人新基因ASB-8 SOCS盒缺失突变体抑制肺癌细胞生长  

Exogenous Expression of SOCS Box-deficient Mutant ASB-8 Suppresses the Growth of Lung Adenocarcinoma SPC-A1 Cells

作　　者：刘永忠[1] 李锦军[1] 张锋锐[1] 沈雁[2] 姚明[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,癌基因及相关基因国家重点实验室,上海200032 [2]中国科学院上海生物工程研究中心,上海200233

出　　处：《生物化学与生物物理学报》2003年第6期548-553,共6页

基　　金：国家重点基础研究发展规划项目 ( 973计划 ) (No .G19980 5 12 0 9);国家高技术研究发展计划 ( 863计划 )资助项目(No .2 0 0 1AA2 2 712 1和No .2 0 0 1AA2 2 114 1)~~

摘　　要：ASB 8是新近发现的含锚蛋白重复序列和细胞因子信号抑制物盒蛋白质家族新成员 (humanankyrinre peatandSOCSboxcontainingproteinfamily ,ASBfamily)。研究观察了人ASB 8在肺癌中的表达及对肿瘤细胞生长的影响。利用制备的ASB 8多克隆抗体和免疫组化技术 ,分析了人ASB 8在肺癌组织中的表达 ;构建ASB 8及其SOCS盒缺失突变体 (ASB 8ΔSB)的真核表达载体 ,转染至人肺腺癌细胞系SPC A1中并检测外源基因的表达 ;通过体外细胞生长曲线、裸鼠体内成瘤试验 ,观察ASB 8及其缺失突变体对SPC A1细胞生长的影响。免疫组化结果表明 :ASB 8蛋白在肺癌组织中高表达 ,阳性率为 96 .8% (30 / 31) ,而在癌旁肺和正常肺组织低表达或不表达 ,弱阳性率仅为 2 5 .8% (8/ 31) ;转染ASB 8ΔSB的SPC A1肺癌细胞的增殖速度显著慢于对照组细胞 (P <0 .0 1) ,而转染ASB 8的肺癌细胞没有明显的生长特性改变 ;ASB 8SOCS盒缺失突变体对SPC A1的体内生长及肿瘤形成有一定的抑制作用 (P <0 .0 1)。ASB 8基因可能是一种与人肺癌发生发展相关的新基因。ASB-8 is a new member of the human ankyrin repeat and SOCS box containing protein family (ASB). This report deals with the expression of ASB-8 protein in lung carcinoma tissue, as well as its biological effect on the proliferation and growth of lung cancer cell line SPC-A1. ASB -8 was expressed in pT7-450 expression vector, and an anti-ASB-8 rabbit polyclonal antibody was prepared. The expression of ASB-8 protein in lung carcinoma tissue was detected using immunohistochemistry. The growth characteristics of the lung adenocarcinoma SPC-A1 cells expressing either exogenous ASB-8 or ASB-8ΔSB (SOCS box-deficient) was studied by both in vitro cell growth curve and in vivo nude mouse tumor formation assay. Immunohistochemical staining detected positive reaction of 96.8% (30/31) showing that ASB-8 was highly expressed in lung cancer tissues; however, the expression of ASB-8 was lower, or even no expression in noncancerous lung tissues. Significant growth inhibition was observed in SPC-A1 cell line expressing ASB-8ΔSB on day 4 and persisted to day 6, compared with mock transfected cells ( P <0.01). No difference in the growth properties was observed between ASB-8 and mock transfected cells. In vivo study also showed that tumor formation of ASB-8ΔSB expressing cells was significantly inhibited as compared with that of the control group ( P <0.01). Therefore, ASB-8 may play an important role in growth and proliferation of lung cancer, possibly as positive regulator.

关 键 词：人 基因ASB 8SOCS盒 缺失突变体 肺癌 细胞因子 

分 类 号：R734.2[医药卫生—肿瘤]