摘要
目的研究调控血小板反应蛋白1(TSP1)的表达对宫颈癌HeLa细胞增殖、迁移、凋亡、侵袭及促进内皮细胞成血管能力的影响。方法构建表达TSP1基因的真核表达载体pcDNA3.1-TSP1及siRNA-TSP1,转染宫颈癌HeLa细胞,实验共分为TSP1基因过表达组(pcDNA3.1-TSP1转染组)、TSP1基因低表达组(siRNA-TSP1转染组)和未转染组(阴性对照组)。Real-time PCR、Western blot检测各组细胞TSP1的表达,CCK-8、划痕实验、流式细胞术、Transwell检测各组细胞生长情况,体外血管形成实验检测各组细胞促血管生成能力。结果与阴性对照组比较,pcDNA3.1-TSP1组细胞TSP1 mRNA及蛋白表达上调,siRNA-TSP1组细胞TSP1 mRNA及蛋白表达下调,差异均有统计学意义(P<0.05);pcDNA3.1-TSP1组HeLa细胞的增殖、迁移、侵袭及体外促内皮细胞血管形成能力明显增强,差异均有统计学意义(P<0.05),但凋亡比例比较差异无统计学意义(P>0.05),而siRNA-TSP1组HeLa细胞的增殖、迁移、侵袭及体外促内皮细胞血管形成能力均降低,凋亡比则增加,差异均有统计学意义(P<0.05)。结论下调TSP1的表达可抑制HeLa细胞的增殖、迁移、侵袭及促进内皮细胞成血管能力,促进细胞凋亡。上调TSP1则可促进He La细胞的增殖、迁移、侵袭及体外促血管形成能力,但对细胞凋亡无明显影响。
Objective To investigate the effect of regulating thrombospondin 1(TSP1)expression on the proliferation,migration,apoptosis,invasion of cervical cancer HeLa cells and its ability to promote endothelial cell angiogenesis.Methods Eukaryotic expression vectors pcDNA3.1-TSP1 and siRNA-TSP1 were constructed.They were used to transfected the cervical cancer HeLa cells.The experiment was divided into TSP1 gene overexpression group(pcDNA3.1-TSP1 transfection group),TSP1 gene under-expression group(siRNA-TSP1 transfection group),and non-transfection group(control group).Real-time PCR and Western blot were used to detect the expression of TSP1 in each group.CCK-8,wound healing assay,flow cytometry,Transwell assay,and matrigel assay were used to determine the changes of biological behavior of each group.Results Compared with the control group,the expression of TSP1 mRNA and protein was up-regulated in pcDNA3.1-TSP1 group and down-regulated in siRNA-TSP1 group(P<0.05).Compared with the control group,the proliferation,migration,and invasion of HeLa cells and the ability to promote endothelial cell angiogenesis in vitro were significantly enhanced in pcDNA3.1-TSP1 group(P<0.05);but there was no significant difference in the proportion of apoptosis between the two groups(P>0.05).In the siRNA-TSP1 group,the proliferation,migration,and invasion of HeLa cells,and the ability to promote endothelial cell angiogenesis in vitro were significantly decreased compared with those in the control group,while apoptosis was increased,with statistically significant differences(P<0.05).Conclusion Down-regulation of TSP1 expression can inhibit the proliferation,migration and invasion of HeLa cells,inhibit the ability to promote endothelial cell angiogenesis in vitro,and promote cell apoptosis.Up-regulation of TSP1 expression could promote the proliferation,migration and invasion of HeLa cells,improve the ability to promote endothelial cell angiogenesis in vitro,but has little effect on the apoptosis of HeLa cells.
作者
张花
潘莹
何全中
ZHANG Hua;PAN Ying;HE Quan-zhong(Department of Obstetrics and Gynecology,the Third Hospital of Xinxiang Medical University,Xinxiang 453000,Henan,CHINA)
出处
《海南医学》
CAS
2023年第6期768-773,共6页
Hainan Medical Journal
