黏液瘤病毒对裸鼠子宫内膜癌移植瘤的抑瘤作用及机制

目的 探讨黏液瘤病毒对裸鼠皮下移植子宫内膜癌的抑瘤作用及其可能机制.方法 采用皮下注射子宫内膜癌HEC-IB细胞的方法建立裸鼠子宫内膜癌模型.瘤内注射黏液瘤病毒后,观察肿瘤生长变化,并用免疫组化方法检测肿瘤组织中PI3K、Akt、GSK表达水平及活性的变化.同时通过瘤内注射灭活病毒和生理盐水作为对照组.结果 黏液瘤病毒对正常裸鼠不存在毒性及致病能力.黏液瘤处理组肿瘤大小较对照组缩小,差异具有统计学意义.经黏液瘤病毒处理的肿瘤组织中PI3K、Akt的表达水平较对照组显著降低.结论 黏液瘤病毒对裸鼠子宫内膜癌移植瘤具有抑瘤作用,其可能机制是通过调节PI3K/AKT（磷脂酰肌醇3激酶/丝氨酸苏氨酸激酶）信号通路调控细胞凋亡.     

天麻素保护过氧化氢诱导的神经元氧化损伤的研究

目的:探究天麻素对过氧化氢引起的神经细胞氧化损伤的保护作用及其作用机制。方法:培养SH-SY5Y细胞,并分为3组:对照组、氧化损伤组和天麻素处理组。用流式细胞术检测细胞凋亡率及细胞活性氧(ROS)水平,用ELISA检测细胞内超氧化物歧化酶(SOD)表达水平,用蛋白质免疫印迹实验检测细胞内PI3K、AKT磷酸化水平。结果:过氧化氢处理引起细胞凋亡率升高,胞内ROS平上升,SOD表达下调以及PI3K、AKT磷酸化水平下降;天麻素处理后可以显著抑制过氧化氢引起的细胞凋亡、ROS水平上升及SOD表达下调,并缓解氧化损伤对PI3K/AKT磷酸化的抑制作用。结论:天麻素通过激活PI3K/AKT信号通路保护过氧化氢诱导的神经细胞氧化损伤。     

杜仲绿原酸通过PI3K/AKT/Nrf-2通路增强视网膜母细胞瘤细胞系HXO-Rb44放射敏感性

目的　探究杜仲绿原酸（CGA）通过调控PI3K/AKT/Nrf-2通路,对视网膜母细胞瘤细胞系HXO-Rb44放射敏感性的增强作用。 方法　通过不同浓度的CGA处理HXO-Rb44细胞,检测IC10,作为后续实验中CGA浓度。将细胞分为Ctrl、Radio、CGA、Radio+CGA组,Radio组给予4 MV X射线照射24 h,CGA组使用CGA处理24 h,Radio+CGA组在CGA处理24 h后再给予4 MV X射线照射24 h,流式细胞术检测细胞周期与细胞凋亡,蛋白质印迹检测Ki67、依赖还原型辅酶I/II醌氧化还原酶I（NQO1）、活化型半胱天冬酶（cl-caspase-3）、硫氧还蛋白还原酶1（TrxR1）、活化磷脂酰肌醇3-激酶（p-PI3K）、蛋白激酶B（AKT）、p-AKT、核因子E2相关因子2（Nrf2）蛋白表达。进一步通过PI3K激活剂740Y-P处理细胞,检测细胞周期、凋亡、增殖和相关蛋白表达、PI3K/AKT/Nrf2通路蛋白表达。 结果　随着CGA浓度的增高,细胞的相对存活率降低,药物毒性呈浓度依赖性,IC10为81.59 μmol/L。与Ctrl组相比,Radio和CGA组G2/M期细胞比例显著升高,细胞凋亡比率显著升高（P<0.01）;与Radio组相比,Radio+CGA组G2/M期细胞比例和细胞凋亡比率进一步显著升高（P<0.01）。同时在蛋白水平上,与Ctrl组相比,Radio和CGA组Ki67、NQO1、TrxR1、p-PI3K、Nrf2蛋白表达及p-AKT/AKT比率显著降低（P<0.01）;放射和CGA联合作用进一步降低以上各指标（P<0.01）。此外,PI3K激活剂可逆转放射对细胞周期、增殖、凋亡以及PI3K/AKT/Nrf2通路的作用,CGA可恢复放射引起的上述各指标的变化（P<0.01）。 结论　CGA可增强HXO-Rb44细胞的放射敏感性,其作用机制与抑制PI3K/AKT/Nrf2通路相关。     

