肌醇六磷酸诱导HT-29细胞凋亡及相关基因表达
Effect of inositol hexaphosphate on apoptosis of HT-29 human colon carcinoma cells and expression of apoptosis related gene
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摘要: 目的 研究肌醇六磷酸(IP6)对人结肠癌细胞系(HT-29)的凋亡诱导作用.方法 将不同浓度IP6以不同时间作用于HT-29细胞,运用透射电镜观察IP6对HT-29细胞凋亡作用;流式细胞仪检测细胞凋亡率变化;钙离子荧光探针(Fura-2/AM)法测定细胞内Ca2+浓度变化;RT-PCR法检测细胞内caspase-3 mRNA表达.结果 IP6诱导HT-29细胞发生凋亡的形态学变化为染色体固缩,可见凋亡小体;3.3mmol/L IP6作用6个月HT-29细胞凋亡率为4.4%,细胞内Ca2+浓度为38.66 mmol/L,与对照组比较明显升高(P<0.05);各IP6组Caspase-3mRNA表达均升高(P<0.05);IP6诱导HT-29细胞凋亡作用及Ca2+、Caspase-3mRNA的表达变化具有时间依赖关系.结论 IP6对HT-29细胞有诱导凋亡作用,其机制可能是通过调节细胞内Ca2+浓度及Caspase-3的表达来实现.Abstract: Objective To investigate effect and mechanism of inositol hexaphosphate(IP6)on HT-29 human colon carcinoma cell line.Methods HT-29 cells were exposed to various concentrations of IP6 for differenttine The effect of apoptosis was detected by tmnsm fission electron microscopy(TFM).Flowcytometric analysis was perfouned to detect cell apoptosis Fura-2/AM was used to deteun fine the concentration of intracellular-free calcium.Caspase3 mRNA expression in cells was deteun fined by reverse transcription polymerase chain reaction(RT-PCR).Results Moiphological exam ination showed cells shrinkage and the presence of "apoptopic bodies".IP6 increased the rate of cell apoptosis with a tine-dependent trend(P < 0.05),the concentration of intmcellularfree calcium(P<0.05),and the expression of Caspase3 mRNA(P < 0.05).Conclusion IP6 could induce HT-29 cell apoptosis Its mechanism may relates to the regulation of intracellularfree calcium and the expression of apoptosis related gene
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