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
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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)


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

    Abstract:

    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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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(自然科学版),2014,(5):548-552

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  • 收稿日期:2013-12-29
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  • 在线发布日期: 2014-05-21
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