瘦素对小鼠胸腺细胞凋亡的保护机制

目的 观察瘦素对小鼠胸腺细胞凋亡的保护机制,探讨磷酸肌醇3位羟基激酶/AKT(P13-K/AKT)和胞桨型磷脂酶XZ(cPLA2)信号转导通路是否参与瘦素保护机制.方法 以IPS诱导小鼠胸腺细胞凋亡为模型,采用Annexin V-FTTC/PI双染色法流式细胞仪检测PI3-K/AKT特异性抑制剂LY294002 和 cPLA2 抑制剂AACOCF3对细胞凋亡的影响,以RT-PCR检测cPLA2 mRNA和caspase3 mRNA的表达,用cPLA2活性试剂盒检测cPLA2的活性.结果 瘦素可以保护LPS诱导的小鼠胸腺细胞凋亡,使胸腺细胞存活率由58%上升到65%,LY294002(10 μmol/L)和AACOCF3(10 μmol/L)可以明显抑制瘦素的保护作用,使胸腺细胞的存活率分别降至60%(P＜0.05)和62%(P＜0.05).RT-PCR表明加入瘦素后cPLA2 mRNA和caspase3 mRNA的表达下降.cPLA2活性试剂盒表明leptin可抑制cPLA2的活性.结论 瘦素抑制LPS诱导的胸腺细胞凋亡同时依赖IP3K/AKT和cPLA2途径.     

PI3K/AKT信号通路在全肝缺血再灌注大鼠肺损伤中的作用

目的 探讨磷脂酰肌醇-3激酶（PI3K）/AKT通路在大鼠全肝缺血再灌注肺损伤中作用。方法 本实验分两部分。(1)36只大鼠分别于肝缺血前与再灌注后不同时点处死取肺。（2）12只大鼠分成Wortmannin组与模型对照组。分别应用Western blot、原位末端转移酶(TUNEL)法与免疫组化法检测肺AKT、磷酸化AKT（p-AKT）表达、细胞凋亡与增殖细胞核抗原（PCNA）表达。结果（1）与缺血前相比,缺血再灌注后肺细胞凋亡指数显著增高; p-AKT/AKT与PCNA阳性指数呈双相变化;肺病理改变严重。（2）p-AKT/AKT与PCNA阳性指数成正相关;与凋亡指数呈负相关。（3)与模型对照组相比,Wortmannin组p-AKT/AKT与PCNA阳性指数显著降低,细胞凋亡指数显著增高,病理改变加重。结论 PI3K/AKT通路可能通过抑制凋亡、促进增殖对全肝缺血再灌注肺损伤发挥保护作用。     

PI3K/AKT途径在HGF介导肝干细胞抗凋亡调控中的作用

目的研究PI3K(磷脂酰肌醇3激酶)和MAPK(丝裂原活化蛋白激酶)途径如何参与肝干细胞的凋亡调控；探讨PI3K和MAPK信号途径在HGF介导的抗凋亡信号中的调控。方法用大鼠肝干细胞系WBF-344细胞进行实验；用流式细胞仪检测细胞及DNA凝胶电泳及流式细胞仪检测细胞凋亡；用Western blot方法检测磷酸化MAPK及PI3K蛋白的表达。结果发现TNF-α对ActD致敏的WBF-344细胞能诱导凋亡，并且这种细胞凋亡呈现剂量效应关系；HGF对ActD／TNF—α诱导Vd3F-344细胞凋亡具有保护作用，并且这种保护作用呈剂量效应关系；Western blot结果显示，在WB细胞，HGF确实能够激活ERK、p38MAPK、PI3K／AKT信号转导途径；进一步用特异的抑制剂阻断ERK及p38MAPK途径后，并不能改变HGF对TNF-α诱导凋亡的拮抗作用，而阻断PI3K／AKT途径后，HGF的抗凋亡作用被抑制。结论TNF-α能诱导ActD致敏的肝干细胞发生凋亡，而HGF则对这种细胞凋亡有明显的拮抗作用；ERK1／2和p38MAPK途径的激活并不参与HGF介导的抗凋亡作用，HGF的抗凋亡作用是通过PI3K途径转导的。     

