小白菊内酯通过CDCA4调控PI3K/AKT通路抑制三阴性乳腺癌转移

Parthenolide inhibits the metastasis of Triple-Negative breast cancer by regulating CDCA4 and PI3K/AKT pathway

目的研究小白菊内酯(PTL)对乳腺癌MDA-MB-231细胞转移干预作用的影响及其相关机制。方法配置0、2.5、5.0、10.0、20.0、40.0μmol/L的PTL处理乳腺癌MDA-MB-231细胞24 h,采用MTS法和划痕实验检测不同浓度的PTL对MDA-MB-231细胞活力和迁移能力的影响。利用转染技术建立空白组,阴性对照组和CDCA4转染组;用PI3K抑制剂处理细胞24 h,建立PI3K抑制剂组和对照组。利用蛋白质印迹法和RT-PCR技术检测各组细胞人类细胞分裂周期相关基因4(CDCA4)、磷脂酰肌醇-3-激酶催化亚基α(PIK3CA)及丝氨酸/苏氨酸蛋白激酶(AKT)等信号分子的蛋白水平和mRNA的表达水平。结果PTL可抑制乳腺癌MDA-MB-231细胞的增殖和迁移,并且能够下调CDCA4、PIK3CA、AKT的蛋白和mRNA表达水平,差异均有统计学意义(P<0.05)。与空白组和阴性对照组相比,CDCA4转染后的PIK3CA、AKT的mRNA和蛋白的表达量明显减少,差异均有统计学意义(P<0.05)。与对照组相比,PI3K抑制剂组的AKT、CDCA4的蛋白表达量明显下降,差异均有统计学意义(P<0.05)。结论PTL可能通过下调CDCA4的表达,减少PIK3CA的突变从而抑制PI3K/AKT信号通路的激活,降低乳腺癌MDA-MB-231细胞的迁移能力。

Objective To investigate the effects of parthenolide(PTL)on the metastasis of breast cancer MDA-MB-231 cells and its related mechanisms.Methods Configure 0μmol/L,2.5μmol/L,5.0μmol/L,10.0μmol/L,20.0μmol/L,40.0μmol/L PTL was used to treat breast cancer MDA-MB-231 cells 24 h and the effects of different concentrations of PTL on the viability and migration of MDA-MB-231cells were detected by MTS method and scratch method.the blank control group,negative control group and CDCA4 plasmid transfection group were established by transfection technology.The cells were treated with PI3K inhibitor for 24 hours,and the model group and blank control group were established.Western blotting and RT-PCR were used to measure the expression levels of Human cell division cycle related gene 4(CDCA4),phosphatidylinositol-3-kinaseα(PIK3CA)and serine-threonine kinase(AKT)in each group.Results PTL inhibited the proliferation and migration ability of breast cancer MDA-MB-231 cells,and down regulated the expression of CDCA4,PIK3CA and AKT,the differences were statistically significant(P<0.05).The mRNA and protein expressions of PIK3CA and AKT in CDCA4 transfection group decreased significantly,the differences were statistically significant(P<0.05).Compared with the control group,the protein expressions of AKT and CDCA4 in the PI3K inhibitor group were significantly decreased,and the differences were statistically significant(P<0.05).Conclusion PTL may decrease the migration ability of MDA-MB-231 breast cancer cells by downregulating the expression of CDCA4,reducing the mutation of PI3KCA and inhibiting the activation of PI3K/AKT signaling pathway.