蛇莓对人结肠癌RKO细胞失巢凋亡作用的实验研究

目的观察蛇莓对人结肠癌RKO细胞悬浮生长及失巢凋亡作用。方法不同浓度蛇莓作用RKO细胞,CCK-8（Cell Counting Kit-8,CCK-8）法检测蛇莓对RKO细胞悬浮生长作用,双嵌入剂乙锭均二聚物（Ethidium homodimer-1,EthD-1）染色检测失巢凋亡,比色法检测半胱天冬酶-3（Caspase-3）活性,二氯氢化荧光素二酯染色法结合荧光酶标仪检测细胞内活性氧的产生。结果蛇莓可显著抑制RKO悬浮生长,呈剂量依赖性。蛇莓作用后悬浮生长RKO细胞吸收EthD-1散发红色荧光,部分细胞可见核碎裂,呈现凋亡形态改变;蛇莓同时可显著增强RKO细胞Caspase-3活性,升高细胞内活性氧水平。结论蛇莓可抑制RKO细胞悬浮生长,促使RKO细胞发生失巢凋亡,可能与活化Caspase-3以及升高细胞内活性氧水平相关。

Experimental Research of RKO Cell Anoikis Induced by Duchesnea Indica in Human Colon Cancer

Objective To observe RKO cell suspending growth and anoikis induced by duchesnea indica in human colon cancer.Methods Duchesnea indica of different concentrations worked on RKO cell.CCK-8（Cell Counting Kit-8）method was used to detect the impact of duchesnea indica on RKO cell suspending growth.Ethidium homodimer-1（EthD-1）staining was adopted to detect anoikis.The colorimetric method was applied to detect caspase-3 activity.The dichlorohyrogenated fluorescein diester staining method and fluorescence microplate system were used to co-detect the production of reactive oxygen in the cell.The fluorescent probe DCFH-DA was used to detect the reactive oxygen.Results Duchesnea indica inhibited significantly RKO cell suspending growth,presenting dose dependence.After stimulated with duchesnea indica,RKO in suspending growth absorbed EthD-1 and emitted red fluorescence.The nuclear fragmentation was seen in a part of cells,presenting apoptotic morphological change.Duchesnea indica enhanced significantly caspase-3 activity in RKO cell and increased the level of reactive oxygen.Conclusion Duchesnea indica inhibits the suspending growth of RKO cell,promotes anoikis in RKO cell,which is probably related to the activation of caspase-3 and the increase of intracellular reactive oxygen level.