羟氯喹通过PI3K/Akt通路对系统性红斑狼疮外周血单个核细胞凋亡的作用研究

目的：探讨羟氯喹对SLE患者外周血单个核细胞的凋亡作用及其相关机制。方法：对30例活动期SLE患者及15例正常健康人抽血分离PBMCs细胞进行培养,分为正常对照组、SLE组、羟氯喹5 mg/L、羟氯喹25 mg/L组,MTT法检测细胞生长抑制,采用Annexin V/PI流式细胞仪细胞检测凋亡率,Western blot方法检测BAX、BCL-2、PI3K、pAKt及mTOR等相关蛋白的表达影响。同时加入羟氯喹HCQ 25 mg/L和PI3K/AKT通路抑制剂LY294002 20 μmol/L,作用SLE患者的PBMCs细胞48 h,检测PBMCs细胞生长抑制和凋亡率。结果：SLE组比正常对照组PBMCs细胞生长抑制率和凋亡率显著升高（P<0.05）;与SLE组相比,羟氯喹5 mg/L和25 mg/L组细胞生长抑制率和凋亡率显著升高（P<0.05）。羟氯喹组与SLE组比较,PI3K、pAKt、mTOR、bcl-2的表达明显下降,差异有统计学意义（P<0.05）,bax和caspase-3的表达增加,差异有统计学意义（P<0.05）。PI3K/AKT抑制剂 LY294002能够阻断羟氯喹导致的SLE患者PBMCs细胞凋亡。结论：羟氯喹能够通过PI3K/Akt信号通路促进SLE患者体外PBMCs的凋亡。     

苦瓜蛋白诱导K562细胞凋亡及其对Bcl-2、PCNA蛋白表达的影响

目的：观察苦瓜蛋白是否具有诱导K562细胞凋亡的生物学活性，探讨苦瓜蛋白对K562细胞Bcl-2及PCNA表达水平的影响及其作用的分子机制。方法：以一定浓度的苦瓜蛋白处理K562细胞12～72小时，CCK-8检测其对细胞生长的影响；流式细胞术（AnnexinV）、细胞形态学（光镜及电镜）等方法检测细胞凋亡；同时应用流式细胞术检测Bcl-2及PCNA蛋白表达的变化。结果：苦瓜蛋白对K562细胞生长具有明显的抑制作用；流式细胞术及形态学观察（光镜及电镜）证实一定作用浓度的苦瓜蛋白可诱导K562细胞发生明显的细胞凋亡，且随着作用时间的延长细胞凋亡率逐渐升高。苦瓜蛋白处理组K562细胞中Bcl-2及PCNA蛋白的表达分别为0．33％和98．36％，对照组分别为74．03％和97．63％。结论：苦瓜蛋白对K562细胞生长具有明显抑制作用，其作用主要通过诱导K562细胞产生细胞凋亡．而不是抑制细胞增殖。苦瓜蛋白诱导K562细胞凋亡过程中，Bcl-2蛋白表达的下调对调控细胞凋亡有重要作用。     

PI3K/AKT通路介导H_2O_2预处理减轻PC12细胞氧化损伤

目的探讨PI3K/AKT信号通路是否参与H2O2预处理诱导的适应性细胞保护作用。方法体外培养PC12细胞,建立H2O2预处理对抗高浓度H2O2诱导细胞损伤的实验模型。应用甲氮甲唑蓝(MTT)法测定细胞的存活率,比色法测定乳酸脱氢酶(LDH)的活性,碘化丙啶(PI)染色流式细胞术检测细胞凋亡率,免疫印迹法(Westernblot)测定AKT的表达。结果 100μmol H2O2预处理PC12细胞90 min可显著地抑制300μmol H2O2引起的损伤,使细胞存活率从50.2%±4.6%升高至83.8%±3.5%,LDH活性由103%±10.2%下降至68.5%±5.3%,细胞凋亡率由65.5%±4.1%下降至37.1%±2.3%(P<0.01)。100μmol H2O2预处理诱导p-AKT的表达,PI3K抑制剂ly294002阻断了H2O2预处理引起的p-AKT表达。同时ly294002拮抗了H2O2预处理诱导的抗细胞损伤和凋亡作用。结论 H2O2预处理通过PI3K途径引起AKT的活化,PI3K/AKT通路的活化介导了H2O2预处理诱导的适应性细胞保护作用。     

