摘要
目的:探讨顺铂(顺式二氨二氯铂)(c i sdichlorodiamineplatinum,DDP)对人结肠癌Caco-2细胞凋亡及相关蛋白Bcl-2、Caspase9、Caspase3表达的影响.方法:体外培养结肠癌Caco-2细胞;检测不同浓度DDP干预下MTT染色的A值,判定其对人结肠癌细胞恶性增殖的影响;不同浓度D D P对人结肠癌细胞凋亡的影响使用流式细胞仪进行检测,Western blot检测不同浓度DDP对人结肠癌细胞内Bcl-2蛋白的表达;应用分光光度法检测不同浓度DDP对人结肠癌细胞内Caspase9、Caspase3蛋白活性的影响.结果:(1)癌细胞增殖结果显示,在一定的作用时间范围内,Caco-2细胞存活率与DDP浓度呈负相关,具有剂量依赖性,其中4.000、2.000μg/m L干预的各组细胞抑制率最显著,差异均有统计学意义(P<0.05).而低剂量组0.250、0.125μg/m L及对照DMSO干预组细胞存活率比较差异均无统计学意义(P>0.05),其细胞存活率明显高于其他各浓度组,差异有统计学意义(P<0.01);(2)流式细胞仪检测结果显示:DDP能诱导Caco-2细胞的凋亡,其诱导凋亡的效果呈时间和剂量性依赖;(3)Western blot检测结果显示:Bcl-2蛋白在结肠癌Caco-2细胞中低表达,DDP干预Caco-2细胞72 h后,与对照组比较,Bcl-2蛋白表达明显下降(P<0.05).Caspase9、Caspase3在Caco-2细胞中低表达,DDP干预Caco-2细胞48、72 h后,与对照组比较,Caspase9、Caspase3表达明显升高(P<0.01).结论:顺铂可以抑制Caco-2细胞增殖、诱导Caco-2细胞凋亡,其机制可能与活化Caco-2细胞中Caspase9、Caspase3蛋白以及抑制Bcl-2蛋白表达有关.
AIM： To investigate whether cisplatin inducesthe apoptosis of colon cancer Caco-2 cells and inhibits the expression of Bcl-2, Caspase9 and Caspase3. METHODS： Caco-2 cells were treated with cisplatin. Cell proliferation was measured by MTT assay and fluorescence microscopy. The protein expression of Bcl-2 was determined by Western blot, and the protein expression of Caspase9 and Caspase3 was determined by spectrophotometry. RESULTS： Cisplatin inhibited the proliferation of Caco-2 cells in a concentration-dependent manner, and the survival rates of cells treated with 4 and 2 μg /mL cisplatin showed a statistical difference（P〈0.01）, although there was no significant difference between cells treated with l.0 and 0.5 μg/m L cisplatin（P〈0.05）. Cisplatin induced the apoptosis of Caco-2 cells in a time- and dose-dependent manner. Bcl-2 protein was highly expressed in Caco-2 cells. Cisplatin intervention for 72 h significantly reduced the expression of Bcl-2 protein in Caco-2 cells compared with control cells（P〉0.05）.The protein expression of Caspase9 and Caspase3 was significantly upregulated at 48 and 72 h after treatment with cisplatin when compared with control cells（P〈0.01）.CONCLUSION： Cisplatin inhibits the proliferation and induces apoptosis of Caco-2 cells via mechanisms that may be related to activation of Caspase9 and Caspase3 and inhibition of Bcl-2 protein expression.
出处
《世界华人消化杂志》
CAS
2015年第9期1460-1464,共5页
World Chinese Journal of Digestology
