摘要
目的探讨魟鱼骨素和环王巴明对人乳腺癌细胞系MCF-7是否具有协同作用,二者是否存在最佳配伍比例,以及解释该现象可能存在的分子生物学机制。方法分别将魟鱼骨素与环王巴明配制成10μmol/L浓度,按体积比1︰4、2︰3、1︰1、3︰2、4︰1进行配伍;采用四甲基偶氮唑盐法(MTT)检测各配伍组MCF-7细胞增殖情况;流式细胞术(FCM)检测各组药物作用前后细胞周期的变化及凋亡率;Western-blotting法检测细胞周期蛋白cyclin D1及P21蛋白表达的变化。结果 MTT提示1︰1配伍组对细胞增殖抑制作用明显优于其他药物组(P<0.05),差异具有统计学意义;FCM法提示1︰1配伍组作用细胞24 h与单个用药作用48 h所得结果差异无统计学意义(P>0.05);Western-blotting提示1︰1配伍组作用细胞24 h与单个用药作用48 h细胞周期蛋白cyclin D1与P21灰度值差异无统计学意义(P>0.05)。结论魟鱼骨素和环王巴明存在协同作用及最佳药物配伍比例(两药浓度分别为10μmol/L,两药按体积比1︰1配伍),24 h可达到最佳作用效果,其分子生物学机制与cyclin D1失表达、P21过表达有关。
【Objective】 To investigate whether ray cartilage extract（RCE） combined with cyclopamine have cooperation effect on proliferation of human mammary carcinoma cell line MCF-7 or not and whether they have the best compound proportion or not and to explain its possible molecule biological mechanism of the phenomenon.【Methods】 To make up the concentration of RCE and cyclopamine to 10 μmol/L and then to compound the two drug according to volume proportion of 1：4,2：3,1：1,3：2,4：1.Methabenzthiazuron（MTT） assay was used to evaluate the proliferation of the cells.The expressions of the cyclin D1 and p21 in MCF-7 cells were determined by western blotting.【Results】 MTT pointed out the drug group of 1：1 had obvious effect compared with other drug groups and the difference had statistics significance（P〈0.05）.FCM pointed out that the difference of the results between the drug group of 1：1 effecting on cells for 24 hours and each single drug group effecting on cells for 48 hours have no statistics significance（P〈0.05）.Western blotting pointed out that the difference of the gray value of cyclin D1 and p21 between the drug group of 1：1 effecting on cells for 24 hours and the each drug group effecting on cells for 48 hours have no statistics significance（P〈0.05）.【Conclusions】 RCE and cyclopamine have cooperation effect and the best compound proportion is to compound the two drugs which concentration are 10 μmol/L according to volume proportion of 1：1.And the drug group of 1：1 has the best effect time of 24 hours.And the molecule biological mechanism may be related to the lack-expression of the cyclin D1 and the over-expression of the p21.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2012年第29期26-30,共5页
China Journal of Modern Medicine
基金
辽宁省教育厅高等学校科研项目(No:2008398)
