SATB1通过PI3K/Akt信号通路在哌立福新对胃癌失巢凋亡抵抗细胞作用机制中的研究  

Study of SATB1 via PI3K/Akt signaling pathway in mechanism of perifosine on anoikis resistant gastric cancer cells

作　　者：秦玉萍 高迎新 马熙萌 杜瑶 王新 禹莉(指导) QIN Yu-Ping;GAO Ying-Xin;MA Xi-Meng;DU Yao;WANG Xin;YU Li(Laboratory of Oncology Basic Research and Clinical Laboratory Diagnosis,Bengbu Medical College,Bengbu 233030,China)

机构地区：[1]蚌埠医学院肿瘤基础研究与临床检验诊断重点实验室,蚌埠233030 [2]安徽省感染与免疫重点实验室,蚌埠233030

出　　处：《中国免疫学杂志》2021年第3期273-281,共9页Chinese Journal of Immunology

基　　金：安徽省自然科学基金(No.1808085MH305);国家大学生创新创业培训计划(No.201810367029);省级大学生创新创业培训计划(S202010367032)资助。

摘　　要：目的:探讨人胃癌细胞失巢凋亡抵抗的机制,研究哌立福新对胃癌细胞凋亡抵抗的可能治疗作用以及SATB1通过PI3K/Akt信号通路在哌立福新对胃癌失巢凋亡抵抗细胞中的作用机制。方法:在超低黏附条件下培养胃癌AGS亲本细胞,建立AGS细胞失巢凋亡抵抗模型,检测细胞增殖、凋亡、侵袭、迁移能力、PI3K/Akt信号通路相关蛋白以及MMP-2、SATB1的表达与亲本细胞的差异;通过MTT、流式细胞术、划痕试验、Transwell试验和Western blot等检测哌立福新对凋亡抵抗AGS细胞生物学行为的影响,抑制信号传导通路及干扰SATB1基因的表达对胃癌失巢凋亡抵抗AGS细胞增殖、凋亡、侵袭和迁移的影响。结果:成功构建了胃癌AGS细胞失巢凋亡抵抗模型。与亲代细胞相比,胃癌AGS细胞失巢凋亡模型的生长速度更快,凋亡率更低,迁移和侵袭能力更强,PIK3CA、Akt mRNA表达增加,PTEN mRNA表达减少,p-Akt、SATB1、MMP-2的蛋白表达增加,PTEN蛋白表达减少。不同浓度的哌立福新作用于凋亡抵抗AGS细胞后,抑制细胞生长,诱导细胞凋亡,抑制细胞迁移和侵袭,p-Akt、MMP-2蛋白表达随哌立福新浓度增加而下降,PTEN蛋白表达逐渐增加。此外,发现哌立福新降低了SATB1蛋白在凋亡抵抗AGS细胞中的表达,SATB1的下调增强了哌立福新介导的抗肿瘤能力。结论:哌立福新可通过PI3K/Akt途径对凋亡抵抗AGS细胞发挥抗癌作用,并且下调SATB1的表达可增强抗癌作用。本次研究可能为胃癌转移提供一种新的潜在的抗肿瘤治疗策略。Objective:To investigate the mechanism of anoikis resistance in human gastric cancer cells and study the possible therapeutic effect of perifosine and the role of SATB1 via PI3K/Akt signaling pathway in the apoptosis resistance of gastric cancer cells.Methods:AGS parental cells were cultured under ultra-low adhesion condition to establish anoikis resistance model of AGS cells.Cell proliferation,apoptosis,invasion,migration,related proteins in PI3K/Akt signaling pathway,MMP-2 and SATB1 expression were detected to find differences with parental cells.MTT,FACS,scratch test,Transwell test and Western blot were used to investigate the effect of perifosine on biological behavior of anoikis resistant AGS cells.The effects of inhibition of signal transduction pathway and interference of SATB1 gene expression on the proliferation,apoptosis,invasion and migration of anoikis resistant AGS cells were studied.Results:Anoikis resistant AGS cell model was successfully constructed.Compared with the parental cells,the growth rate was faster,the apoptosis rate was lower,the migration and invasion ability were stronger,PIK3CA,Akt mRNA expression increased,PTEN mRNA expression decreased.P-Akt,SATB1,MMP-2 protein expression increased and PTEN protein expression decreased.Perifosine with different concentrations inhibited the growth of AGS cells,induced apoptosis,inhibited migration and invasion.The expression of p-Akt and MMP-2 protein decreased with the increase of perifosine concentration,while PTEN protein expression increased gradually.In addition,we found that perifosine decreased the expression of SATB1 protein in apoptosis resistant AGS cells,and the down-regulation of SATB1 enhanced the antitumor function of perifosine.Conclusion:Perifosine can play an anti-cancer role through PI3K/Akt pathway in apoptosis resistant AGS cells,and down-regulation of SATB1 expression can enhance this effect.Our study may provide a new potential anti-tumor treatment strategy for metastatic gastric cancer.

关 键 词：哌立福新 AGS PI3K/AKT信号通路 SATB1 失巢凋亡抵抗 

分 类 号：R735.2[医药卫生—肿瘤]