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目的探讨槲皮素在体外对人结肠癌HT-29细胞凋亡的影响,并对其诱导机制进行研究。方法体外常规培养HT-29细胞,随机设定对照组和高、中、低剂量(32,16,8μg·m L-1)槲皮素组,药物干预24 h后,采用CCK-8试剂盒检测槲皮素对HT-29细胞增殖的影响,倒置显微镜观察细胞形态变化,蛋白免疫印迹法检测MKK3/6/p38信号通路蛋白表达,并同时检测凋亡相关蛋白Caspase 8、Caspase 3、Bax及Bcl-2的表达水平。结果槲皮素干扰HT-29细胞24 h后,CCK-8实验发现槲皮素可明显抑制细胞的增殖;形态学观察发现细胞密度明显减小、细胞体积变小;免疫印迹实验发现槲皮素可诱导MKK3/6及p38的磷酸化,进而促进促凋亡蛋白Caspase8、Caspase3及Bax的表达,抑制了抗凋亡蛋白Bcl-2的表达。结论槲皮素可诱导人结肠癌HT-29细胞凋亡,其作用机制可能与激活MKK3/6/p38信号通路有关。
Objective To investigate the effect of quercetin on the apoptosis of human colon cancer HT-29 cells in vitro and its mechanism of induction. Methods HT-29 cells were cultured in vitro. The control group and the quercetin groups at high, medium and low doses (32, 16, and 8μg · m L -1) were randomly assigned. After 24 h treatment with the drug, CCK-8 kit The effect of quercetin on the proliferation of HT-29 cells was observed. The morphological changes of cells were observed by inverted microscope. The expression of MKK3 / 6 / p38 signal pathway protein was detected by Western blotting. Caspase 8, Caspase 3, Bax and Bcl-2 expression levels. Results After treated with quercetin for 24 h, the proliferation of HT-29 cells was significantly inhibited by quercetin after 24 h treatment with CCK-8. The morphological observation showed that the cell density decreased and the cell volume decreased. The results of immunoblotting showed that quercetin Can induce phosphorylation of MKK3 / 6 and p38, further promote the expression of proapoptotic proteins Caspase8, Caspase3 and Bax, and inhibit the expression of anti-apoptotic protein Bcl-2. Conclusion Quercetin induces apoptosis in human colon cancer HT-29 cells, and its mechanism may be related to the activation of MKK3 / 6 / p38 signaling pathway.