苦豆碱通过抑制PI3K/AKT/mTOR信号通路调控前列腺癌细胞自噬与凋亡  

Study on the Aloperine Regulating Autophagy and Apoptosis of Prostate Cancer Cells by Inhibiting PI3K/AKT/mTOR Signaling Pathway

作　　者：安康杰 李旭 杨尉鑫 聂黎虹[3] 李亚杰 赵瑞宁[1] AN Kang-jie;LI Xu;YANG Yu-xin;NIE Li-hong;LI Ya-jie;ZHAO Rui-ning(Urology,General Hospital of Ningxia Medical University,Yinchuan 750003,China;Ningxia Gem Flower Hospital,Yinchuan 750005,China;School of Basic Medicine Sciences,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院泌尿外科,宁夏银川750003 [2]宁夏宝石花医院,宁夏银川750005 [3]宁夏医科大学基础医学院,宁夏银川750004

出　　处：《中药材》2023年第8期2016-2022,共7页Journal of Chinese Medicinal Materials

基　　金：宁夏自然科学基金资助项目(2022AAC03506,2022AAC03160)。

摘　　要：目的:探讨苦豆碱对前列腺癌PC3、LNCaP细胞的抑制作用及其可能机制。方法:采用不同浓度(0、25、50、100、200、400、800、1 600μmol)苦豆碱处理前列腺癌PC3、LNCaP细胞。CCK-8法及克隆形成实验检测PC3、LNCaP细胞增殖活性;光学显微镜观察细胞形态改变;流式细胞术检测细胞凋亡率;免疫荧光检测自噬指标LC3Ⅱ表达;Western Blot检测凋亡和自噬相关蛋白Caspase-3、Bax、Bcl-2、LC3Ⅱ、Beclin 1、P62及PI3K/AKT/mTOR通路关键蛋白表达。结果:CCK-8及克隆形成实验结果提示苦豆碱以浓度依赖性抑制PC3、LNCaP细胞活力,与空白对照组(0.1%DMSO)比较,100、200μmol苦豆碱处理后的PC3、LNCaP细胞形态发生明显变化,流式细胞术结果显示细胞凋亡率升高,免疫荧光染色结果显示PC3和LNCaP细胞胞质LC3Ⅱ绿色荧光点状表达数量增多,自噬加强,Western Blot结果显示凋亡相关蛋白Caspase-3、Bax表达显著升高,Bcl-2表达显著降低;自噬相关蛋白Beclin 1表达及LC3Ⅱ/LC3Ⅰ比值显著升高,P62表达显著降低,PI3K通路相关蛋白p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR比例降低。结论:苦豆碱通过PI3K/AKT/mTOR通路诱导前列腺癌PC3、LNCaP细胞发生自噬和凋亡,抑制细胞增殖。Objective:To investigate the inhibitory effect and its possible mechanism of aloperine on PC3 and LNCaP cells of prostatic cancer.Methods:Different concentrations of aloperine(0,25,50,100,200,400,800,1600μmol)were used to treat PC3 and LNCaP cells of prostate cancer.The proliferative activities of PC3 and LNCaP cells were detected by CCK-8 assay and clonal formation assay.Morphological changes of cells were observed by optical microscope.The apoptosis rate was detected by flow cytometry.The expression of autophagy index LC3Ⅱwas detected by immunofluorescence.The expressions of apoptosis and autophagy related proteins Caspase-3,Bax,Bcl-2,LC3Ⅱ,Beclin 1,P62 and key proteins in PI3K/AKT/mTOR pathway were detected by Western Blot.Results:The results of CCK-8 and clonal formation experiments showed that aloperine inhibited the activities of PC3 and LNCaP cells in a concentration-dependent manner.Compared with the normal control group(0.1%DMSO),the cell morphology of PC3 and LNCaP cells treated with 100 and 200μmol aloperine was significantly changed.Flow cytometry results showed that the apoptosis rate was increased.Western Blot results showed that the expressions of apoptosis-related proteins Caspase-3 and Bax were significantly increased,and the expression of Bcl-2 was significantly decreased.The expressions of autophagy related protein Beclin 1 and the ratio of LC3Ⅱ/LC3Ⅰwere significantly increased,while the expression of P62 was decreased.The ratio of PI3K pathway related proteins p-PI3K/PI3K,pAKT/AKT and p-mTOR/mTOR decreased.Conclusion:Aloperine induces autophagy and apoptosis of PC3 and LNCaP cells of prostate cancer through PI3K/AKT/mTOR pathway and inhibits cell proliferation.

关 键 词：苦豆碱 前列腺癌 PI3K AKT MTOR 自噬 

分 类 号：R285.5[医药卫生—中药学]