稳定干扰ITGB1抑制人乳腺癌BT549细胞迁移和侵袭
发布时间：2018-07-29 15:42
【摘要】：在人乳腺癌细胞系BT549中稳定干扰整合素β1(integrinβ1,ITGB1),研究其对乳腺癌细胞迁移和侵袭的影响。构建靶向干扰ITGB1基因的sh RNA慢病毒,感染BT549细胞,通过筛选成功获得稳定干扰ITGB1蛋白的BT-549细胞系,实验设空白对照组(Blank)、病毒感染阴性对照组(shNC)、ITGB1-shRNA慢病毒感染组(shITGB1);Real-time PCR和Western blotting法检测稳定干扰后ITGB1 mRNA和蛋白的表达情况;Western blotting法检测稳定干扰ITGB1后上皮间质转化(epithelial-mesenchymal transition,EMT)相关标志蛋白(E-cadherin,Vimentin,N-cadherin,Fibronectin)的表达情况;利用划痕试实验和Transwell侵袭试实验分别检测稳定干扰ITGB1后细胞的迁移及侵袭能力。Real-time PCR和Western blotting结果显示,shITGB1组细胞ITGB1 mRNA和蛋白的表达水平显著低于Blank组和shNC组细胞(p0.01,p0.01)。Western blotting结果显示,稳定干扰ITGB1可部分逆转乳腺癌细胞BT549的EMT化;稳定干扰ITGB1后,上皮标志蛋白E-cadherin表达显著上调(p0.01),间质标志蛋白Vimentin(p0.05)、N-cadherin(p0.01)和Fibronectin(p0.01)的表达显著降低。划痕和Transwell侵袭试验实验结果显示,稳定干扰ITGB1后可显著降低乳腺癌细胞的迁移及侵袭能力(p0.05,p0.05)。在乳腺癌BT549细胞中稳定干扰ITGB1后,通过逆转EMT化抑制细胞迁移和侵袭。
[Abstract]:To study the effects of integrin 尾 1-ITGB1 on the migration and invasion of human breast cancer cell line BT549. Sh RNA lentivirus targeting ITGB1 gene was constructed and infected with BT549 cells. BT-549 cell lines with stable interfering ITGB1 protein were obtained by screening successfully. Real-time PCR and Western blotting methods were used to detect the expression of ITGB1 mRNA and protein after stable interference in the control group with negative (Blank), virus infection. Western blotting assay was used to detect epithelial-mesenchymal transition EMT related to stable interfering ITGB1. The expression of E-cadherin in N-cadherin fibronectin; The results of scratch test and Transwell invasion test showed that the expression of ITGB1 mRNA and protein was significantly lower than that of Blank group and shNC group (p0.01, p0.01). Western blotting showed that the expression level of ITGB1 mRNA and protein was significantly lower than that of Blank group and shNC group (p0.01). The results of real-time PCR and Western blotting showed that the expression of ITGB1 mRNA and protein was significantly lower than that of Blank group and shNC group (p0.01). Stable interference with ITGB1 could partially reverse the EMT expression of BT549 in breast cancer cells, and the expression of E-cadherin, Vimentin (p0.05), N-cadherin (p0.01) and Fibronectin (p0.01) were significantly up-regulated (p0.01), and the expression of Vimentin (p0.01) and Fibronectin (p0.01) were significantly decreased. The results of scratch and Transwell invasion tests showed that stable interference with ITGB1 could significantly reduce the migration and invasion ability of breast cancer cells (p0.05). Stable interference with ITGB1 in breast cancer BT549 cells inhibited migration and invasion by reversing EMT.
【作者单位】： 重庆医科大学检验医学院临床检验诊断学教育部重点实验室;成都市第三人民医院检验科;
【基金】：国家自然科学基金(31171336)项目资助
【分类号】：R737.9
[Abstract]:To study the effects of integrin 尾 1-ITGB1 on the migration and invasion of human breast cancer cell line BT549. Sh RNA lentivirus targeting ITGB1 gene was constructed and infected with BT549 cells. BT-549 cell lines with stable interfering ITGB1 protein were obtained by screening successfully. Real-time PCR and Western blotting methods were used to detect the expression of ITGB1 mRNA and protein after stable interference in the control group with negative (Blank), virus infection. Western blotting assay was used to detect epithelial-mesenchymal transition EMT related to stable interfering ITGB1. The expression of E-cadherin in N-cadherin fibronectin; The results of scratch test and Transwell invasion test showed that the expression of ITGB1 mRNA and protein was significantly lower than that of Blank group and shNC group (p0.01, p0.01). Western blotting showed that the expression level of ITGB1 mRNA and protein was significantly lower than that of Blank group and shNC group (p0.01). The results of real-time PCR and Western blotting showed that the expression of ITGB1 mRNA and protein was significantly lower than that of Blank group and shNC group (p0.01). Stable interference with ITGB1 could partially reverse the EMT expression of BT549 in breast cancer cells, and the expression of E-cadherin, Vimentin (p0.05), N-cadherin (p0.01) and Fibronectin (p0.01) were significantly up-regulated (p0.01), and the expression of Vimentin (p0.01) and Fibronectin (p0.01) were significantly decreased. The results of scratch and Transwell invasion tests showed that stable interference with ITGB1 could significantly reduce the migration and invasion ability of breast cancer cells (p0.05). Stable interference with ITGB1 in breast cancer BT549 cells inhibited migration and invasion by reversing EMT.
【作者单位】： 重庆医科大学检验医学院临床检验诊断学教育部重点实验室;成都市第三人民医院检验科;
【基金】：国家自然科学基金(31171336)项目资助
【分类号】：R737.9
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