三氧化二砷（As2O3）诱导乳腺癌细胞株MCF-7凋亡的实验研究  

Experimental Study on Apoptosis in Breast Cancer Cell Line MCF-7 Induced by Arsenic Trioxide

作　　者：张焕萍[1] 姜智[1] 范育利[1] 徐岚[1] 吴士良[1] 

机构地区：[1]苏州大学医学部生物化学与分子生物学系,江苏苏州215123

出　　处：《中国血液流变学杂志》2011年第3期384-388,共5页Chinese Journal of Hemorheology

基　　金：国家自然科学基金资助项目（30670462）

摘　　要：目的 观察不同浓度的三氧化二砷（As2O3）作用不同时间段后对乳腺癌细胞MCF-7的诱导凋亡作用和对β3GnT2 及β3GnT8表达的影响,并初步探讨其可能的作用机制.方法 不同浓度As2O3作用于乳腺癌细胞后,观察其对乳腺癌细胞生长状态的影响;用四甲基偶氮唑蓝（MTT）比色法观察其对MCF-7细胞凋亡的影响;同时采用RT-PCR 和Western Blot 检测相关基因mRNA及蛋白的表达水平变化.结果 不同浓度的As2O3 均可抑制乳腺癌细胞株MCF-7生长,其抑制作用随剂量增大而加强,呈时间剂量依赖性.RT-PCR 结果显示：β3GnT2、β3GnT8在各个浓度As2O3剂量组作用下,与空白组比较,均有不同程度的变化.1μg/mL和2μg/mL浓度组、4μg/mL和40μg/mL浓度组在相同时间段下分别比较,均有统计学差异（P ＜0.05）.Western Blot 检测相关蛋白结果与mRNA表达水平基本一致.β3GnT2蛋白表达中,4μg/mL（4h）组、40μg/mL（4h） 组分别与4μg/mL（48h）浓度组比较,均有明显差异（P＜0.05）;而在β3GnT8蛋白表达中,40μg/mL（4h）、4μg/mL（48h） 浓度组分别与空白组比较,有显著性差异（P＜0.05）.结论 As2O3对MCF-7细胞有显著的体外生长抑制作用,其抑制作用呈剂量依赖性;As2O3在体外能诱导MCF-7细胞凋亡,凋亡效应与剂量呈正相关;As2O3还能下调β3GnT2和β3GnT8基因的表达.Objective To observe the inhibition of As2O3 with different concentration on MCF-7 cell growth,the inducing apotosis function on the cell and the effect on β 3GnT2 and β 3GnT8 expression,and initially discuss the mechanism of As203 inducing apoptosis. Methods The effect of arsenic trioxide on the growth of breast cancer cell line MCF-7 was observed after applying arsenic trioxide of different concentrations in vitro. The inhibitory effect of arsenic trioxide on the cell proliferation was investigated with 3-（4,5-dimethylthizazol- 2-yl）-2,5-diphenyltetrazolium bromide（MTT）,reverse RT-PCR was adopted to detect mRNA horizontal express status of [3 3GnT2 and β 3GnT8;Western Blot was used to detect express status of related protein.Results As203 of different concentration could inhibit the growth of breast cancer cell line MCF-7,and its inhibition strengthened,along with the dose increasing,assuming the dose dependence.The mRNA show that there was a significant difference between any β 3GriT2 and β 3GriT8 group and the negative control;the comparison between 1 μg/mL and 2 μg/mL group,4 μg/mL and 40 μg/mL group the apotosis rates were obviously statistically different（P 〈 0.05）.The protein express status detected by Western Blot method was in accordance with the level of mRNA.In the expression of 13 3GnT2,there were obviously statistical difference among the 4 μg/ mL（4h） group,40 μg/mL（4h） group and 4 μg/mL（48h） group（P 〈 0.05）,but in the expression of [3 3GnT8,there was a significant difference between 40 μg/mL（4h） group and 4 μg/mL（48h）group and the negative control （P 〈 0.05）.Conclusion Arsenic trioxide can exert obvious inhibitory effect on MCF-7 cell growth in vitro.The inhibitory effect increases with dosage escalation and shows dosage dependence.As2O3 can induce the MCF-7 cell apoptosis,and the apoptosis effect and the dosage assume the positive correlation.Arsenic trioxide can regulate β 3GnT2 and β 3GriT8 expression down.

关 键 词：乳腺癌细胞株 三氧化二砷 凋亡 糖基转移酶 

分 类 号：R73-3[医药卫生—肿瘤]