摘要
目的探讨沉默MAT1基因表达对骨肉瘤细胞增殖与迁移的影响,以及对SDF1-CXCR4信号通路的作用。方法qRT-PCR检测人成骨细胞hFOB1.19与骨肉瘤细胞株Saos-2、MG63、U2OS、143B中MAT1 mRNA的表达水平。将对数生长期143B细胞分为空白对照组(不进行转染)、阴性对照组(转染阴性对照siRNA-NC)和siRNA-MAT1组(转染siRNA-MAT1),通过CCK-8、流式细胞术及Transwell小室法分别检测各组143B细胞增殖活力、细胞周期和迁移能力,Western blot法检测各组143B细胞中SDF1和CXCR4蛋白表达情况。结果与hFOB1.19细胞比较,骨肉瘤细胞株Saos-2、MG63、U2OS中MAT1 mRNA的表达水平明显升高(P<0.01),143B细胞中MAT1 mRNA的表达水平显著升高(P=0.000);siRNA-MAT1组143B细胞中MAT1 mRNA的表达水平较空白对照组(Blank组)和siRNA-NC组明显下降(P<0.01);与Blank组和siRNA-NC组比较,siRNA-MAT1组143B细胞增殖力明显降低,G 1期比例升高而S期比例降低,细胞迁移能力受到抑制,SDF1和CXCR4蛋白表达水平也明显下降(P<0.01)。结论沉默FOSL2基因的表达可抑制骨肉瘤细胞143B的增殖和迁移能力,机制可能与下调SDF1和CXCR4表达有关。
Objective To investigate the effect of silencing MAT1 gene expression on proliferation and migration of osteosarcoma cells and the effect of silencing MAT1 gene on SDF1-CXCR4 signaling pathway.Methods qRT-PCR was used to detect the expression level of MAT1 mRNA in human osteoblast hFOB1.19 and osteosarcoma cell lines Saos-2,MG63,U2OS and 143B.Logarithmic growth phase 143B cells were divided into blank control group(no transfection),negative control group(transfection negative control siRNA-NC)and siRNA-MAT1 group(transfection siRNA-MAT1).The proliferation,cell cycle and migration ability of 143B cells in each group were detected by CCK-8,flow cytometry and Transwell chamber method.Western blot was used to detect the expression of SDF1 and CXCR4 in 143B cells.Results Compared with hFOB1.19 cells,the expression levels of MAT1 mRNA in osteosarcoma cell lines Saos-2,MG63 and U2OS were significantly increased(P<0.01),and the expression level of MAT1 mRNA in 143B cells was significantly increased(P=0.000).The expression level of MAT1 mRNA in siRNA-MAT1 group was significantly lower than that in the blank control group(Blank group)and siRNA-NC group(P<0.01).Compared with the Blank group and the siRNA-NC group,the proliferation of 143B cells in the siRNA-MAT1 group was significantly decreased,the proportion of G 1 phase was increased,the proportion of S phase was decreased,the cell migration ability was inhibited,and the expression levels of SDF1 and CXCR4 proteins were also significantly decreased(P<0.01).Conclusion Silencing FOSL2 gene expression can inhibit the proliferation and migration of osteosarcoma cell line 143B,and the mechanism may be related to down-regulation of SDF1 and CXCR4 expression.
作者
巨啸晨
舒莉
周玲
孙荣鑫
Ju Xiaochen;Shu Li;Zhou Ling(Department of Joint Surgery,The Sixth Affiliated Hospital of Xinjiang Medical University,Xinjiang 830000,China)
出处
《医学研究杂志》
2020年第5期88-92,共5页
Journal of Medical Research
基金
新疆维吾尔自治区自然科学基金资助项目(2017D01C257)。
