Cao J Y, Yin H S, Li H S, Yu X Q, Han X
Department of Urology, Binzhou Medical University Hospital, Binzhou, China.
Department of Gastrointestinal Surgery, Binzhou Medical University Hospital, Binzhou, China.
Braz J Med Biol Res. 2017 Jul 20;50(8):e6207. doi: 10.1590/1414-431X20176207.
Both sorafenib and interleukin-27 (IL-27) are antineoplastic drugs. This study aimed to investigate the synergistic effect of these two drugs on bladder cancer cells. HTB-9 and T24 cells were stimulated with IL-27 (50 ng/mL), sorafenib (2 μM) or the synergistic action of these two drugs. The cells without treatment acted as control. Cell proliferation, apoptosis and invasion were measured by bromodeoxyuridine assay, flow cytometry and modified Boyden chamber, respectively. Simultaneously, both modified Boyden chamber and scratch assay were used to assess cell migration. Finally, the phosphorylation levels of key kinases in the Akt/mechanistic target of rapamycin (mTOR)/mitogen-activated protein kinase (MAPK) pathway, and expression levels of matrix metalloproteinase (MMP)-2 and MMP-9 were detected by western blot analysis. Stimulation with IL-27 or sorafenib repressed proliferation, migration and invasion but promoted apoptosis, and the effects were all enhanced by the combination of these two drugs in HTB-9 cells. The effect of the combined treatment on bladder cancer cells was verified in T24 cells. Additionally, the phosphorylation levels of AKT, mTOR and MAPK as well as the expression levels of MMP-2 and MMP-9 were all decreased by a single treatment of IL-27 or sorafenib, and further decreased by the combined treatment of these two drugs. The combination of IL-27 and sorafenib inhibited proliferation, migration and invasion and promoted apoptosis of bladder cancer cells compared with mono-drug treatment. Additionally, the AKT/mTOR/MAPK pathway might be implicated in the functional effects by down-regulations of MMP-2 and MMP-9.
索拉非尼和白细胞介素-27(IL-27)均为抗肿瘤药物。本研究旨在探讨这两种药物对膀胱癌细胞的协同作用。用IL-27(50 ng/mL)、索拉非尼(2 μM)或这两种药物的协同作用刺激HTB-9和T24细胞。未经处理的细胞作为对照。分别通过溴脱氧尿苷测定法、流式细胞术和改良的Boyden小室检测细胞增殖、凋亡和侵袭。同时,使用改良的Boyden小室和划痕试验评估细胞迁移。最后,通过蛋白质印迹分析检测Akt/雷帕霉素机制性靶标(mTOR)/丝裂原活化蛋白激酶(MAPK)通路中关键激酶的磷酸化水平以及基质金属蛋白酶(MMP)-2和MMP-9的表达水平。用IL-27或索拉非尼刺激可抑制HTB-9细胞的增殖、迁移和侵袭,但促进其凋亡,两种药物联合使用可增强这些作用。联合治疗对膀胱癌细胞的作用在T24细胞中得到验证。此外,单独用IL-27或索拉非尼处理均可降低AKT、mTOR和MAPK的磷酸化水平以及MMP-2和MMP-9的表达水平,两种药物联合处理可进一步降低。与单药治疗相比,IL-27和索拉非尼联合使用可抑制膀胱癌细胞的增殖、迁移和侵袭并促进其凋亡。此外,AKT/mTOR/MAPK通路可能通过下调MMP-2和MMP-9参与其功能作用。