LYN对胃癌生物学行为的影响及调控机制的研究  

LYN effects on the biological behavior of gastric cancer and its regulatory mechanisms

作　　者：苏锐[1] 路艳[2] 郑鑫[1] Su Rui;Lu Yan;Zheng Xin(Department of Gastrointestinal Surgery,Affiliated Hospital of Chengde Medical College,Chengde,067000,China;Department of Anesthesiology,Affiliated Hospital of Chengde Medical College,Chengde 067000,China)

机构地区：[1]承德医学院附属医院胃肠外科,067000 [2]承德医学院附属医院麻醉科,067000

出　　处：《中华实验外科杂志》2020年第12期2232-2235,共4页Chinese Journal of Experimental Surgery

基　　金：承德市科学技术研究与发展计划项目(201904A104)。

摘　　要：目的观察LYN对胃癌细胞迁移、侵袭等生物学行为的影响并分析其调控机制。方法2018年2月至2019年12月,通过蛋白印迹分析(Western blot)检测在承德医学院附属医院2017年至2018年间接受外科手术患者的LYN,并临床收集的胃癌和癌旁组织样本中的表达差异;免疫组化检测分析LYN与临床病理指标间的关系;利用RNA干扰技术敲低LYN在胃癌细胞系AGS中的表达,Western blot检测转染效率;克隆形成和Transwell实验分别用于检测LYN敲低对AGS细胞增殖、迁移和侵袭的影响;Western blot检测相关信号通路相关蛋白;使用信号途径激活剂IGF-1进行拯救实验,分析IGF-1是否可以逆转LYN敲低对AGS细胞增殖、迁移和侵袭的抑制作用,对其调控机制进行研究。两组比较采用独立样本t检验。结果LYN在胃癌组织中的的表达被显著上调;LYN在胃癌组织中的表达与患者的病理分级及分期显著相关;sh-LYN转染胃癌细胞可使LYN蛋白的表达量低于正常对照组(P<0.05);转染后,sh-LYN组的细胞克隆形成率为(25.2±1.8)%,细胞侵袭数目为(51.67±3.53)个,迁移细胞数目为(231.70±11.98)个,与NC组比较,sh-LYN组细胞的增殖、迁移、侵袭受到抑制(t=3.850、3.437、4.450,P<0.05);Western blot检测结果,与正常对照组比较,sh-LYN敲低LYN的表达之后,可显著降低Wnt3和β-catenin的表达(t=7.058、5.617,P<0.05),而且IGF-1逆转了LYN敲低对AGS细胞增殖、迁移和侵袭的抑制作用。结论敲低LYN可抑制胃癌细胞的增殖、迁移和侵袭,并诱导胃癌细胞凋亡,其过程与Wnt/β-catenin信号通路有关。Objective To study the effects of LYN on biological behaviors such as proliferation,migration and invasion of gastric cancer cells,and to analyze their regulatory mechanisms.Methods This study was conducted from February 2018 to December 2019.Western blotting was used to detect expression differences in LYN’s gastric cancer and para-cancer tissue samples collected from our hospital during surgery between 2017 and 2018.Immunohistochemical analysis of the relationship between LYN and clinicopathological indicators;RNA interference was used to knock down LYN expression in GASTRIC cancer cell line AGS,and Western blotting was used to detect transfection efficiency.The effects of LYN knockdown on the proliferation,migration and invasion of AGS cells were examined by cloning and Transwell experiments.Western blotting was used to detect proteins associated with relevant signal pathways.Rescue experiments were carried out using igF-1,a signaling pathway activator,to analyze whether IGF-1 could reverse the inhibitory effect of LYN knockdown on AGS cell proliferation,migration and invasion,and to investigate its regulatory mechanism.Results LYN expression was significantly upregulated in gastric cancer.LYN expression in gastric cancer tissue was significantly correlated with the patient’s pathological grade and stage.The expression of LYN protein in AGS cells after SH-lyn transfection was significantly lower than that of the normal control group(NC)(t=4.816,P<0.05);After transfection,the cell clonal formation rate of sh-lyn group was(25.2±1.8)%,the number of cell invasion was(51.67±3.53),and the number of migrating cells was(231.70±11.98).Compared with the NC group,the proliferation,migration and invasion of gastric cancer cells in sh-lyn group were inhibited(t=3.850,3.437,4.450,P<0.05);Western blotting results showed that the expression of Wnt3 andβ-catenin in sh-lyn group was significantly lower than that in NC group(t=7.058,5.617,P<0.05),in addition,IGF-1 reversed the inhibition of LYN knockdown on AGS cell prol

关 键 词：LYN 胃癌 迁移 侵袭 WNT/Β-CATENIN信号通路 

分 类 号：R735.2[医药卫生—肿瘤]