三氧化二砷对体外人小肠癌HIC细胞增殖的作用和机制

Effects and mechanism of arsenic trioxide on proliferation of HIC cells in vitro

目的探讨三氧化二砷(ATO)对体外人小肠癌HIC细胞增殖的作用及其机制。方法取处于对数生长期的人小肠癌HIC细胞,5个实验组分别用10、15、20、40及60μmol/L的ATO处理,用等量磷酸盐缓冲液(PBS)处理作为空白对照组,采用实时无标记细胞分析(RTCA)系统120 h连续监测各组人小肠癌HIC细胞的细胞指数(CI)值变化;取处于对数生长期的人小肠癌HIC细胞,4个实验组分别用10、15、20及40μmol/L ATO处理,等量PBS处理作为空白对照组,采用Western blot检测各组人小肠癌HIC细胞多克隆抗体糖原合成酶激酶-3(GSK-3β)、细胞周期蛋白B1(cyclin B1)、细胞周期蛋白D1(cyclin D1)及骨髓细胞瘤病毒癌基因(c-Myc)的表达。结果实验组经过10、15、20、40及60μmol/L ATO处理后的人小肠癌HIC细胞CI值均较空白对照组明显下降,其下降趋势随着ATO浓度的增加更明显;10、15、20及40μmol/L ATO组人小肠癌HIC细胞GSK-3β、cyclin B1、cyclin D1及c-Myc等相关周期蛋白的表达水平均较空白对照组均下降(P<0.05),且随着ATO浓度的增高,周期相关蛋白的表达水平越低。结论 ATO可抑制体外小肠癌小肠癌HIC细胞的增殖,其机制可能与相关周期蛋白cyclin B1、cyclin D1、GSK-3β及c-Myc的表达下调有关。

Objective To investigate the effect and mechanism of arsenic trioxide( ATO) on the proliferation of HIC cells in vitro. Methods The HIC cells of human small bowel cancer in logarithmic growth period were divided into 5 experimental groups: 10,15,20,40,and 60 μmol/L ATO. Equal phosphate buffer( PBS) was used as blank control group. The changes of cell index( CI) values of HIC cells in each group were monitored continuously by real-time label-free cell analysis( RTCA) system 120 h. The HIC cells of human small bowel cancer in logarithmic growth period were treated with 10,15,20,and 40 μmol/L ATO and treated with the same amount of PBS as the blank control group. The expression of glycogen synthase kinase-3( GSK-3β),cyclin B1( cyclin B1),cyclin D1( cyclin D1) and myeloma virus oncogene( c-Myc) were detected by Western blot.Results The cell CI of human small bowel cancer in 10,15,20,40,and 60 μmol/L ATO groups were significantly lower than those in the blank control group. As the ATO concentration increases,the decreasing trend is more obvious. The expression levels of HIC cell GSK-3β,cyclin B1,cyclin D1,and c-Myc in 10,15,20,and 40 μmol/L ATO groups were lower than those in the control group( P < 0. 05). As the concentration of ATO increased,the expression of cyclin decreased. Conclusion ATO may inhibit the proliferation of small intestinal cancer HIC cells in vitro,and its mechanism may be related to the down-regulation of related cyclin cyclin B1,cyclin D1,GSK-3β,and c-Myc expression.