肿瘤相关新基因pp4519在肺癌中的表达差异及初步功能研究  

Differential expression and preliminary function study of a novel tumor related gene pp4519

作　　者：罗金红[1] 李锦军[1] 张锋锐[1] 陈燕[2] 冯强[3] 覃文新[1] 万大方[1] 顾健人[1] 

机构地区：[1]上海交通大学肿瘤研究所 [2]中科院上海生物工程中心,上海200233 [3]浙江大学医学院附属第一医院胸外科,杭州310003

出　　处：《肿瘤》2004年第3期201-208,共8页Tumor

基　　金：国家863项目(编号 :2 0 0 1AA2 2 712 1;2 0 0 1AA2 2 114 1);国家十五攻关重大项目(编号:2 0 0 3BA711A0 2 1);国家 973重大项目(编号 :2 0 0 2CB5 13 10 0 4);上海市科委重大基础研究课题 (编号:0 3DJ14 0 0 7)

摘　　要：目的　探讨肿瘤相关新基因 pp4 5 19在肺癌细胞系、人肺癌及癌旁组织中表达差异及其初步生物学功能。 方法　合成肿瘤相关新基因pp4 5 19特异性多肽 ,免疫家兔制备兔源性多克隆抗体 ;从体外细胞集落形成试验 ,生长曲线 ,裸鼠体内成瘤试验 ,半定量RT PCR ,蛋白印迹 ,免疫细胞化学及免疫组化检测 pp4 5 19基因的表达差异 ,以及转染肺癌细胞系的细胞凋亡检测等方法研究 pp4 5 19基因对体内外细胞生长和生物学功能的影响。 结果　SPC A1肺癌细胞系的体外集落形成试验表明 ,pp4 5 19基因转染组集落形成数明显少于空载体组 (P <0 .0 5 ) ;转染 pp4 5 19基因的肺癌细胞系的裸鼠体内成瘤试验结果表明该基因对SPC A1细胞系的成瘤有明显抑制作用 (P <0 .0 1) ;瞬时转染细胞的流式细胞凋亡检测结果表明该基因具有促进SPC A1细胞凋亡的作用 ;半定量RT PCR ,蛋白印迹检测结果表明肺癌及癌旁组织中pp4 5 19mRNA和蛋白表达无明显差异(P >0 .0 5 ) ,但免疫组化结果显示PP4 5 19在人肺癌癌旁组织中的表达略高于肺癌。结论　pp4 5Objective To explore the differential expression of a novel tumor related gene pp4519 in lung cancer lines, human lung cancer and noncancerous lung tissues as well as its preliminary biological function. Methods Specific oligopeptide (22 AAs) of PP4519 was synthesized and conjugated to KLH, polyclonal rabbit anti PP4519 antibody was prepared. The biological function and differential expression of PP4519 were studied by in vitro colony formation assay, growth curve of transfected cells, ectopic tumor formation assay of transfected SPC A1 cell line, semi Q RT PCR, western blot, immunocytochemistry immunohistochemistry, and so forth. Results Compared to vector, PP4519 inhibited the colony formation of transfected SPC A1 cells in vitro ( P <0.05); PP4519 also inhibited significantly the ectopic tumor formation of transfected SPC A1 cells in nude Balb/C mice ( P <0.01). Apoptotic ratio both of SPC A1 and COS 7 cells were increased by transiently transfecting pp4519 gene. No significant different expression was observed in lung cancer and noncancerous lung tissues as measured by semi Q RT PCR, western blot( P >0.05, P >0.05). However, the result of immunohistochemistry showed that PP4519 was up expressed in noncancerous lung tissues than normal lung tissues. Conclusion pp4519 was a novel lung cancer related gene with the function of inhibiting its growth.

关 键 词：肿瘤相关基因 pp4519基因 肺癌 肿瘤生物学 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]