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目的:研究甜菜红苷对人结肠癌HT29细胞凋亡的影响。方法:体外传代培养HT29细胞。以不同质量浓度[0(空白对照)、0.5、1、1.5、2、3 mg/ml]的甜菜红苷培养细胞24 h,以1.5 mg/ml甜菜红苷培养细胞不同时间[0(空白对照)、1、3、6、12、24 h]后,采用MTT法测定细胞活力并计算抑制率;以0(空白对照)、1、1.5、2 mg/ml甜菜红苷培养细胞24 h后,采用流式细胞仪测定细胞凋亡率,测定含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)、Caspase-9的活性,采用实时荧光聚合酶链反应(RT-PCR)与Western blot法测定Bcl-2、Bax m RNA与蛋白的表达。结果:与空白对照比较,0.5、1、1.5、2、3 mg/ml甜菜红苷培养细胞24 h后,1.5 mg/ml甜菜红苷培养细胞1、3、6、12、24 h后,细胞抑制率升高(P<0.01)。与空白对照比较,1、1.5、2 mg/ml甜菜红苷培养细胞24 h后,细胞凋亡率升高,Caspase-3、Caspase-9活性增强,Bcl-2 m RNA与蛋白表达减弱、Bax m RNA与蛋白表达增强(P<0.01)。结论:甜菜红苷可诱导HT29细胞凋亡,其机制可能与上调Caspase-3、Caspase-9活性和Bax m RNA与蛋白表达,下调Bcl-2 m RNA与蛋白表达有关。
Objective: To study the effects of betanin on the apoptosis of human colon cancer HT29 cells. Methods: HT29 cells were subcultured in vitro. The cells were incubated with beetins at different concentrations [0 (blank control), 0.5, 1, 1.5, 2, and 3 mg / ml] for 24 h and incubated with 1.5 mg / ), 1, 3, 6, 12 and 24 h], cell viability was measured by MTT method and the inhibition rate was calculated. Cells were cultured with 0 (blank control), 1,1.5,2 mg / ml beet glycoside for 24 h, The apoptotic rate was determined by flow cytometry. Caspase-3 and Caspase-9 activity were determined by real-time fluorescence polymerase chain reaction (RT-PCR) Western blot was used to detect the expression of Bcl-2, Bax m RNA and protein. RESULTS: Compared with the blank control, cells were cultured with 0.5,1,1.5,2,3 mg / ml beet glycoside for 24 h. Cells were cultured with 1.5 mg / ml beet glycoside for 1, 3, 6, 12 and 24 h. The inhibition rate increased (P <0.01). Compared with the blank control group, the cell apoptosis rate increased, the activity of Caspase-3 and Caspase-9 increased, the expression of Bcl-2 mRNA and protein decreased, Bax m RNA and protein expression increased (P <0.01). CONCLUSION: Beetoside can induce the apoptosis of HT29 cells. The mechanism may be related to the up-regulation of Caspase-3, Caspase-9 activity, Bax m RNA and protein expression, down-regulation of Bcl-2 mRNA and protein expression.