CAJ | 학술논문

目的初步探讨阿托伐他汀干预对恶性胶质瘤血管生成拟态的影响及作用机制.方法采用CCK-8实验检测梯度浓度阿托伐他汀(1010～104 mol/L)干预下胶质瘤细胞系U87细胞活性,以筛选出阿托伐他汀的药物安全浓度.将U87细胞置于含有阿托伐他汀不同安全浓度的细胞培养基及不含药的对照培养基中孵育,在基质胶Matrgel作用下进行三维培养以建立体外血管生成拟态模型,观察其血管生成拟态能力和计数其管腔样结构长度,应用RT-PCR、Western blotting实验分别检测药物干预组与对照组U87细胞中基质金属蛋白酶-2(MMP-2)mRNA及蛋白表达水平.结果 CCK-8实验筛选得到的阿托伐他汀的药物安全浓度为10-7、10-6和10-5 mol/L.U87细胞在接种后6～8h即可形成典型的网状管道血管生成拟态结构,且随阿托伐他汀药物浓度递增,细胞间形成的网状管道结构越来越少,同时10-7、10-6、10-5 mol/L阿托伐他汀组管腔样结构长度计数与对照组比较明显减少,MMP-2 mRNA及蛋白表达明显降低,差异均具有统计学意义(m0.05). 结论阿托伐他汀可能是通过抑制MMP-2表达而显著抑制U87细胞血管生成拟态的发生.
목적 초보탐토아탁벌타정간예대악성효질류혈관생성의태적영향급작용궤제.방법 채용CCK-8실험검측제도농도아탁벌타정(1010～104 mol/L)간예하효질류세포계U87세포활성,이사선출아탁벌타정적약물안전농도.장U87세포치우함유아탁벌타정불동안전농도적세포배양기급불함약적대조배양기중부육,재기질효Matrgel작용하진행삼유배양이건입체외혈관생성의태모형,관찰기혈관생성의태능력화계수기관강양결구장도,응용RT-PCR、Western blotting실험분별검측약물간예조여대조조U87세포중기질금속단백매-2(MMP-2)mRNA급단백표체수평.결과 CCK-8실험사선득도적아탁벌타정적약물안전농도위10-7、10-6화10-5 mol/L.U87세포재접충후6～8h즉가형성전형적망상관도혈관생성의태결구,차수아탁벌타정약물농도체증,세포간형성적망상관도결구월래월소,동시10-7、10-6、10-5 mol/L아탁벌타정조관강양결구장도계수여대조조비교명현감소,MMP-2 mRNA급단백표체명현강저,차이균구유통계학의의(m0.05). 결론 아탁벌타정가능시통과억제MMP-2표체이현저억제U87세포혈관생성의태적발생.
Objective To preliminarily explore the influence of intervention of atorvastatin in vasculogenic mimicry of glioblastoma and its mechanism.Methods CCK-8 experiment was carried out to detect the cell activity of glioma cell line U87 under the intervention of atorvastatin at different concentrations (1010-10-4 mol/L),aiming at screening out the safety concentration of atorvastatin.U87 cells were incubated in the mediums containing atorvastatin with different safety concentrations as well as in the control medium without atorvastatin.Three-D cultivation was performed in the Matrgel to establish the in vitro vasculogenic mimicry models,then the ability ofvasculogenic mimicry was observed and the length of the tube structure was counted.RT-PCR and Western blotting were performed to detect the mRNA and protein expression levels of matrix metalloproteinase-2 (MMP-2) in the U87 cells of intervention group and control group,respectively.Results Safety concentrations of atorvastatin screened by CCK-8 experiment were 10-7,10-6 and 10-5 mol/L.U87 cells formed the typical vasculogenic mimicry tube structure at 6-8 h after inoculation; the number of tube structure decreased with the increase ofatorvastatin concentration.Compared with the control group,the length counting of tube structure in intervention groups (10-7,10-6 and 10-5 mol/L) was significantly reduced and the mRNA and protein expression levels of MMP-2 were markedly decreased (P＜0.05).Conclusion Atovastatin significantly inhibits the vasculogenic mimicry formation via down-regulating the MMP-2 expression level.