摘要
目的通过观察AQP1在黄芩苷调控肺腺癌细胞增殖、迁移、凋亡中的作用探讨其分子机制。方法将人肺腺癌细胞株LTEP-A2分为对照组及黄芩苷组,对照组正常培养细胞,黄芩苷组采用黄芩苷处理48h。采用qRT-PCR法检测细胞中AQP1表达,以CCK-8法、划痕实验、流式细胞术分别观察细胞增殖、迁移能力及凋亡情况。将LTEP-A2细胞分为对照组、AQP1降表达组、AQP1过表达组,对照组不做处理,AQP1降表达组、AQP1过表达组分别转染AQP1小干扰siRNA及AQP1过表达载体,按照上法观察细胞增殖、迁移能力及凋亡情况。将转染AQP1过表达载体的AQP1过表达LTEP-A2细胞分为AQP1过表达组、AQP1过表达+黄芩苷组、AQP1过表达+LY294002组、AQP1过表达+wortmannin组,AQP1过表达组不做处理,余下各组分别采用黄芩苷、PI3Kα抑制剂LY294002、磷酸化PI3Kα(p-PI3Kα)抑制剂wortmannin处理;采用Western blotting法观察各组PI3K/Akt信号通路相关蛋白血管内皮生长因子(VEGF)、胰岛素样生长因子(IGF)、哺乳动物雷帕霉素靶蛋白(mTOR)、苏氨酸激酶(p-Akt)、缺氧诱导因子1α(HIF-1α)表达。结果黄芩苷组AQP1表达、细胞增殖抑制率低于对照组,细胞迁移距离、细胞凋亡率高于对照组(P均<0.05)。细胞增殖抑制率AQP1过表达组>对照组>AQP1降表达组,细胞迁移距离、细胞凋亡率AQP1降表达组>对照组>AQP1过表达组(P均<0.05)。LTEP-A2细胞VEGF、IGF、mTOR、p-Akt、HIF-1α蛋白表达AQP1过表达组>AQP1过表达+黄芩苷组、AQP1过表达+LY294002组、AQP1过表达+wortmannin组(P均<0.05),过表达+黄芩苷组、AQP1过表达+LY294002组、AQP1过表达+wortmannin组各项蛋白表达差异均无统计学意义。结论黄芩苷通过下调AQP1表达抑制肺腺癌细胞的增殖、迁移并诱导其凋亡,其机制可能与抑制PI3K/Akt信号通路激活有关。
Objective To investigate the effects and mechanism of baicalin on the proliferation,migration,and apoptosis of lung adenocarcinoma cells through AQP1.Methods Human lung adenocarcinoma cell line LTEP-A2 was divided into the control group and baicalin group.Cells were cultured normally in the control group,and cells in the baicalin group were treated with baicalin for 48h.AQP1 expression was detected by qRT-PCR,and cell proliferation,migration,and apoptosis were observed by CCK-8,Scratch assay and flow cytometry,respectively.LTEP-A2 cells were divided into the control group,AQP1 down-expression group and AQP1 over-expression group.Cells in the control group were not treated,and cells in the AQP1 down-expression group and AQP1 over-expression group were transfected with small interfering siRNA of AQP1 and AQP1 over-expression vector,respectively.Cell proliferation,migration ability and apoptosis were observed according to the above method.AQP1 overexpression LTEP-A2 cells transfected with AQP1 over-expression vector were divided into the AQP1 over-expression group,AQP1 over-expression+baicalin group,AQP1 over-expression+LY294002 group,AQP1 over-expression+wortmannin group,respectively;cells in the AQP1 overexpression group were not treated,cells in the remaining groups were treated with baicalin,PI3Kα inhibitor LY294002,and phosphorylated PI3Kα(p-PI3Kα)inhibitor wortmannin,respectively.The expression levels of PI3K/Akt signaling pathway-related proteins vascular endothelial growth factor(VEGF),insulin-like growth factor(IGF),mammalian target of rapamycin(mTOR),threonine kinase(p-Akt),and hypoxia-inducible factor-1(HIF-1α)were observed by Western blotting.Results The expression of AQP1 and inhibition rate of cell proliferation in the baicalin group were lower than those in the control group,while the cell migration distance and apoptosis rate in the baicalin group were higher than those in the control group(all P<0.05).Cell proliferation inhibition rate was as follows:AQP1 overexpression group>control group>AQP1 down-expression group;the cell migration distance and apoptosis rate was in the following order:AQP1 down-expression group>control group>AQP1 over-expression group(all P<0.05).The expression levels of VEGF,IGF,mTOR,p-Akt,HIF-1α protein in LTEP-A2 cells were as follows:AQP1 over-expression group>AQP1 over-expression+baicalin group,AQP1 over-expression+LY294002 group,AQP1 over-expression+wortmannin group(all P<0.05),and there were no significant differences in the expression levels of various proteins between the overexpression+baicalin group,AQP1 over-expression+LY294002 group and AQP1 over-expression+wortmannin group.Conclusion Baicalin inhibits the proliferation,migration and induces the apoptosis of lung adenocarcinoma cells through down-regulating the expression of AQP1,and its mechanism may be related to the inhibition of activation of PI3K/Akt signaling pathway.
作者
韦小白
沈莹
陈静
田伟平
WEI Xiaobai;SHEN Ying;CHEN Jing;TIAN Weiping(Shanghai Jing'an District Central Hospital,Shanghai 200040,China)
出处
《山东医药》
CAS
2023年第5期6-10,共5页
Shandong Medical Journal
基金
国家自然科学基金项目(81903860)。