自噬在红景天甙诱导胃癌细胞凋亡中的作用及机制

目的:探讨自噬在红景天甙诱导人胃癌细胞凋亡中的作用及机制。方法:红景天甙处理人胃癌AGS细胞,Hoechst33342染色观察凋亡形态学变化,流式细胞术检测凋亡细胞数;透射电镜观察自噬体形成,mRFP-GFP-LC3转染观察荧光自噬斑点;Western blot观察凋亡蛋白、自噬蛋白及PI3K/AKT/mTOR通路蛋白表达;自噬抑制剂氯喹(CQ)预处理AGS细胞,流式细胞术检测凋亡细胞数,Western blot观察凋亡相关蛋白变化,Hoechst33342染色观察凋亡形态学变化;PI3K/AKT激动剂胰岛素样生长因子(IGF-1)预处理AGS细胞,mRFP-GFP-LC3转染和Western blot分别观察自噬变化。结果:红景天甙诱导人胃癌AGS细胞自噬体形成,荧光自噬斑点增多,抑制PI3K/AKT/mTOR通路蛋白活化;CQ预处理胃癌细胞后,细胞凋亡数量显著增加,细胞核固缩现象增强,抗凋亡蛋白Bcl-2表达下调,促凋亡蛋白Bax、cleaved-Caspase3、cleaved-Caspase9表达上调;IGF-1预处理胃癌细胞明显减弱红景天甙诱导的自噬现象。结论:中药红景天甙可诱导人胃癌AGS细胞自噬,与抑制PI3K/AKT/mTOR通路相关,抑制自噬可促进红景天甙诱导的细胞凋亡。     

COX-2表达下调抑制人结肠癌细胞系HT-29增殖并促进凋亡

目的探讨环氧化酶-2(COX-2)表达对大肠癌细胞HT-29细胞的细胞活力、增殖和凋亡等生物学行为的影响。方法利用脂质体将siRNA COX-2及空载体转入HT-29大肠癌细胞中,并设立对照组。48 h后,real-time PCR法检测COX-2、Bcl-2、Bax和基质金属蛋白酶2(MMP2)mRNA表达;Western blot检测COX-2及p-AKT表达;CCK-8法检测细胞活力;Annexin V/PI流式双染检测细胞凋亡;流式细胞计量术检测细胞周期;划痕实验及Transwell实验分别检测细胞迁移和侵袭能力。结果与对照组及空载体组比较,COX-2抑制组COX-2蛋白及mRNA表达量显著降低(P0.01);细胞活力下降(P0.01);细胞凋亡率提高,细胞周期阻滞于G期,细胞迁移及侵袭能力降低(P0.01);Bcl-2及MMP2 mRNA表达下调(P0.01);Bax mRNA表达上调(P0.01);p-AKT蛋白表达下调(P0.01)。结论 COX-2表达量下调后能显著的抑制细胞增殖、迁移与侵袭,并诱导细胞凋亡,可能与下调p-AKT有关。     

血小板生成素thrombopoietin对神经保护作用的体外研究

目的体外研究血小板生成素（TPO,一种调控巨核细胞和血小板生成的因子）,对神经祖细胞C17.2的保护作用。方法建立无血清条件下C17.2细胞凋亡的模型,用TPO不同浓度处理C17.2细胞,利用MTT、Western blotting、流式细胞技术等方法检测TPO对C17.2细胞的保护作用。结果不同浓度的TPO（0、1、10、50、100、200ng/ml）都能促进C17.2细胞增殖,并且具有剂量依赖性。TPO使磷酸化AKT水平增加,促进神经祖细胞增殖,LY294002可以阻止细胞的增殖。用流式细胞仪方法检测表达Annexin V的细胞减少。结论体外研究显示TPO通过激活PI3K/AKT信号通路,对神经祖细胞C17.2起保护作用,这一发现为临床上神经损伤的治疗提供了新的思路。     

Suppressive effect of platycodin D on bladder cancer through microRNA-129-5p-mediated PABPC1/PI3K/AKT axis inactivation

