熊果苷抑制胰腺癌细胞生长及好氧糖酵解的研究

Research of arbutin inhibiting pancreatic cancer cell growth and aerobic glycolysis

目的研究熊果苷对胰腺癌细胞生长及好氧糖酵解的影响及机制。方法将细胞分为对照组、阳性对照组和低、中、高剂量实验组、转染组和高剂量实验-转染组。对照组给予等量蒸馏水处置;阳性对照组给予10μmol·L^-1顺铂处置;低、中、高剂量实验组分别给予5,10,20μmol·L^-1熊果苷处置;转染组给予转染sh-HIPK2质粒处置;高剂量实验-转染组转染sh-HIPK2质粒后,给予20μmol·L^-1熊果苷处置。用克隆形成实验检测细胞生长,用流式细胞仪检测细胞凋亡率,用Seahorse XFp细胞代谢分析仪检测好氧糖酵解水平。结果低、高剂量实验组和对照组、转染组和高剂量实验-转染组的细胞克隆数目分别为(102.68±5.48),(65.99±2.60),(178.79±4.30),(254.00±4.39)和(184.81±3.63)个,细胞凋亡率分别为(26.78±0.06)%,(42.31±0.06)%,(4.23±0.05)%,(1.23±0.03)%和(4.15±0.02)%,细胞外酸化率分别为(223.37±2.89),(185.28±1.96),(355.22±2.20),(424.66±2.90)和(365.78±3.56)mpH·min^-1,氧消耗率分别为(483.61±3.46),(588.06±4.82),(300.45±4.18),(221.69±4.17)和(287.33±4.18)pMoles·min^-1。低、高剂量实验组的上述指标与对照组比较,差异均有统计学意义(P<0.05,P<0.01)。高剂量实验-转染组的上述指标与高剂量实验组比较,差异均有统计学意义(均P<0.01)。结论熊果苷通过同源结构域相互作用的蛋白激酶2调节心肌肌球蛋白结合蛋白C的表达,从而抑制胰腺癌细胞生长及好氧糖酵解。

Objective To investigate the effect and mechanism of arbutin on the growth and aerobic glycolysis of pancreatic cancer cells.Methods The cells were divided into control,positive control,experimental-L,-M,-H groups,transfection group and experimental-H+transfection group.The control group was given the same amount of distilled water,positive control group was given 10μmol·L^-1 cisplatin,experimental-L,-M,-H groups were given arbutin 5,10,20μmol·L^-1,respectively,transfection group was treated with sh-HIPK2 transfection plasmid,experimental-H+transfection group was transfected with sh-HIPK2 plasmid and treated with arbutin 20μmol·L^-1.Cell growth was detected by colony formation assays,the rate of apoptosis detected by flow cytometry,the aerobic glycolysis level was analyzed by Seahorse XFp cell metabolism.Results The number of cell clones in experimental-L,-H groups,control group,transfection group and experimental-H+transfection group was(102.68±5.48),(65.99±2.60),(178.79±4.30),(254.00±4.39)and(184.81±3.63),the cell apoptosis rates were(26.78±0.06)%,(42.31±0.06)%,(4.23±0.05)%,(1.23±0.03)%and(4.15±0.02)%,the extracellular acidification rates were(223.37±2.89),(185.28±1.96),(355.22±2.20),(424.66±2.90)and(365.78±3.56)mp H·min^-1,and the oxygen consumption rates were(483.61±3.46),(588.06±4.82),(300.45±4.18),(221.69±4.17)and(287.33±4.18)p Moles·min^-1.Comparing with the control group,the differences of above indexes in the experimental-L,-H groups were statistically significant(P<0.05,P<0.01).Comparing with the experimental-H group,the above indexes in the experimental-H+transfection group had statistically significant differences(all P<0.01).Conclusion Arbutin inhibited the growth and aerobic glycolysis of pancreatic cancer cells through homologous domain interacting protein kinase 2 and regulated the expression of cardiac myosin binding protein C.