与鼻咽癌细胞CNE-2L2恶性生物学行为相关的蛋白质  

Protein associated with malignant activities of human nasopharyngeal carcinoma cell CNE-2L2

作　　者：周异群[1] 时岩[1] 田云[1] 鞠吉雨[1] 陈双玲[1] 刘音[1] 朱立平[1] 

机构地区：[1]中国医学科学院中国协和医科大学基础医学研究所免疫学系,北京100005

出　　处：《基础医学与临床》2007年第4期364-371,共8页Basic and Clinical Medicine

基　　金：国家重点基础性研究项目(2001CB510004);国家高技术研究发展计划项目(2002BA711A01-17)

摘　　要：目的寻找与人鼻咽癌细胞CNE-2L2恶性生物学行为相关的蛋白质。方法用蛋白质组学分析高(W)和低(AS)恶性生物学行为的CNE-2L2细胞间蛋白质表达的差异,用实时定量PCR和免疫荧光染色流式细胞术对差异表达的蛋白质做确认,用siRNA抑制细胞中目的基因的表达,Western blot确认目的基因表达的抑制,细胞接种裸鼠的肿瘤生长实验。结果MOLDI-TOF MS分析见12个明显的差异表达蛋白质点。它们是:抗氧化蛋白Peroxiredoxin 2,异构型b;原肌蛋白3;II型TPM4-ALK融合癌蛋白;MLL蛋白;CD44;视网膜母细胞结合蛋白7;微管蛋白,β多肽;ATP合成酶,转H+作用的线粒体F1复合物,β多肽;波形蛋白;D21S2056E蛋白;蛋白酶体(prosome,macropain)26S subunit,non-ATPase,4;ENO1蛋白。前4个在AS细胞高表达,后8个在AS细胞低表达。对积分最高的7个蛋白质的差异表达做了肯定的验证。用siCD44抑制W细胞的CD44表达后,细胞接种裸鼠的成瘤性生长明显抑制。结论蛋白质组学分析见AS细胞与W细胞间有12个差异表达蛋白质,CD44表达减弱与AS细胞的恶性行为减弱相关。Objective To find genes associated with malignant activities of human nasopharyngeal carcinoma cell CNE-2L2. Methods Differential display of proteins between CNE-2L2 cells with high malignant activities （called W cell） and the CNE-2L2 ceils with profound reduced malignant activities （called AS cell） was analyzed by proteomic technique. The differential display was validated by real-time PCR or immune fluorescence staining. Expression of CD44 gene was inhibited by means of siRNA. The inhibition was validated by means of Western blotting. Results MOLDI-TOF MS showed 12 protein spots differential displayed. They were： Peroxiredoxin 2 isoform b;Tropomyosin 3; TPM4-ALK fusion oncoprotein type 2; MLL protein; CD44; Retinoblastoma binding protein 7; Tubulin, β polypeptide; ATP synthase, H ＋ transporting, mitochondrial F1 complex, β polypeptide; Vimentin; D21S2056E protein; Proteasome （ prosome, macropain） 26S subunit, non-ATPase ,4 ; ENO1 protein. The former 4 were up-regulated and the latter 8 were down-regulated in AS highest scores was confirmed. Tumorigenesis of the CNE-2L2 by siRNA in nude mice was profoundly inhibited. Conclusion found reduction of the malignant activities of CNE-2L2 cell. cells. The differential display of the 7 proteins with cells with inhibition of CD44 gene expression caused Inhibition of CD44 gene expression resultes in pro found reduction of the malignant activities of CNE-2L2 cell.

关 键 词：蛋白质组学 鼻咽癌细胞 恶性行为 CD44 

分 类 号：R739.63[医药卫生—肿瘤]