马钱苷元对人胰腺癌细胞BXPC3的抗肿瘤作用及机制研究

Study on Anti-tumor Effect of Loganetin on Pancreatic Cancer Cell BXPC3 and Its Mechanism

关键词:  山茱萸  马钱苷元  胰腺癌  侵袭和迁移  AKT-mTOR信号通路

Key words:  Corni Fructus  loganetin  pancreatic cancer  migration and invasion  AKT-mTOR signaling pathway

摘要:

目的 研究山茱萸提取物马钱苷元对人胰腺癌细胞株BXPC3的体外抗肿瘤作用，并探讨其可能的抗肿瘤机制。方法 共设置2组实验，即对照组和马钱苷元组。采用CCK-8试验检测细胞存活率的改变，流式细胞仪检测马钱苷元对细胞周期的影响。同时利用划痕试验及基质胶Transwell小室试验分别检测马钱苷元对人胰腺癌细胞株BXPC3迁移和侵袭的影响。最后通过Western blotting检测AKT-mTOR信号通路相关蛋白AKT、mTOR和S6磷酸化水平的改变。结果 与对照组相比，马钱苷元对人胰腺癌细胞株BXPC3的增殖具有抑制作用，且呈明显的时间和浓度依赖性，100 μmol·L^-1马钱苷元作用于BXPC3细胞24 h，48 h的抑制率分别为（21.49±0.01）%，（29.25±0.03）%；马钱苷元能够使细胞周期阻滞于S期并抑制细胞株BXPC3的迁移（P<0.01）和侵袭（P<0.01），下调AKT、mTOR和S6的磷酸化水平。结论 马钱苷元可能通过抑制AKT-mTOR信号通路发挥其抗肿瘤活性。

Abstract:

OBJECTIVE To investigate the anti-tumor effect of loganetin which is extract from Corni Fructus on pancreatic cancer cell BXPC3 in vitro, and its potential anti-tumor mechanism. METHODS Two groups of experiments were set up, namely the control group and the loganetin group. CCK-8 assay was performed to evaluate change of cell survival and flow cytometry was employed to analyze the effect of loganetin on cell cycle distribution. The migration of human pancreatic cancer cell line BXPC3 was examined by wound healing assay and the invasion was determined using Matrigel-transwell chamber test. Finally, the phosphorylation level of AKT, mTOR and S6 proteins in the AKT-mTOR signaling pathway were detected by Western blotting. RESULTS Compared with the control group, loganetin inhibited the proliferation of human pancreatic cancer cell line BXPC3 in a time and concentration-dependent manner, the inhibition rate of BXPC3 by 100 μmol×L^-1 loganetin for 24 h and 48 h were (21.49±0.01)% and (29.25±0.03)%. Loganetin could block cell cyle in S phase, inhibit migration(P<0.01) and invasion(P<0.01) of cell line BXPC3, and down-regulate the phosphorylation level of AKT, mTOR and S6. CONCLUSION Loganetin may exert its anticancer activity via inhibiting the AKT-mTOR signaling pathway.