摘要
目的 观察结肠癌转移相关基因-1（MACC-1）表达对体外胃癌细胞株MGC-803的增殖凋亡和生物学功能的影响.方法 培养胃癌MGC-803细胞,将传代的细胞分为以下3组：A组：MACC-1-短发卡RNA （shRNA）和PSV2neo3共转染组,B组：shNC和PSV2neo共转染组,C组：未转染组,A组为实验组,B、C两组为阳性对照组和阴性对照组.通过Western blot和实时定量聚合酶链反应（Real-time PCR）检测MACC-1蛋白和mRNA在3组细胞中的表达水平,并检测3组MGC-803细胞中酪氨酸激酶受体（c-Met）、E-钙黏蛋白（E-cadherin）和N-钙黏蛋白（N-cadherin）和mRNA水平表达变化,并通过流式细胞法和划痕实验观察比较转染MACC-1-shRNA的MGC-803细胞的的迁移能力和增殖凋亡情况.结果 （1）Westem blot和Real-time PCR检测结果显示：与B、C组比较,MACC-1-shRNA组MACC-1蛋白及mRNA表达均显著下调（P＜0.01）,c-Met蛋白及其mRNA的表达显著下调（P＜0.01）.（2）Western blot和Real-time PCR检测结果显示：与B、C组比较,A组上皮标志物E-cadherin蛋白和及mRNA表达显著上调（P＜0.01）,c-Met蛋白及其mRNA,间质细胞标志物N-cadherin蛋白及mRNA的表达显著下调（P＜0.01）,B、C两组间E-cadherin和N-cadherin蛋白及mRNA表达的差异无统计学意义（P＞0.01）.（3）划痕实验检测细胞迁移：结果表明,A组划痕间距较宽,愈合被抑制,MGC-803细胞迁移能力低于B、C细胞（P＜0.01）,B、C两组细胞的划痕间距差异无统计学意义（P＞0.01）.（4）流式细胞仪分析细胞凋亡：A、B和C组细胞的凋亡率分别为（21.06＆#177;1.12）％、（2.18＆#177;0.14）％和（2.04＆#177;0.11）％,A组细胞的凋亡率较C组明显增加（P＜0.01）,B、C两组细胞凋亡的差异无统计学意义.结论 MACC-1基因转染对体外胃癌细胞株MGC-803细胞具有促进增殖和抑制凋亡作用,其可能是通过调控肝细胞生长因子（HGF）/c-Met信号转导通路来发挥作用.
Objective To study the effect of colon cancer metastasis associated gene-1 （MACC-1） expression on the proliferation and apoptosis of gastric cancer cell line MGC-803 in vitro.Methods Cultured gastric cancer MGC-803 ceils and passaged cells were divided into three groups：group A [MACC-1-short hairpin RNA （shRNA） and PSV2neo3 co transfection group];group B （shNC and PSV2neo co transfection group）;group C （non transfection group）.By Western blotting and real-time quantitative polymerase chain reaction （Real-time PCR）,the expression of MACC-1 protein and mRNA was detected in the three groups,and that of of c-Met,E-cadherin and N-cadherin in 3 groups of MGC-803 cells were detected.Flow cytometry and scratch test were used to analyze migration,proliferation and apotosis of MGC-803 cells transfected with MACC-1-shRNA.Results （1） Western blotting and Real-time PCR test results showed that as compared with groups B and C,MACC-1 and c-Met protein and mRNA expression in group A were significantly down-regulated （P 〈 0.01）.（2） Western blotting and Real-time PCR detection results showed as compared with groups B and C,E-cadherin protein and mRNA expression levels were significantly increased （P 〈 0.01）,c-Met and N-cadherin mRNA and protein levels were significantly decreased （P 〈 0.01） in group A.There was no significant difference between groups B and C.（3） Scratch assay showed the scratch space was wide,and healing was inhibited in group A.the migration ability of MGC-803 cells in group A was reduced as compared with groups B and C （P 〈 0.01）,but there was no statistically significant difference in the scratch space between groups B and C.（4） Flow cytometry revealed that the apoptosis rate in groups A,B and C was （21.06 ＆#177; 1.12）%,（2.18 ＆#177;0.14）% and （2.04 ＆#177; 0.11%） respectively.The apoptosis rate in group A was significantly higher than in group C （P 〈 0.01）,and there was no significant difference between groups B and C.Conclusi
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第5期1230-1233,共4页
Chinese Journal of Experimental Surgery
关键词
胃癌
结肠癌转移相关基因-1
脱噬作用
生物学功能
Gastric cancer
Colon cancer metastasis associated gene - 1
Apoptosis
Biological function
