siRNA干扰人胰腺癌T3细胞系KAI1基因的表达  

The expression of KAII gene by small interfering RNA （siRNA） in human pancreatic cancer cell line T3

作　　者：弭延斌[1] 郭晓钟[2] 刘峰[2] 徐建华[2] 田宏[2] 夏春莲[2] 吴开春[1] 樊代明[1] 

机构地区：[1]第四军医大学附属西京医院消化内科,西安710032 [2]沈阳军区总医院消化内科

出　　处：《中华胰腺病杂志》2008年第2期81-83,共3页Chinese Journal of Pancreatology

基　　金：国家自然科学基金（30670594）

摘　　要：目的应用小分子干扰RNA（small interference RNA，siRNA）干扰KAI1基因在人胰腺癌T3细胞系的表达，为基因治疗胰腺癌打下基础。方法针对KAI1基因CD82片段序列设计A、B、C、D4个siRNA靶序列，构建针对KAI1基因（CD82）含siRNA片段的慢病毒载体。以脂质体2000转染T3细胞，测定病毒滴度。以空载体、含不同靶序列siRNAd1载体的病毒感染T3细胞（MOI=5），采用实时PCR方法检测CD82mRNA的表达。结果正常对照组、空载体对照组、A靶序列组、B靶序列组、C靶序列组和D靶序列组CD82mRNA表达量分别为1．398±0．242、1．311±0．048、0．664±0．093、0．345±0．032、0．641±0．049和0．147±0．049，各靶序列组的表达量明显低于空载体对照组（P〈0．01）。结论府用RNAi可有效抑制r13细胞KAI1基因CD82片段的表达。Objective To evaluate the expression of KAI1 （ CD82 ） gene inhibited by small interfering RNA （siRNA） in human pancreatic cancer cell line T3. Methods Four sequences of siRNA including A, B, C, D were designed, which were based on the KAI1 gene sequence using online RNA interfering designing software and lentivirus vector was built. Then they were used to transfect T3 cells by liposome 2000 and virus titer was determined. Empty vector containing siRNAdl lentivrus particle （ MOI = 5 ） was also used to infect T3 cells. The expression of CDs2 mRNA was detected by real-time PCR. Results The expression of CD82 mRNA in normal control group, empty vector group, A group, B group, C group, D group were 1. 398 ± 0. 242, 1. 311 ± 0.048, 0. 664 ± 0. 093, 0. 345 ± 0. 032, 0. 641 ± 0. 049 and 0. 147 ± 0. 049, respectively, the difference between the expression of CDs2 mRNA in empty vector group and that of A, B, C, D groups was significant （P 〈0.01 ）. Conclusions RNAi was able to inhibit the expression of KAI1 gene CD82 in human pancreatic cancer cell line T3.

关 键 词：胰腺肿瘤 RNA 小分子干扰 KAI1基因 细胞系 

分 类 号：R735.9[医药卫生—肿瘤] R394[医药卫生—临床医学]