Platycodin D (PD) is a major constituent of Platycodon grandiflorum and has multiple functions in disease control. This study focused on the function of PD in bladder cancer cell behaviors and the molecules involved. First, we administered PD to the bladder cancer cell lines T24 and 5637 and the human uroepithelial cell line SV-HUC-1. Cell viability and growth were evaluated using MTT, EdU, and colony formation assays, and cell apoptosis was determined using Hoechst 33342 staining and flow cytometry. The microRNAs (miRNAs) showing differential expression in cells before and after PD treatment were screened. Moreover, we altered the expression of miR-129-5p and PABPC1 to identify their functions in bladder cancer progression. We found that PD specifically inhibited the proliferation and promoted the apoptosis of bladder cancer cells; miR-129-5p was found to be partially responsible for the cancer-inhibiting properties of PD. PABPC1, a direct target of miR-129-5p, was abundantly expressed in T24 and 5637 cell lines and promoted cell proliferation and suppressed cell apoptosis. In addition, PABPC1 promoted the phosphorylation of PI3K and AKT in bladder cancer cells. Altogether, PD had a concentration-dependent suppressive effect on bladder cancer cell growth and was involved in the upregulation of miR-129-5p and the subsequent inhibition of PABPC1 and inactivation of PI3K/AKT signaling.     

敲减FGFR1基因表达对婴幼儿血管瘤内皮细胞生物学特性的影响

目的:研究敲减成纤维细胞生长因子受体1(FGFR1)基因的表达对婴幼儿血管瘤内皮细胞(HemECs)活力、凋亡、侵袭和迁移的影响。方法:经转染FGFR1小干扰RNA(si-FGFR1)下调FGFR1表达,研究FGFR1对HemECs生物学特性的影响,以CCK-8法检测细胞活力的变化,流式细胞术检测细胞的凋亡水平, Transwell实验检测细胞侵袭和迁移能力的改变;Western blot法检测磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)和磷酸化AKT(p-AKT)的蛋白水平。结果:转染si-FGFR1进入HemECs能够显著抑制细胞的活力(P0.05),促进其凋亡(P0.05),降低细胞的侵袭和迁移能力(P0.05);Western blot结果显示,下调细胞中FGFR1基因的表达可降低PI3K和p-AKT的蛋白水平(P0.05),但对AKT蛋白水平无显著影响。结论:敲减HemECs细胞内FGFR1的表达可通过影响PI3K/AKT信号通路调节婴幼儿血管瘤内皮细胞的生物学特性。     

FTY720 attenuates hypoxia–reoxygenation-induced apoptosis in cardiomyocytes

FTY720, sphingosine 1 phosphate (S1P) receptor agonist, is a potent immunosuppressive agent. Numerous studies have documented a relationship between S1P and cardioprotection. We therefore hypothesized that a S1P analogue FTY720 would attenuate hypoxia/reoxygenation (H/R) induced cadiomyocyte apoptosis. H9C2 cardiomyocytes were employed to establish an in vitro model of H/R. Cells were treated or not with different doses of FTY720. Cell viability was measured by flow cytometry and TUNEL staining. Western blot was used to analyze downstream signaling pathway. We observed that FTY720 inhibits the expression of cleaved caspase-3 and activates both AKT and ERK1/2 pathways. AKT pathway can be blocked by MEK kinase inhibitor PD98059. ERK1/2 pathway can be blocked by the phosphoinositide-3 kinase inhibitor wortmannin. AKT and ERK1/2 activation can also be inhibited by S1P1/3 receptor antagonist VPC23019, Gi antagonist PTX. The protein levels of TNF-α and IL1ß were upregulated during hypoxia/reoxygenation and were attenuated by FTY720. We conclude that FTY720, via its cargo of S1P, can protect cardiomyocytes against hypoxia/reoxygenation injury. This effect is achieved by inhibiting caspase-3 expression, inflammatory cytokine levels and activating AKT and ERK1/2 signaling pathways. The prosurvival signal activation is dependent on S1P1, 3 subtype receptors and Gi protein.     

