肾癌细胞系中HOTAIR表达的测定及稳定高表达HOTAIR肾癌细胞株的建立  

Determination of HOTAIR expression in renal cancer cell lines and establishment of a stable high expression HOTAIR kidney cancer cell line

作　　者：裴昌松[1,2] 范方田[3] 武艺[2] 沈存思[3] 潘立群[1] 

机构地区：[1]南京中医药大学第一临床学院,江苏南京210023 [2]徐州医学院第二附属医院泌尿外科,江苏徐州221006 [3]南京中医药大学药学院,江苏南京210023

出　　处：《南京医科大学学报（自然科学版）》2014年第5期548-552,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81373534);教育部博士点基金(20123237110003);江苏省"333工程"科研资助项目(BRA2013257)

摘　　要：目的:检测常用肾癌细胞系中HOTAIR的表达情况及建立稳定高表达HOTAIR肾癌细胞株。方法:①q-PCR检测正常肾细胞系(HK-2)和4种肾癌细胞系(os-rc-2,kevt-3,achn和769-p)中HOTAIR的表达;②利用携带HOTAIR基因的慢病毒(pLenti GFP Puro-HOTAIR)感染肾癌细胞系(769-p)后,并采用嘌呤霉素(Puro)抗性筛选方法建立高表达HOTAIR肾癌细胞株。结果:①5种细胞系HOTAIR表达的高低依次是kevt-3、os-rc-2、achn、769-p、HK-2;②慢病毒(pLenti GFP Puro-HOTAIR)转染肾癌细胞系(769-p)后,HOTAIR表达明显升高。结论:①高侵袭性肾癌细胞株(kevt-3、os-rc-2)中HOTAIR的表达明显高于低侵袭性肾癌细胞株(achn、769-p)及正常肾细胞株(HK-2);②成功建立稳定表达的HOTAIR的769-p-HOTAIR细胞系。Objective：To determine the expression of HOTAIR in common renal carcinoma and normal renal cell lines and establish a renal carcinoma cell line that highly expresses HOTAIR. Methods：①q-PCR was used to detect the expression of HOTAIR in four renal carcinoma cell lines （os-rc-2,kevt-3,achn and 769-p） and a normal renal cell line（HK-2）. ②Infection with lentiviral （pLenti GFP Puro-HOTAIR） carrying HOTAIR gene in 769-p cell line and puromycin-resistance screening were performed to establish the cell line that highly expresses HOTAIR. Results：① HOTAIR was expressed in five cell lines （from high to low）：kevt-3,os-rc-2,achn,769-p,and HK-2. ②HOTAIR was high expressed in 769-p after infected with lentiviral （pLenti GFP Puro-HOTAIR）. Conclusion：①HOTAIR expression in high metastatic carcinoma cell lines（kevt-3,os-rc-2） was higher than that in low metastatic renal carcinoma cell lines （achn,769-p ） and normal renal cell line（HK-2）.②The 769-p-HOTAIR cell line stably expressed HOTAIR.

关 键 词：HOTAIR 慢病毒载体 肾癌 

分 类 号：R737.11[医药卫生—肿瘤]