熊果酸对人舌癌细胞增殖与凋亡的影响及其机制研究

Effect and mechanisms of ursolic acid on proliferation and apoptosis in human tongue cancer cells

目的探讨熊果酸(UA)对人舌癌Tca8113细胞体外增殖、凋亡的影响及其机制。方法体外培养Tca8113细胞并将其分为3组:空白组和低、高2个剂量实验组。空白组不做任何处理,实验组用低、高2个剂量(40,80μmol·L^-1)熊果酸处理24 h。以CCK-8法检测细胞增殖情况;以核酸荧光染料染色法和流式细胞术检测细胞凋亡情况;以蛋白质印迹法检测磷脂酰肌醇3-激酶(Pi3k)、磷酸化蛋白激酶B(p-Akt)和磷酸化细胞外调节蛋白激酶(p-Erk)和磷酸化P38丝裂原活化蛋白激酶(p-P38MAPK)蛋白表达水平。结果空白组和低、高2个剂量实验组的细胞存活率分别为(100.00±0.00)%,(76.27±5.54)%和(49.25±2.69)%;这3组的细胞凋亡率分别为(1.54±1.01)%,(20.79±1.96)%和(54.38±1.81)%;这3组的Pi3k蛋白的相对表达量分别为1.07±0.05,0.81±0.11和0.41±0.07;这3组的p-Akt蛋白的相对表达量分别为0.96±0.03,0.64±0.02和0.29±0.03;这3组的p-Erk蛋白的相对表达量分别为1.74±0.08,1.34±0.07和0.67±0.08;这3组的p-P38MAPK蛋白的相对表达量分别为0.87±0.03,1.17±0.06和1.46±0.09。上述指标:2个剂量实验组与空白组比较,差异均有统计学意义(均P<0.01)。结论熊果酸能显著抑制Tca8113细胞体外生长,其机制可能与下调Pi3k、p-Akt和p-Erk并上调p-P38MAPK的蛋白表达有关。

Objective To investigate the effect and mechanism of urso-lic acid on the proliferation and apoptosis of human tongue cancer Tca8113 cells in vitro.Methods Tca8113 cells were cultured in vitro and divided into three groups:blank group,low and high concentration experimental groups.The blank group was not treated.The experimental groups were treated with low and high concentrations(40,80μmol·L^-1)of ursolic acid for 24 h.Cell proliferation was detected by cell counting method.Apoptosis rates were detected by nucleic acid fluorescent dye(Hoechst33342)staining and flow cytometry.The expre-ssions level of proteins[phosphatidylinositol 3-kinase(Pi3 k),phosphorylated protein kinase B(p-Akt),phosphorylated extracellular regulates protein expression levels of protein kinase(p-Erk),phosphorylated P38 mitogen-activated protein kinase(p-P38 MAPK)]were detected by Western blotting.Results The cell survival rates in blank group and low,high concentration experimental groups were(100.00±0.00)%,(76.27±5.54)%and(49.25±2.69)%,respectively;the apoptosis rates in the three groups were(1.54±1.01)%,(20.79±1.96)%and(54.38±1.81)%;the relative expressions of Pi3 k protein in the three groups were 1.07±0.05,0.81±0.11 and 0.41±0.07,respectively;the relative expressions of p-Akt protein in the three groups were 0.96±0.03,0.64±0.02 and 0.29±0.03,respectively;the relative expressions of p-Erk protein in the three groups were1.74±0.08,1.34±0.07 and 0.67±0.08,respectively;the relative expressions of p-P38 MAPK protein in the three groups were 0.87±0.03,1.17±0.06 and 1.46±0.09,respectively.Comparison between experimental groups and blank group,the differences of the factors were significant(all P<0.01).Conclusion Ursolic acid can significantly inhibit the growth of Tca8113 cells in vitro,and the mechanism may be related to down-regulate Pi3 k,p-Akt,p-Erk and up-regulate the expression of p-P38 MAPK protein.