赖氨大黄酸通过激活JNK诱导宫颈癌HeLa细胞凋亡

目的研究赖氨大黄酸(RHL)对人宫颈癌HeLa细胞增殖、凋亡的影响,并探讨其作用机制。方法用MTT法检测细胞增殖;用流式细胞仪检测细胞凋亡,用Western blot检测凋亡相关蛋白及JNK蛋白与蛋白磷酸化水平。结果 RHL能有效抑制宫颈癌HeLa细胞增殖,并能诱导其凋亡,随药物浓度的增加,细胞凋亡率也逐渐升高;RHL能够激活caspase-3、caspase-7和PARP,并激活磷酸化的JNK表达,磷酸化的JNK表达增加在RHL的诱导凋亡中起主要作用。结论 RHL通过激活JNK-caspase-PARP信号通路抑制HeLa细胞增殖,并诱导其凋亡,赖氨大黄酸解决了大黄酸不溶于水的问题,有望成为临床肿瘤辅助化疗药物。     

Activation of the phosphatidylinositol 3'-kinase/AKT pathway in neuroblastoma and its regulation by thioredoxin 1

Neuroblastoma is a malignant pediatric tumor with poor survival. The phosphatidylinositol 3'-kinase/AKT pathway is a crucial regulator of cellular processes including apoptosis. Thioredoxin 1, an inhibitor of tumor-suppressor phosphatase and tensin homolog, is overexpressed in many tumors. The objective of this study was to explore phosphatidylinositol 3'-kinase/AKT pathway activation and regulation by thioredoxin 1 to identify potential therapeutic targets. Immunohistochemical analysis was done on tissue microarrays from tumor samples of 101 patients, using antibodies against phosphatidylinositol 3'-kinase, AKT, activated AKT, phosphatase and tensin homolog, phosphorylated phosphatase and tensin homolog, thioredoxin 1, epidermal growth factor receptor, vascular endothelial growth factor and receptors (vascular endothelial growth factor 1 and vascular endothelial growth receptor 2), platelet-derived growth factor receptors, insulin-like growth factor 1 receptor, neurotrophic tyrosine kinase receptor type 2, phosphorylated 70-kd S6 protein kinase, 4E-binding protein 1, and phosphorylated mammalian target of rapamycin. Using 3 neuroblastoma cell lines, we investigated cell viability with AKT-specific inhibitors (LY294002, RAD001) and thioredoxin 1 alone or in combination. We found activated AKT and AKT expressed in 97% and 98%, respectively, of neuroblastomas, despite a high expression of phosphatase and tensin homolog correlated with thioredoxin 1. AKT expression was greater in metastatic than primary tumors. Insulin-like growth factor 1 receptor, tyrosine kinase receptor type 2, vascular endothelial growth receptor 1, and downstream phosphorylated 70-kd S6 protein kinase were correlated with activated AKT. LY294002 and RAD001 significantly reduced AKT activity and cell viability and induced a G(1) cell cycle arrest. Thioredoxin 1 decreased cytotoxicity of AKT inhibitors and doxorubicin, up-regulated AKT activation, and induced cell growth. Thus, vascular endothelial growth receptor 1, tyrosine kinase receptor type 2, insulin-like growth factor 1 receptor, and thioredoxin 1 emerged as preferentially committed to phosphatidylinositol 3'-kinase/AKT pathway activation as observed in neuroblastoma. Thioredoxin 1 is a potential target for therapeutic intervention.     

Modulatory role of garlicin in migration and invasion of intrahepatic cholangiocarcinoma via PI3K/AKT pathway

Increasing evidences have indicated the role of garlicin in inhibiting the progression of various tumors including glioma, pulmonary carcinoma and pancreatic carcinoma, via mediating cell apoptosis or cell cycle. The regulatory effect and related molecular mechanism of garlicin in intrahepatic cholangiocarcinoma, however, remained unknown. This study thus aimed to investigate this scientific issue. HCCC-9810 cell line was treated with serially diluted garlicin, followed by cell proliferation assay using MTT approach. Transwell migration and invasion assays were further employed the regulatory effect of garlicin. The expression level of p-AKT and AKT proteins in tumor cells was quantified by Western blot. The growth of tumor cells was significantly inhibited by high concentration of garlicin (> 1.5 μM). Lower concentration of garlicin showed dose-dependent inhibition of tumor cell invasion and migration. After using specific agonist IGF-1 (50 ng/mL) of PI3K/AKT signaling pathway, such facilitating effects of garlicin were depressed (P < 0.05). Western blotting showed significantly decreased phosphorylation level of AKT after treated with gradient concentrations of garlicin, while leaving the total AKT protein level unchanged. Garlicin may inhibit the invasion and migration of intrahepatic cholangiocarcinoma cells via inhibiting PI3K/AKT signaling pathway.