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利用RNA干扰技术研究PTX1在鼻咽癌中的功能
PTX1 in nasopharyngeal carcinoma by RNAi technology
【摘要】 目的:探讨位于鼻咽癌高频扩增区+12p12-p11的PTX1基因在鼻咽癌中的表达及生物学功能。方法:用RT-PCR和荧光定量RT-PCR检测PTX1在36例鼻咽癌及8例慢性鼻咽炎中的表达。构建发夹状pSUPER-shPTX1干扰载体,转染鼻咽癌细胞系6-10B。mRNA水平检测PTX1基因的干扰效果,采用细胞周期及细胞凋亡检测PTX1被干扰后对鼻咽癌细胞6-10B细胞生物学特性的影响。结果:RT-PCR和荧光定量RT-PCR显示PTX1在鼻咽癌中高表达(P<0.05)。RNA干扰PTX1能抑制鼻咽癌细胞系的细胞增殖,诱导凋亡。结论:RNAi对PTX1的表达阻断改变了鼻咽癌细胞系6-10B的生物学特性,提示鼻咽癌12p12-p11扩增区获得的PTX1基因可能通过促进细胞的增殖和减少凋亡双途径来参与鼻咽癌的发生发展。
【Abstract】 ObjectiveTo explore the expression and the role of PTX1 located at the amplified 12p12-p11 region in nasopharyngeal carcinoma (NPC). MethodsSemi-quantitative RT-PCR and real-time RT-PCR were applied to detect the expression level of PTX1 in 36 NPC and 8 chronic nasopharyngitis (NP) biopsies. RNAi vector targeting PTX1 was constructed and transfected into NPC cell line 6-10B. The RNAi effect was determined by detecting the expression level of PTX1 in transfected 6-10B cell line. Finally, the cell biological characteristics were compared between transfected 6-10B and parental 6-10B by analyzing the cell cycle distribution and apoptosis status using flow cytometry. ResultsRT-PCR and real-time RT-PCR revealed that PTX1 gene was over-expressed in NPC tissues (P<0.05). PTX1 expression was suppressed in NPC cell line 6-10B by approximately 65% by RNAi, confirmed by RT-PCR. The depletion of PTX1 could effectively block the proliferation and induce the apoptosis of NPC cells. ConclusionBlocking the expression of PTX1 on mRNA level changed the characterization of NPC cell line 6-10B by RNAi, suggesting that PTX1 identified in the amplified 12p12-p11 region may be involved in the genesis and development of NPC via promoting the cell proliferation and inhibiting the cell apoptosis.
【Key words】 nasopharyngeal carcinoma; +12p12- p11; PTX1; RNA interference;
- 【文献出处】 中南大学学报(医学版) ,Journal of Central South University(Medical Sciences) , 编辑部邮箱 ,2007年02期
- 【分类号】R739.63
- 【被引频次】1
- 【下载频次】145