摘要
目的探讨Krüppel样转录因子8(KLF8)在骨肉瘤中的表达及对骨肉瘤细胞生物学特性的影响。方法通过RT-PCR和Western blot法分别检测骨肉瘤组织及配对的正常非肿瘤组织中KLF8 mRNA和蛋白水平。以骨肉瘤细胞F5M2为研究对象,细胞转染KLF8小干扰RNA(KLF8 siRNA组)及对照阴性序列(siRNA control组),同时以不做处理的细胞为对照组。RT-PCR和Western blot检测转染后细胞中KLF8 mRNA和蛋白水平。噻唑蓝(MTT)检测细胞增殖,流式细胞术检测细胞凋亡,Western blot检测细胞中信号转导与转录因子3(STAT3)、磷酸化的STAT3(pSTAT3)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)表达。结果 KLF8在骨肉瘤组织中的表达水平明显高于非肿瘤组织(P<0.05)。siRNA control组细胞中KLF8、STAT3、p-STAT3、Cleaved Caspase-3水平及细胞存活率、凋亡率与对照组相比无显著差异(P>0.05)。KLF8 siRNA组细胞存活率及细胞中p-STAT3、KLF8水平均明显低于对照组(P<0.05)。KLF8 siRNA组细胞凋亡率及细胞中Cleaved Caspase-3水平均明显高于对照组(P<0.05)。结论 KLF8在骨肉瘤组织中表达上调。干扰KLF8表达可能通过作用于STAT3信号通路抑制骨肉瘤细胞增殖,促进骨肉瘤细胞凋亡。
Objective To study on the Krüppel-like factor 8（ KLF8） expression in osteosarcoma and its effect on osteosarcoma cell biological characteristics. Methods The levels of KLF8 mRNA and protein were detected by RT-PCR and Western blot in osteosarcoma tissues and paired with normal non tumor tissues. Osteosarcoma cell F5 M2 was used as the research object,cells were transfected with KLF8 small interfering RNA（ KLF8 siRNA group） and negative control（ siRNA control group）,at the same time,set up the control group which was not treated. Detection of KLF8 mRNA and protein levels in transfected cells by RT-PCR and Western blot. Cell proliferation was detected by MTT,cell apoptosis was detected by flow cytometry,Western blot was used to detect the expression of STAT3,p-STAT3 and Cleaved Caspase-3 in cysteine. Results The expression level of KLF8 in osteosarcoma was significantly higher than that in non tumor tissues（ P〈0. 05）. The levels of KLF8,STAT3,p-STAT3,Cleaved Caspase-3 and cell survival rate and apoptosis rate in siRNA control group were not significantly different from those in control group（ P〈0. 05）. The cell survival rate and p-STAT3 and KLF8 levels in KLF8 siRNA group were significantly lower than those in control group（ P〈0. 05）. The apoptosis rate and Cleaved Caspase-3 levels in KLF8 siRNA group were significantly higher than those in control group（ P〈0. 05）. Conclusion Up regulation of KLF8 expression in osteosarcoma. Interference of KLF8 expression may inhibit the proliferation of osteosarcoma cells and promote apoptosis of osteosarcoma cells by acting on STAT3 signaling pathway.
作者
冯庆虎
王友良
FENG Qing-hu;WANG You-liang(Department of Orthopeadic Surgery,The First People＇s Hospital ofLiangshan Yi Autonomous Prefecture,Xichang Sichuan 615000,China.)
出处
《临床和实验医学杂志》
2018年第17期1851-1855,共5页
Journal of Clinical and Experimental Medicine
