RNA干扰介导的转录因子E2F-1沉默对人胃癌裸鼠移植瘤生长的影响  

The effects of RNA interference of E2F transcription factor 1 on human gastric subcutaneous tumor of nude mice

作　　者：王晓通[1] 钱倩[1] 李雷[1] 谢玉波[2] 肖强[1] 

机构地区：[1]广西医科大学第一附属医院胃肠腺体外科,南宁530021 [2]广西医科大学第一附属医院麻醉科,南宁530021

出　　处：《中华实验外科杂志》2011年第3期340-342,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：国家自然科学基金资助项目（30860273）;桂林科技攻关计划资助项目（10124001A-22）

摘　　要：目的 观察转录因子E2F-1基因RNA干扰（RNAi）的重组慢病毒载体（pLL-E2F-1-shRNA）对人胃癌裸鼠移植瘤E2F-1基因的表达和肿瘤生长的影响.方法 建立人胃癌裸鼠移植瘤模型,随机分为pLL-E2F-1-shRNA组,pLL3.7组,生理盐水组.隔天向各组分别注射相应药物0.2 ml,共6次,并测量各组肿瘤体积.11 d后处死裸鼠,显微镜观察移植瘤组织形态变化;免疫组织化学法、半定量逆转录-聚合酶链反应（RT-PCR）和Western blot检测移植瘤中E2F-1基因的表达,脱氧核苷酸末端转移酶介导的缺口末端标记（TUNEL）检测肿瘤细胞凋亡指数.结果 与生理盐水组和pLL3.7组比较,pLL-E2F-1-shRNA组肿瘤生长速度明显减慢（P＜0.05）;病理学观察确认所取组织为恶性肿瘤组织;免疫组织化学结果显示pLL3.7组和生理盐水组的肿瘤细胞细胞核内有少量棕褐色颗粒沉着,pLL-E2F-1-shRNA组肿瘤细胞细胞核内均未见或偶见棕褐色颗粒沉着;半定量RT-PCR和Western blot结果显示pLL-E2F-1-shRNA组E2F-1 mRNA表达较pLL3.7组和生理盐水组明显下降（P＜0.05）,E2F-1蛋白表达较pLL3.7组和生理盐水组减少75.2%和83.9%;pLL-E2F-1-shRNA组的凋亡指数（16.7±5.6）%明显高于pLL3.7组（10.5±4.1）%和生理盐水组（11.2±4.3）%（P＜0.05）.结论 慢病毒载体pLL-E2F-1-shRNA瘤内注射可抑制人胃癌裸鼠移植瘤的生长.Objective To study the effects of pre-built recombinant lentiviral vector for RNA interference （RNAi） of E2F transcription factor 1 （E2F-1） gene （pLL-E2F-1-shRNA） on the expression of E2F-1 gene and the growth of human gastric subcutaneous tumor of nude mice.Methods BALB/C nude mice were inoculated subcutaneously with MGC-803 cells to establish the nude mouse gastric cell carcinoma model,and they were randomly divided into pLL-E2F-1-shRNA group,pLL3.7 group and NS group.The corresponding agents were injected （0.2 ml per time） for consecutive 6 times every two days,and the tumor volume was measured at the same time.Nude mice were killed after 11 days.After HE staining,histopathological characteristics of subcutaneous tumor were observed under a microscope.The expression of E2F-1 was detected by using immunohistochemistry,semi-quantitative reverse transcription polymerase chain reaction （RT-PCR） and Western blotting respectively.The apoptosis of tumor cells was assayed by using in situ end labeling technique （TUNEL）.Results The mean relative tumor volume （RTV） of tumor growth in the pLL-E2F-1-shRNA group was significantly lower than that in the pLL3.7 group and NS group （ P〈0.05 ）.The tissue obtained from the nude mice was malignant tumor under the microscopy after HE staining.Immunohistochemistry revealed that there were a few of brown granules in pLL3.7 group and NS group,while there were almost no brown granules in the pLL-E2F-1-shRNA group.The expression level of E2F-1 mRNA in pLL-E2F-1-shRNA group was lower than in pLL3.7 group and NS group.As compared with pLL3.7 group and NS group （ P〈0.05 ）,E2F-1 protein in pLL-E2F-1-shRNA group was decreased by 75.2% and 83.9% respectively.The apoptosis rate in pLL-E2F-1-shRNA group was （ 16.7 ±5.6）%,significantly higher than that in other two groups [（ 10.5 ±4.1 ）% for pLL3.7 group and （ 11.2 ±4.3）% for NS group （P 〈0.05）].Conclusion Intra-tumor injection of retrovirus pLL-E2F-1-shRNA has significantly inhibitor

关 键 词：慢病毒载体 RNA干扰 E2F-1 胃癌 

分 类 号：R735.2[医药卫生—肿瘤]