摘要
目的探讨锌指蛋白ZNF139在骨肉瘤细胞和组织中表达,及对骨肉瘤细胞143B增殖和转移的影响,并初步探讨其作用机制。方法通过qRT-PCR和免疫蛋白印迹实验分析ZNF139在骨肉瘤细胞系和组织中的相对表达;转染pcDNA3.1(+)-ZNF139质粒(实验组)和pcDNA3.1(+)-Vector质粒(对照组)至143B细胞系中,免疫蛋白印迹实验验证ZNF139的过表达;通过CCK-8实验和细胞克隆实验分析ZNF139对细胞增殖的影响;transwell细胞迁移和侵袭实验分析ZNF139对细胞迁移和侵袭的影响;流式细胞周期和凋亡实验验证ZNF139过表达对细胞增殖的作用机制;免疫蛋白印迹实验验证ZNF139基因对p53信号通路的影响。结果与正常成骨细胞和组织相比,ZNF139在骨肉瘤细胞和组织中的表达明显下调(P<0.001),差异具有统计学意义;与对照组相比(100%),转染后24、48、72 h细胞活性受到明显的抑制(P<0.05),实验组细胞克隆形成明显受到抑制(43.44%±5.55%)(P<0.05);过表达ZNF139后细胞迁移和侵袭能力明显受到抑制(P<0.001);流式细胞实验发现,实验组细胞明显阻滞于G1期(P<0.001),同时实验组中细胞凋亡率明显升高(P<0.001);过表达ZNF139后通过促进p53的表达及下游信号转导。结论 ZNF139在人骨肉瘤瘤组织中表达下调,通过促进p53信号通路参与骨肉瘤的进展。
Objective To explore the expression of ZNF139 in osteosarcoma cell lines and tissues, and the effects of ZNF139 on proliferation and migration of osteosarcoma cell lines 143 B. Methods qRT-PCR and western blot(WB) method were used to analysis the relative expression of ZNF139 in osteosarcoma cell lines and tissues. ZNF139 was transfect into 143 B cell, ectopic expression of ZNF139 was measured by WB. To investigate the function of ZNF139, CCK-8 and cell proliferation were used, utilized flow cytometric analyses of the cell cycle and apoptosis, each experiment was repeated three times independently, and the average was statistically analyzed. And also used transwell experiment to analysis the ability of cell migration and invation. qRT-PCR and WB were used to analysis the basic mechanism of its tumor-suppressitive ability. Results ZNF139 mRNA and proteins were downregulated in osteosarcoma cell lines and tissues, cell viability and colony formation in ectopic expression of ZNF139 in osteosarcoma cell ZNF139 cells were significantly inhibited comparing to control group(P<0.001), also transwell migration and invation assay showed that ectopic expression of ZNF139 in 143 B cell lines resulted in a significant decrease in cell than control group. Furthermore, ZNF139 restrained cell proliferation and invation through inducing p53 and its downstream target genes accumulation. Conclusion ZNF139 appears to be a functional tumor suppressor regulating tumorigenesis of osteosarcoma by Stabilizing p53 signaling.
作者
谢易
李彬彬
龚泰芳
Xie Yi;Li Binbin;Gong Taifang(Department of Orthopaedics,Taihe Hospital Affiliated to Hubei University of Medicine,Shiyan 442000,Hubei,China)
出处
《贵州医药》
CAS
2019年第11期1688-1692,共5页
Guizhou Medical Journal
关键词
骨肉瘤
ZNF139
抑癌基因
增殖和转移
P53
Osteosarcoma
ZNF139
Tumor suppressor genes
Cell proliferation/migration
p53
