精氨酸脱亚胺酶阻断PI3K-AKT通路抑制胰腺癌细胞侵袭  

Arginine deiminase inhibits pancreatic cancer cell invasion by blocking PI3-KAKT signaling pathway

作　　者：刘江波[1] 雷亮亮[1] 杨延辉[1] 黎韡[2] 马清涌[2] 刘德纯[1] 李三强[3] 

机构地区：[1]河南科技大学临床医学院河南科技大学第一附属医院普通外科,河南省洛阳市471000 [2]西安交通大学医学院第一附属医院肝胆外科,陕西省西安市710061 [3]河南科技大学医学院肝脏损伤与修复分子医学重点实验室,河南省洛阳市471003

出　　处：《世界华人消化杂志》2016年第24期3570-3579,共10页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;Nos.U1504815;81301846~~

摘　　要：目的研究精氨酸脱亚胺酶(arginine deiminase,ADI)对胰腺癌细胞株迁移、侵袭能力的影响及可能的分子机制.方法选取精氨琥珀酸合成酶表达缺陷和表达阳性的胰腺癌细胞株PANC-1和BxPC-3接受含ADI培养基或普通培养基培养干预.细胞划痕及Transwell试验检测细胞迁移和侵袭能力;实时定量PCR技术及Western blot检测侵袭相关基因mRNA和蛋白的改变.Western blot和Transwell试验检测ADI联合PI3K信号抑制剂LY29400对PANC-1细胞侵袭行为及分子的影响.结果ADI可抑制PANC-1细胞的迁移、侵袭(P<0.05),下调PANC-1细胞尿激酶型纤溶酶原激活因子、基质蛋白金属酶(matrix metalloproteinases,MMP)-2、MMP-9,和上调金属蛋白酶类组织抑制剂-2和E-Cadherin的m RNA和/或蛋白表达水平(P<0.05);而对BxPC-3细胞侵袭能力影响不明显.ADI可下调PANC-1细胞PI3K/AKT/核转录因子-kB(nuclear factor-kappa B,NF-kB)信号通路蛋白p-AKT、p-p65的表达水平,而LY294002则协同ADI的这种作用,并协同下调MMP-2水平;Transwell侵袭试验也显示LY294002可协同A D I抑制胰腺癌细胞的侵袭能力(P<0.05).结论ADI通过阻断PI3K-AKT信号通路调控侵袭相关基因表达抑制胰腺癌细胞侵袭.AIM To investigate the impact of arginine deiminase（ADI） on the migration and invasion of human pancreatic cancer cells and the possible mechanism involved.METHODS The pancreatic cancer cell lines PANC-1expressing defective argininosuccinate synthase（ASS） and BxPC-3 expressing ASS protein were chosen for the ADI treatment experiments,and they were cultured in the medium containing ADI（experimental group） or the common medium without ADI（control group）.The impact of ADI on the migration and invasion of the two pancreatic cancer cell lines was examined by scratch assay and transwell invasion assay.The mRNA and protein expression of invasionrelated genes in pancreatic cancer cells treated with ADI was detected by real-time quantitative PCR and/or Western blot.The expression of signal transduction proteins and invasionrelated proteins in PANC-1 cell treated with ADI in combination with PI3 K signaling inhibitor LY294002 was also analyzed.RESULTS ADI significantly inhibited cell migration and invasion（P 〈 0.05）,down-regulated the mRNA and protein levels of urokinase plasminogen activator,matrix metalloproteinases（MMP）-2,as well as MMP-9,and elevated the levels of tissue inhibitor of metalloproteinase-2 and E-Cadherin（P 〈 0.05） in ASS deficient pancreatic cancer cell line PANC-1;while there were no obvious changes for ASS-positive pancreatic cancer cell line BxPC-3.ADI reduced the expression levels of p-AKT and p-p65,which are involved in the PBK/AKT/nuclear factor-kappa B（NF-kB）signaling,in PANC-1 cells,and PBK inhibitor LY294002 can synergize the effect of ADI on reducing the levels of phosphorylation of the signaling protein and MMP-2.Furthermore,in combination with ADI,LY294002 synergistically inhibited the invasion ability of pancreatic cancer PANC-1 cells（P 〈 0.05）.CONCLUSION ADI inhibits the invasion of pancreatic cancer cells by regulating the expression of invasionrelated genes via blocking the PBK-AKT signaling pathway.

关 键 词：胰腺肿瘤 精氨琥珀酸合成酶 精氨酸剥夺 侵袭转移 PI3K信号 

分 类 号：R735.9[医药卫生—肿瘤]