TAGLN基因过表达对乳腺癌细胞迁移和侵袭能力的影响

目的 研究TAGLN基因过表达对人乳腺癌细胞MDA-MB-231迁移能力和侵袭能力的影响及其分子机制.方法 对MDA-MB-231细胞采用慢病毒表达系统构建TAGLN基因稳定过表达细胞株,将MDA-MB-231细胞正常培养设为空白对照组,空载体慢病毒包装感染MDA-MB-231细胞后获得的稳定转染细胞株设为空载体对照组.Real time PCR和Western blot检测TAGLN mRNA和蛋白表达,划痕实验和Transwell侵袭实验检测细胞迁移和侵袭能力,Western blot检测TAGLN基因过表达后基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)和MMP-9蛋白表达变化.结果 MDA-MB-231细胞感染TAGLN基因过表达慢病毒载体后,细胞中TAGLN mRNA表达和蛋白表达升高(均P＜0.01),成功构建TAGLN基因过表达稳定细胞株(231-TAGLN).231-TAGLN细胞的体外迁移能力和侵袭能力下降,与空载体对照组和空白对照组细胞比较,差异均具有统计学意义(均P＜0.01),同时伴有MMP-2和MMP-9表达水平降低(均P＜0.01).结论 TAGLN基因过表达可以抑制乳腺癌细胞的迁移和侵袭,MMP-2和MMP-9基因表达下降可能参与这一过程.

Effects of TAGLN overexpression on the migration and invasion of breast cancer cell

Objective To investigate the effects of TAGLN overexpression on the migration and invasion of human breast cancer cell MDA-MB-231 and the molecular mechanism.Methods Lentiviral expression system was used in MDA-MB-231 cells to construct the TAGLN stable overexpression cell lines.MDA-MB-231 cells untreated were used as blank control group and those stably transfected with empty vector lentivirus were used as control group.The expression of TAGLN mRNA and protein were detected by real time PCR and Western blot.Wound scratch assay and transwell invasion assay were carried out to determine the migration and invasion ability of cells.Western blot was applied to determine the expression levels of matrix metalloproteinase-2 (MMP-2) and MMP-9 after TAGLN overexpression.Results After the infection of TAGLN overexpression lentivirus vector in MDA-MB-231 cells,the expression of TAGLN mRNA and protein were signi ficantly increased (both P ＜ 0.01),which suggested the successful construction of TAGLN stable overexpression cell lines (231-TAGLN).The in vitro migration and invasion ability of 231-TAGLN cells were decreased significantly,compared with the control group and blank control group (all P ＜ 0.01).Moreover,the expression levels of MMP-2 and MMP-9 were both decreased in 231-TAGLN cells (both P ＜ 0.01).Conclusion TAGLN overexpression significantly inhibits the migration and invasion of breast cancer cells,and the downregulation of MMP-2 and MMP-9 expression may be involved in this process.