Effect and mechanism of transcription elogation factor A (SII) like 7 on invasion and migration of poorly differentiated human gastric cancer cell line BGC823
Wei Liang, Wei Geng, Yanan Fan, Zhibin Ye, Ming Gao
Abstract
Wei Liang, Wei Geng, Yanan Fan, Zhibin Ye, Ming Gao
Abstract
Objective To 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. Methods Real-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. Results Real-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. Conclusion Overexpression of TCEAL7 could inhibit invasion and migration of BGC823 cells. Key words: Gastric neoplasms; Transcription elogation factor A (SⅡ) like 7; Invasion; Migration; Gene cloning
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Objective To 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. Methods Real-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. Results Real-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. Conclusion Overexpression of TCEAL7 could inhibit invasion and migration of BGC823 cells. Key words: Gastric neoplasms; Transcription elogation factor A (SⅡ) like 7; Invasion; Migration; Gene cloning
Key concepts: Transfection, Molecular biology, Blot, Cell culture, Gentamicin protection assay, MTT assay, Messenger RNA, Cell