敲低心肌连接斑株蛋白对人胃癌SGC-7901细胞的迁移和侵袭能力的影响及机制研究

Effects of JUP on Migration and Invasion Ability of Human Gastric Cancer Cell Line SGC-7901 and Its Mechanism

该文研究了敲低心肌连接斑株蛋白(junction plakoglobin,JUP)对人胃癌SGC-7901细胞迁移和侵袭的影响及其机制。通过sh RNA慢病毒介导的方式构建了敲低JUP基因的慢病毒载体序列及其阴性对照病毒,并感染至人胃癌SGC-7901细胞中,荧光检测细胞感染效率,获得JUP基因稳定沉默及其对照细胞株。用嘌呤霉素筛选阳性转染细胞株,采用实时荧光定量PCR(q RT-PCR)和蛋白质免疫印记法(Western blot)检测JUP表达水平。采用细胞划痕愈合实验和Transwell小室实验检测敲低JUP基因之后对人胃癌SGC-7901细胞迁移和侵袭能力的影响。采用Western blot检测敲低JUP之后β-连环蛋白水平的影响。结果显示,通过sh RNA介导的慢病毒成功构建了敲低JUP基因的人胃癌SGC-7901稳定细胞株。与对照组相比,敲低JUP表达可促进细胞的迁移和侵袭能力(P<0.05),上调了β-连环蛋白的水平(P<0.05)。以上结果表明,敲低人胃癌SGC-7901细胞中JUP基因表达后可促进胃癌细胞SGC-7901的迁移和侵袭能力可能与其上调β-连环蛋白的水平从而活化Wnt信号通路有关。

The purpose of this study was to investigate the effects of junction plakoglobin （JUP） knockdown on the migration and invasion of human gastric cancer cell line SGC-7901 and its mechanism. The lentiviral vector sequence of knockdown JUP gene and its negative control virus were constructed by shRNA lentivirus-mediated method. The lentivirus and its negative control virus were transfected into human gastric cancer cell SGC-7901. After transfection for 3 days, the transfection efficiency of the cells was detected by microscope fluorescence, finally get well silenced JUP gene cell and the control. Positive transfected cell lines were screened with puromycin, the Real-time fluorescence quantitative PCR and Western blot were used to detect the JUP mRNA and protein level. The migration and invasion ability of human gastric cancer SGC-7901 cells were detected by cell wound healing and Transwell chamber test. Western blot was used to detect the effect of JUP knockdown on the level of β-catenin. The results showed that human gastric cancer SGC-7901 stable cell line knockdown by JUP gene was successfully constructed by shRNA-mediated lentivirus. Compared with control, knockdown JUP gene could promote cell migration and invasion （P〈0.05）, the level of β-catenin was increased （P〈0.05）. The above results indicated that knockdown of JUP gene expression in human gastric cancer SGC-7901 cells might promote the migration and invasion of gastric cancer cell SGC-7901, which might be related to its upregulation β-catenin and activation of Wnt signaling pathway.