基于c-Jun氨基端激酶/应激活化蛋白激酶信号通路探讨丹皮酚抑制结肠癌HCT116细胞生长的机制研究

Mechanism of paeonol inhibiting the growth of colon cancer HCT116 cells based on the c-Jun NH2-terminal kinase/stress-activated protein kinase signaling pathway

目的基于c-Jun氨基端激酶(JNK)/应激活化蛋白激酶(SAPK)信号通路探讨丹皮酚(PEON)抑制结肠癌HCT116细胞生长的机制。方法将结肠癌细胞株HCT116分为对照组和低、中、高剂量实验组。对照组仅给予不加任何药物的常规培养基进行培养;低、中、高剂量实验组分别给予含0.4,0.8和1.6 mmol·L^(-1)PEON的常规培养基进行培养。用噻唑蓝法检测HCT116细胞增殖能力的变化,用Hoechst33342/PI荧光染色法检测HCT116细胞的凋亡情况,用蛋白质印记法检测JNK、磷酸化JNK(p-JNK)和磷酸化细胞外信号传导激酶(p-ERK)蛋白的表达情况。结果低、高剂量实验组和对照组细胞的增殖抑制率分别为(36.69±1.33)%,(79.04±1.75)%和(17.21±1.19)%,凋亡率分别为(32.18±1.63)%,(58.56±2.23)%和(3.11±0.52)%,JNK相对表达量分别为0.72±0.03,0.35±0.02和1.01±0.04,p-JNK相对表达量分别为0.76±0.04,0.43±0.03和0.99±0.04,p-ERK相对表达量分别为0.58±0.04,0.27±0.03和1.03±0.03,低、高剂量实验组的上述指标与对照组相比,差异均有统计意义(均P<0.05)。结论PEON具有抑制结肠癌HCT116生长的作用,其机制可能是抑制JNK/SAPK信号通路,致使其下游基因蛋白表达下调。

Objective To investigate the inhibition mechanism of paeonol(PEON)on the growth of colon cancer HCT116 cells based on the c-Jun NH2-terminal kinase(JNK)/stress-activated protein kinase(SAPK)signaling pathway.Methods Colon cancer HCT116 cells were divided into control group and experimental-L,-M,-H groups.The control group was only given conventional culture medium without any drugs.The experimental-L,-M,-H groups were given conventional culture medium with 0.4,0.8 and 1.6 mmol·L^(-1)PEON,respectively.Methyl thiazolyl tetrazolium method was used to detect the proliferation of HCT116 cells.The apoptosis of HCT116 was detected by Hoechst33342/PI fluorescence staining.The Western blot was used to detect the expressions of JNK,phosphorylated JNK(p-JNK)and phosphorylated extracellular signal-regulated kinase(p-ERK)protein.Results The proliferation inhibition rates of experimental-L,-H groups and control group were(36.69±1.33)%,(79.04±1.75)%and(17.21±1.19)%,the cell apoptosis rates were(32.18±1.63)%,(58.56±2.23)%and(3.11±0.52)%,the relative expressions of JNK were 0.72±0.03,0.35±0.02 and 1.01±0.04,the relative expressions of p-JNK were 0.76±0.04,0.43±0.03 and 0.99±0.04,the relative expressions of p-ERK were 0.58±0.04,0.27±0.03 and 1.03±0.03.The above-mentioned indexes in the experimental-L,-H groups were significantly different from those in control group(all P<0.05).Conclusion PEON could inhibit the growth of colon cancer HCT116,and its mechanism may be that paeonol inhibits the JNK/SAPK signaling pathway,resulting in down-regulation of downstream gene protein expression.