补肺汤靶向调控PI3K/AKT/mTOR信号通路抑制骨肉瘤细胞增殖及侵袭机制

Mechanism of Bufei Decoction Targeted Regulating PI3K/AKT/mTOR Signaling Pathway to Inhibit the Proliferation and Invasion of Osteosarcoma Cells

[目的]研究补肺汤对骨肉瘤细胞增殖及侵袭的影响,探讨补肺汤对骨肉瘤的防治作用及其潜在机制。[方法]将大鼠骨肉瘤细胞UMR-106分为对照组,补肺汤低、中、高浓度组,对照组加入洛斯维尔帕克纪念研究所(Roswell Park Memorial Institute,RPMI)1640培养液进行培养,补肺汤低、中、高浓度组分别加入不同浓度的补肺汤进行培养。采用噻唑蓝(methyl thiazolyl tetrazolium,MTT)比色法检测细胞增殖率,划痕实验、Transwell实验探究细胞迁移能力,免疫印迹、实时荧光定量聚合酶链式反应(Real-time quantitative polymerase chain reaction,Real-time qPCR)检测磷脂酰肌醇-3-激酶(phosphatidylinositol-3-kinase,PI3K)、蛋白激酶B(protein kinase B,AKT)、哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)的蛋白及mRNA表达。[结果]与对照组比较,补肺汤各浓度组细胞增殖率明显下降(P<0.001,P<0.0001);中、高浓度组细胞迁移数量明显减少(P<0.0001);中、高浓度组穿过小室细胞数量明显减少(P<0.0001);中、高浓度组AKT、mTOR蛋白和mRNA相对表达量均明显下调(P<0.05,P<0.01,P<0.001,P<0.0001)。[结论]补肺汤具有较好的抗骨肉瘤细胞增殖迁移能力,其可能通过干预PI3K/AKT/mTOR信号通路发挥抗肿瘤作用。

[Objective] To explore the effect of Bufei Decoction on the proliferation and invasion of osteosarcoma cells and clarify the preventive and therapeutic effect and potential mechanism of Bufei Decoction on osteosarcoma. [Methods] Rat osteosarcoma cells UMR-106 were divided into control group, Bufei Decoction low, medium and high concentration groups. Control group was added with Roswell Park Memorial Institute(RPMI) 1640 culture medium for cell culture, and Bufei Decoction low, medium and high concentration groups were added with different concentrations of Bufei Decoction for culture. Methyl thiazolyl tetrazolium(MTT) assay was used to detect the cell proliferation rate, scratch test and Transwell test were used to explore the invasiveness of cells, Western blot and Real-time quantitative polymerase chain reaction(Real-time q PCR) were used to detect the expression of phosphatidylinositol-3-kinase(PI3K), protein kinase B(AKT), mammalian target of rapamycin(mTOR) protein and mRNA. [Results] Compared with control group, the cell proliferation rate of each Bufei Decoction concentration group decreased significantly(P <0.001, P <0.0001);the number of cells migrating to the scratch decreased significantly in medium and high concentration groups(P<0.0001);the number of cells passing through the chamber decreased significantly in medium and high concentration groups(P <0.0001);the relative expressions of AKT, mTOR protein and mRNA were significantly decreased in medium and high concentration groups(P<0.05, P<0.01, P<0.001, P<0.0001). [Conclusion] Bufei Decoction has a good ability to resist the proliferation, invasion and migration of osteosarcoma cells, which may play an anti-tumor role by interfering with PI3K/AKT/mTOR signaling pathway.