词：乳腺癌 VX-680 AURORA 增殖 脱噬作用 

分 类 号：R737[医药卫生—肿瘤]