Tat结合蛋白30对胃癌细胞SGC7901细胞侵袭转移的影响
Effect of Tat interactive protein 30 expression on migration and invasion of gastric cancer cell line SGC7901
目的 检测Tat结合蛋白30((Tat interactive protein 30,TIP30)在胃癌组织中的表达水平及其过表达后对胃癌细胞SGC7901侵袭转移的影响.方法 采用免疫组化方法检测93例胃癌组织中TIP30的表达;采用pcDNA3.1-TIP30质粒转染SGC7901细胞,通过MTT法检测SGC7901细胞活性的变化;Transwell小室实验检测SGC7901细胞中TIP30过表达后侵袭、迁移能力的变化;采用Western blot检测在TIP30过表达SGC7901细胞中E-cadherin、N-cadherin和基质金属蛋白酶-9表达的改变.结果 TIP30蛋白在胃癌组织中表达明显低于正常胃组织(39%比92%),差异有统计学意义(x2 =32.68,P<0.05);TIP30的表达与胃癌浸润深度、淋巴结转移、TNM分期均有关(x2=3.535、7.421、6.754,均P<0.05);与对照组比较,72 h和96 hTIP30转染组细胞增殖水平明显降低(t=6.528、7.249,均P<0.05);TIP30过表达组胃癌细胞体外迁移及侵袭能力均显著减弱(=5.769,P<0.05;t =7.886,P<0.05);TIP30过表达组细胞中基质金属蛋白酶-9和N-cadherin蛋白表达明显降低(t=9.811、10.362,均P<0.05),而E-cadherin蛋白表达水平明显增加(t=6.137,P<0.05).结论 TIP30在胃癌组织中低表达,上调其表达后可以抑制胃癌SGC7901细胞增殖活性和侵袭迁移能力.
更多Objective To investigate the expression of TIP 30 protein in gastric cancer tissues,and effect of TIP30 over-expression on migration and invasion of gastric cancer cell line SGC7901.Methods Immunohistochemistry streptavid-in-peroxidase (SP) methods were used to detect the expression levels of TIP30 in 93 cases of gastric cancer tissues.Previously constructed pcDNA3.1-TIP30 plasmid were transiently transfected into SGC7901 cells.The proliferation of cells were detected by using MTT assay when TIP30 was overexpressed.Transwell assay to determine migration and invasion ability of SGC7901.Western blot was used to examine the changes of concentration of E-cadherin,N-cadherin and MMP9.Results The positive expression rate of TIP30 was 39% significantly lower in gastric cancer tissues than 92% in normal gastric mucosa tissues (x2 =32.68,P < 0.05),there was a significant correlation between reduced expression of TIP30 and depth of infiltration,including nodal metastasis,TNM stage (x2 =3.535,7.421,6.754,all P < 0.05);MTT showed that the proliferation of SGC7901 cells in the pcDNA3.1-TIP30 transfected group significantly decreased when TIP30 was overexpressed at respective time of 72,96 hours (t =6.528,7.249,both P < 0.05),Transwell assay showed that overexpression of TIP30 significantly decreased migratory and invasive numbers of SGC7901 cells (t =5.769,P < 0.05;t =7.886,P < 0.05);the expression level of MMP-9 and N-cadherin in TIP30 overexpressing cells group significantly decreased (t =9.811,10.362,both P < 0.05),mean while E-cadherin expression was significantly higher than before (t =6.137,P < 0.05).Conclusion TIP30 protein is low expressed in gastric cancer and the overexpression of TIP30 inhibits the proliferation,migration and invasion of gastric cell line SGC7901.
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