半枝莲含药血清通过抑制mTOR信号通路影响非小细胞肺癌NCI-H1650细胞上皮间质转化

Effect of Banzhilian(Scutellariae Barbatae Herba)Drug-Containing Serum on Epithelial-Mesenchymal Transition of NSCLC NCI-H1650 Cells by Inhibiting mTOR Signaling Pathway

目的探讨半枝莲含药血清通过抑制雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路影响非小细胞肺癌(non-small cell lung cancer,NSCLC)NCI-H1650细胞上皮间质转化。方法选取健康青紫蓝大白兔制备半枝莲含药血清,根据不同剂量分为对照组、低剂量组(A组)、中剂量组(B组)、高剂量组(C组)。选取人非小细胞肺癌细胞NCI-H1650与含药血清培养,采用逆转录RT-PCR、免疫印迹、MTT、划痕实验等方法,观察4组不同剂量半枝莲含药血清抑制mTOR信号通路对NCI-H1650细胞上皮间质转化、增殖、迁移的影响。结果4组mTOR表达量差异具有统计学意义,其中C组mTOR表达量低于对照组和A组(P<0.05);4组Vimentin、N-cadherin、E-cadherin蛋白表达相对灰度值差异具有统计学意义,且C组Vimentin、N-cadherin灰度值低于对照组、A组、B组,E-cadherin灰度值高于对照组(P<0.05);4组24、48、72 h吸光度差异具有统计学意义,且C组24、48、72 h吸光度低于对照组、A组、B组(P<0.05);4组NCI-H1650细胞迁移率差异具有统计学意义,且A组、B组、C组迁移率低于对照组,C组低于A组、B组(P<0.05)。结论高剂量半枝莲含药血清可通过抑制mTOR信号通路影响NCI-H1650细胞上皮间质转化、增殖及迁移,从而抑制NSCLC发生和发展。

Objective To investigate the effect of Banzhilian(Scutellariae Barbatae Herba)drug-containing serum on epithe⁃lial-mesenchymal transition(EMT)of non-small cell lung cancer(NSCLC)NCI-H1650 cells by inhibiting rapamycin target protein(mTOR)signaling pathway.Methods Healthy white rabbits were divided into a control group,a low dose group(group A),a medium dose group(group B)and a high dose group(group C).Human non-small cell lung cancer cell line NCI-H1650 was cultured with medicated serum.The effects of inhibiting mTOR signaling pathway by four groups of different doses of Banzhilian(Scutellariae Barbatae Herba)drug-containing serum on epithelial-mesenchymal transition,proliferation and migra⁃tion of NCI-H1650 cells were observed by reverse transcription-RT-PCR,immunoblotting,MTT and scratch assay.Result⁃s There were significant differences in mTOR expression among the four groups.The mTOR expression in group C was lower than that in the control group and the group A(P<0.05).There were significant differences in the relative gray values of vimentin,N-cadherin and E-cadherin protein expression among the four groups,and the gray values of vimentin and N-cadherin in group C were lower than those in the control group,the group A and the group B,while the gray values of E-cadherin were high⁃er than those in the control group(P<0.05).There were significant differences in the absorbance at 24 h,48 h and 72 h among the four groups,and the absorbance at 24 h,48 h and 72 h in group C was lower than that in the control group,the group A and the group B(P<0.05).There were significant differences in the migration rates of NCI-H1650 cells among the four groups,and the migration rates in group A,B and C were lower than those in the control group.Conclusions High dose of Banzhilian(Scutellariae Barbatae Herba)drug-containing serum can inhibit the occurrence and development of NSCLC by inhibiting mTOR signaling pathway and affecting epithelial-mesenchymal transition,proliferation and migration of NCI-H1650 cells.