转录延伸样因子7对低分化人胃癌细胞株BGC823侵袭、迁移能力的影响及机制

Effect and mechanism of transcription elogation factor A （ SⅡ ） like 7 on invasion and migration of poorly differentiated human gastric cancer cell line BGC823

目的探讨转录延伸样因子7（TCEAL7）对低分化人胃癌细胞株BGC823侵袭、迁移的影响及其机制。 方法应用实时定量聚合酶链反应（Real-time PCR）和Western blot法检测TCELA7 mRNA和蛋白在BGC823和正常胃黏膜细胞株GES-1中的表达；构建TCEAL7真核表达载体GFP-TCEAL7并转染BGC823细胞48 h，检测转染效果；噻唑蓝（MTT）法检测GFP-TCEAL7转染对BGC823细胞活性的影响；划痕实验、Transwell小室实验检测细胞侵袭、迁移；应用Real-time PCR和Western blot法检测转染GFP-TCEAL7转染对BGC823细胞侵袭、迁移基因表达水平的影响。 结果Real-time PCR和Western blot法显示，BGC823的TCEAL7表达（mRNA：0.280±0.048，蛋白：0.181±0.038）明显低于GES-1（mRNA：0.474±0.062，蛋白：0.346±0.057；t＝－6.061，P＝0.000；t＝－5.900，P＝0.000）。GFP-TCEAL7转染后胃癌BGC823细胞TCEAL7的mRNA及蛋白表达（1.886±0.326、1.007±0.262）比对照组（0.302±0.053、0.172±0.030）显著上调（t＝11.748，P＝0.000；t＝7.756，P＝0.000）。MTT结果显示，转染了GFP-TCEAL7的BGC823细胞活性（0.672±0.083）比对照组（1.242±0.215）显著减弱（t＝－6.058，P＝0.000）。划痕实验、Transwell小室实验显示转染了GFP-TCEAL7的BGC823细胞迁移能力[转染组（53.945±9.952）%，对照组（81.017±7.208）%]和侵袭能力（转染组76.333±12.420，对照组121.167±11.771）均明显降低（t＝－5.396，P＝0.000；t＝－6.418，P＝0.000）。Real-time PCR和Western blot法显示转染GFP-TCEAL7的BGC823细胞基质金属蛋白酶（MMP）-7、MMP-9的mRNA和蛋白表达水平下降，组织基质金属蛋白酶抑制剂-1（TIMP1）和E-钙黏蛋白（E-cadherin）表达明显增高。 结论过表达TCEAL7可抑制BGC823细胞的侵袭、迁移。

ObjectiveTo explore the effects and mechanism of transcription elogation factor A （SⅡ） like 7 （TCEAL7） on invasion and migration of poorly differentiated human gastric cancer cell line BGC823 in vitro. MethodsReal-time quantitative polymerase chain reaction （Real-time PCR） and Western blotting were utilized to detect mRNA and protein expression of TCEAL7 in BGC823 cells and normal gastric mucosal cell line GES-1. TCEAL7 eukaryotic expression vector was designed and transfected into BGC823 cells for 48 h. The methyl thiazol tetrazolium （MTT） assay was applied to test the cell activity affected by GFP-TCEAL7 transfection. The wound assay and Transwell chamber assay were employed to detect the invasion and migration of cells. Real-time PCR and Western blotting were used to detect the mRNA and protein expression of invasion- and migration-related genes. ResultsReal-time PCR and Western blotting demonstrated that the expression of TCEAL7 was significantly wearker in BGC823 cells （mRNA： 0.280±0.048; protein： 0.181±0.038） than GES-1 cells （mRNA： 0.474±0.062; protein： 0.346±0.057; t=-6.061, P=0.000; t=-5.900, P=0.000）. The expression of TCEAL7 was up-regulated significantly in BGC823 cells after GFP-TCEAL7 transfection （1.886±0.326, 1.007±0.262） as compared with control group （0.302±0.053, 0.172±0.030; t=11.748, P=0.000; t=7.756, P=0.000）. Result of MTT exhibited that the proliferation inhibition rate increased significantly in BGC823 cells after GFP-TCEAL7 transfection （0.672±0.083） as compared with control group （1.242±0.215） （t=-6.058, P=0.000）. Wound assay and Transwell chamber assay revealed that GFP-TCEAL7 transfection induced decreased migration [for transfection group, （53.945±9.952）%; for control group, （81.017±7.208）%] and invasion [for transfection group, （76.333±12.420）; for control group, （121.167±11.771）] of cells （t=-5.396, P=0.000; t=-6.418, P=0.000）. The results of Real-time PCR and Western blotting showed that the mRNA and protein levels of matrix metalloproteinase （MMP）-7 and MMP-9 were significantly down-regulated by TCEAL7 overexpression, while the mRNA and protein expression levels of TIMP1 and E-cadherin up-regulated significantly. ConclusionOverexpression of TCEAL7 could inhibit invasion and migration of BGC823 cells.