文章摘要
裴昌松,范方田,武 艺,沈存思,潘立群.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[J].南京医科大学学报,2014,(5):548~552
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
投稿时间:2013-12-29  
DOI:10.7655/NYDXBNS20140503
中文关键词: HOTAIR  慢病毒载体  肾癌
英文关键词: HOTAIR  lentiviral vector  renal cell cancer
基金项目: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)
作者单位
裴昌松 南京中医药大学第一临床学院,江苏 南京 210023
徐州医学院第二附属医院泌尿外科,江苏 徐州 221006 
范方田 南京中医药大学药学院,江苏 南京 210023 
武 艺 徐州医学院第二附属医院泌尿外科,江苏 徐州 221006 
沈存思 南京中医药大学药学院,江苏 南京 210023 
潘立群 南京中医药大学第一临床学院,江苏 南京 210023 
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中文摘要:
      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?
英文摘要:
      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.
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