| 促凋亡基因PDCD5在骨肉瘤U2-OS细胞表达及对其促凋亡作用的研究 |
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| 引用本文： | 王海斌,孙永明,胥正权,田康松. 促凋亡基因PDCD5在骨肉瘤U2-OS细胞表达及对其促凋亡作用的研究[J]. 中华肿瘤防治杂志, 2010, 17(4) |
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| 作者姓名： | 王海斌 孙永明 胥正权 田康松 |
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| 作者单位： | 苏州大学附属第二医院骨科,江苏,苏州,215004 |
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| 摘 要： | 目的:观察外源性PDCD5基因对骨肉瘤细胞系U2-OS的促凋亡作用。方法:扩增重组质粒PCI-neo-PDCD5,采用脂质体转染的方法,将PDCD5基因转染骨肉瘤细胞U2-OS,RT-PCR检测转染后表达情况;撤除血清培养16h后,四甲基偶氮唑盐MTT法检测目的基因转染后细胞的增殖情况;An-nexinV-染色流式细胞仪检测细胞凋亡;蛋白质印迹法分析凋亡通路。结果:转染骨肉瘤细胞U2-OS后,RT-PCR检测到PDCD5的表达,MTT实验发现撤除血清后转染重组质粒组细胞的增殖被明显抑制。于转染空载体和对照组细胞相比,存活细胞明显减少,P<0.001。AnnexinV染色,流式细胞仪检测可见明显的凋亡细胞,与转染空质粒组(凋亡率为1.47%)相比,其凋亡率为10.28%。经PDCD5处理的细胞中,Bcl-2、Caspase-3和Caspase-9的表达与对照组比较,差异均有统计学意义。结论:PDCD5基因可以在转染的骨肉瘤细胞系U2-OS细胞中表达,并在撤出血清因素的诱导作用下可显著抑制转染细胞的生长,并通过线粒体激活途径发挥促凋亡作用并诱导细胞发生凋亡。
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| 关 键 词： | 骨肉瘤/病理学 基因 PDCD5 细胞凋亡 流式细胞术 转染 |
Expression of PDCD5 and human Osteosarcoma U2-OS cells |
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| WANG Hai-bin,SUN Yong-ming,XU Zheng-quan,TIAN Kang-song. Expression of PDCD5 and human Osteosarcoma U2-OS cells[J]. Chinese Journal of Cancer Prevention and Treatment, 2010, 17(4) |
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| Authors: | WANG Hai-bin SUN Yong-ming XU Zheng-quan TIAN Kang-song |
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| Affiliation: | WANG+Hai-bin,SUN+Yong-ming,XU+Zheng-quan,TIAN+Kang-song+Department+of+Orthopedic,Second+Affiliated+Hospital+of+Soochow+University,Suzhou+215004,P.R.China |
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| Abstract: | OBJECTIVE:To investigate the apoptosis-promoting effect of PDCD5 on human osteosarcoma U2-OS cells.METHODS:PCI-neo and PCI-neo-PDCD5 were transfected into U2-OS cells by Lipofectamine 2000,The expression of PDCD5 mRNA was detected by RT-PCR.The viability of the cells was analyzed by MTT assay and Annexin V-FITC staining 16 hours after the removal of the serum.The apoptosis pathway was analysised by Western blot.RESULTS:After osteosarcoma U2-OS cells transfected with PDCD5 gene,the target protein was express... |
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| Keywords: | osteosarcoma/pathology genes PDCD5 apoptosis flow cytometry transfection |
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