RON变异体RONΔ170对自身酪氨酸磷酸化变异体RONΔ160的显性抑制作用  

Dominant negative effect of RONΔ170,a splicing product of RON receptor tyrosine kinase,on an auto-phosphorylating variant RONΔ160

作　　者：邓晶[1,2] 张敷彪[1] 劳伟峰[1] 王达[1] 黄学锋[1] 

机构地区：[1]浙江大学医学院附属邵逸夫医院肛肠外科,浙江省生物治疗重点实验室,浙江杭州310016 [2]浙江医学高等专科学校

出　　处：《全科医学临床与教育》2009年第5期498-500,F0002,共4页Clinical Education of General Practice

基　　金：浙江大学医学院中青年科研启动基金资助项目(491030-542927)

摘　　要：目的研究受体型酪氨酸激酶RON的变异体RONΔ170对RON的自身酪氨酸磷酸化变异体RONΔ160的作用。方法NIH-3T3和3T3-RONΔ160细胞转染质粒pcDNA3.1-RONΔ170,建立3T3-RONΔ170和3T3-RONΔ160Δ170细胞株;流式细胞仪检测RONΔ170蛋白的表达;免疫沉淀法检测RON的酪氨酸磷酸化;软琼脂培养法观察RONΔ170对3T3和3T3-RONΔ160细胞生长能力的作用。结果成功建立3T3-RONΔ170和3T3-RONΔ160Δ170细胞株,流式细胞仪胞膜蛋白检测法证明其稳定表达RONΔ170蛋白;免疫沉淀试验结果显示RONΔ170能明显抑制RONΔ160的RON酪氨酸磷酸化;软琼脂生长实验示3T3、3T3-RONΔ160、3T3-RONΔ170和3T3-RONΔ160Δ170细胞生长克隆数比较,差异有统计学意义(t=7.93,P<0.05)。结论受体型酪氨酸激酶RON的变异体RONΔ170无酪氨酸磷酸化功能,且具有显性抑制变异体特性。Objective To investigate the roles of RON△170 receptor tyrosine kinase on an auto-phosphorylating variant RON△160 in 3T3 cell line. Method 3T3-RON△170 and 3T3-RON△160RON△170 cell lines were constructed; the tyrosine phosphorylation of RON was measured by immunoprecipitation method; Cell growth and motile ability were tested by soft agar assay. Results 3T3-RON△170 and 3T3-RON△160RON△170 cell lines were constructed successfully; 3T3-RON△170 had no phosphorylated RON expression; phosphorylated RON expression of 3T3-RON△160RON△170 cell was lower than 3T3-RON△160. Soft agar results showed growth cell clones of 3T3, 3T3-RON△160, 3T3-RON△170 and 3T3 -RON△160RON△170 were 12.33 ±2.08,121.33 ±12.58,14.00 ±3.00 and 34.33±8.32; 3T3-RON△160 compared to 3T3-RON△16ORON△170, t=7.93,P〈0.05. Conclusions RON△170 is a potential dominant negative variant of RON receptor tyrosine kinase.

关 键 词：蛋白质体酪氨酸激酶 细胞生长 肿瘤浸润 显性抑制 

分 类 号：R341[医药卫生—基础医学]