索拉非尼联合麦冬皂苷D`共同抑制ERK和AKT通路促进PC3细胞凋亡  

Sorafenib combined with ophiopogonin D` inhibits ERK and AKT pathways to promote apoptosis in androgen-independent prostate cancer PC3 cells

作　　者：张耀文 卢宗亮[1] 李龙 许红霞[1] ZHANG Yaowen;LU Zongliang;LI Long;XU Hongxia(Department of Clinical Nutrition,Daping Hospital,Army Medical University(Third Military Medical University),Chongqing,400042,China)

机构地区：[1]陆军军医大学(第三军医大学)大坪医院临床营养科,重庆400042

出　　处：《第三军医大学学报》2021年第19期1870-1876,共7页Journal of Third Military Medical University

基　　金：重庆市自然科学基金面上项目(cstc2020jcyj-msxmX0499)。

摘　　要：目的探讨索拉非尼(sorafenib, Sor)联合麦冬皂苷D`(Ophiopogonin D`,OPD`)对雄激素非依赖前列腺癌细胞(PC3细胞)的抗肿瘤效应及其机制。方法在体外实验中,Western blot检测不同浓度的OPD`(0、1.0、2.5、5.0μmol/L)、Sor(0、2.5、5.0、10.0μmol/L)、OPD`(2.5μmol/L)+Sor(5.0μmol/L)联合使用24 h后,对PC3细胞AKT、p-AKT、ERK、p-ERK表达的影响;流式细胞仪Annexin V-FITC和PI双染检测细胞凋亡。在体内实验中,构建PC3细胞异种移植瘤模型后,将裸鼠分为对照组、OPD`组(1.0 mg/kg)、Sor组(2.5 mg/kg)、OPD`+Sor联合使用组并进行相应干预,且每3天测量裸鼠体质量和肿瘤体积,至药物干预21 d。结果 Sor主要下调p-AKT蛋白表达及p-AKT/AKT水平(P<0.05),仅在10.0μmol/L时,降低p-ERK/ERK水平(P<0.05)。OPD`主要下调p-ERK蛋白的表达及p-ERK/ERK水平(P<0.05),仅在5.0μmol/L时,降低p-AKT/AKT水平(P<0.05)。2.5μmol/L OPD`和5.0μmol/L Sor联用时,p-AKT和p-ERK的蛋白表达及p-AKT/AKT和p-ERK/ERK水平均显著降低(P<0.05)。细胞凋亡检测结果显示:两药联用组的凋亡指数明显高于OPD`或Sor单独用药组(P<0.05)。体内实验结果显示:1.0 mg/kg OPD`和2.5 mg/kg Sor联用组肿瘤体积和质量均低于其他组(P<0.05)。结论索拉非尼联合麦冬皂苷D`通过抑制ERK和AKT通路,促进雄激素非依赖前列腺癌细胞凋亡。Objective To investigate the anti-tumor effect of sorafenib(Sor) combined with Ophiopogonin D`(OPD`) on androgen-independent prostate cancer(PC3) cells and its mechanism. Methods In vitro, the expression of AKT, p-AKT, ERK and p-ERK in PC3 cells was detected by Western blotting in 24 h after being treated with different concentrations of OPD`(0, 1, 2.5 and 5.0 μmol/L), Sor(0, 2.5, 5.0 and 10.0 μmol/L) and OPD`(2.5 μmol/L)+Sor(5.0 μmol/L), respectively. Apoptosis of PC3 cells was observed by flow cytometry with AnnexinV-FITC and propidium iodide(PI) double staining. In vivo, PC3 cells xenograft tumor model was constructed in nude mice. Subsequently, 24 nude mice were equally divided into control group, OPD` group(1.0 mg/kg), Sor group(2.5 mg/kg) and OPD` +Sor combined group. The body weight of nude mice and the tumor volume were measured and recorded every 3 days until 21 d after drug intervention. Results Sor mainly down-regulated the expression levels of p-AKT and p-AKT/AKT(P<0.05), and only decreased p-ERK/ERK level at the concentration of 10.0 μmol/L(P<0.05). While OPD` primarily down-regulated p-ERK and p-ERK/ERK levels(P<0.05), and only reduced p-AKT/AKT at 5.0 μmol/L(P<0.05). However, the protein levels of p-AKT, p-ERK, p-AKT/AKT, and p-ERK/ERK were all greatly diminished in the combined group(2.5 μmol/L OPD` +5.0 μmol/L Sor)(P<0.05). In addition, the apoptotic index was significantly higher in the combined group than the OPD` or Sor alone group(P<0.05). The results of in vivo experiments also showed that the tumor volume and tumor weight in the combined group(1.0 mg/kg OPD` +2.5 mg/kg Sor) were much lower than those in the other groups(P<0.05). Conclusion Sor combined with OPD` promotes apoptosis in PC3 cells by inhibiting ERK and Akt pathways.

关 键 词：索拉非尼 麦冬皂苷D` 雄激素非依赖前列腺癌 ERK AKT 细胞凋亡 

分 类 号：R737.25[医药卫生—肿瘤] R966[医药卫生—临床医学]