慢病毒介导的Notch1基因沉默抑制软骨肉瘤SW1353细胞的增殖和侵袭

Inhibitory effect of lentivirus mediated Notch1 gene silencing on proliferation and invasion of chondrosarcoma SW1353 cells

目的:探讨慢病毒介导的Notch1基因沉默对软骨肉瘤SW1353细胞增殖和侵袭的抑制作用及其机制。方法:将SW1353细胞分为对照组(未感染)、空载体组(LV-NC-shRNA慢病毒感染)和沉默组(LV-Notch1-shRNA慢病毒感染),RT-PCR和Western blot检测慢病毒的感染效果,MTT法和克隆形成实验检测各组细胞的增殖活力,Transwell小室实验检测细胞的侵袭能力,Western blot检测PCNA、MMP-2和p-AKT蛋白的表达。结果:LV-Notch1-shRNA慢病毒感染使SW1353细胞中Notch1 mRNA和Notch1、PCNA、MMP-2、p-AKT蛋白的表达均明显降低(P<0.05),显著抑制了SW1353细胞的增殖和侵袭能力(P<0.05)。结论:Notch1基因沉默可抑制软骨肉瘤SW1353细胞增殖和侵袭,其作用机制可能与下调PCNA、MMP-2和p-AKT蛋白表达有关。

Objective:To investigate the inhibitory effect of lentivirus-mediated Notch1 gene silencing on the proliferation and invasion of chondrosarcoma SW1353 cells and its mechanism.Methods:SW1353 cells were divided into control group (uninfected),blank load group (LV-NC-shRNA lentiviral infection) and silent group (LV-Notch1-shRNA lentiviral infection).The infection effect of lentivirus was detected by RT-PCR and Western blot.The proliferative activity of cells in each group was measured by MTT assay and clone formation test.Cell invasiveness was tested by Transwell cell assay,and the expressions of PCNA,MMP-2 and p-AKT proteins were measured by Western blot.Results:The expressions of Notch1 mRNA and Notch1,MMP-2,PCNA and p-AKT proteins in SW1353 cells were significantly decreased,and the proliferation and invasion ability of SW1353 cells were inhibited by LV-Notch1-shRNA lentiviral infection.Conclusion:Notch1 gene silencing can inhibit the proliferation of chondrosarcoma SW1353 cells,and its mechanism may be related to down-regulation of PCNA,MMP-2 and p-AKT protein expression.