Optimization of transfection efficiency of small interfering RNA in purified human prolactinoma cells  

作　　者：SONG Yong-mei ZHAO Jiang YU Chun-jiang ZHAN Qi-min 

机构地区：[1]Department of Neurosurgery, Beijing Sanbo Brain Hospital of theCapital Medical University, Beijing 100093, China (Meng QH, [2]Department of Neurosurgery, Beijing Sanbo Brain Hospital of theCapital Medical University, Beijing 100093, China [3]State Key Laboratory of Molecular Oncology, Chinese Academy ofMedical Sciences and Peking Union Medical College, CancerInstitute and Cancer Hospital, Beijing 100021, China [4]Department of Neurosurgery, Daqing Oil-field General Hospital,Daqing, Heilongiiang 163001, China

出　　处：《Chinese Medical Journal》2011年第12期1862-1869,共8页中华医学杂志（英文版）

摘　　要：Background Control of hypersecretion of certain hormones is one of the key targets in the treatment of pituitary adenomas. RNA interference has been shown to inhibit protein expression, and thus it may represent a promising method for the treatment of pituitary adenomas. In the present study, transfection efficiency of small interfering RNA （siRNA） was optimized in human prolactinoma cells. Methods First, a method was optimized to extract highly purified human prolactinoma cells in vitro. The extracted cells were verified to retain the physiological features of prolactin （PRL） secretion. Second, three conditions for siRNA transfection were tested by the evaluation of transfectfon efficiency and cell viability. The proper transfection condition was verified for human prolactinoma cells. Third, the siRNA for prolactin was transfected into the human prolactinoma cells, and the suppression of PRL mRNA was evaluated by quantitative real-time reverse transcription-PCR. Results The siRNA of 100 pmol with Lipofectamine 2000 of 5 μl for 1×10^6 cells was proved preferable, with transfection efficiency being 53.3% and cell viability being 69.7%. In the preliminary experiment the siRNA against PRL decreased the mRNA of PRL by 34.0%. Conclusion It is possible to inhibit hormone hypersecretion by RNA interference, that may eventually enable therapeutic siRNA drugs developed.Background Control of hypersecretion of certain hormones is one of the key targets in the treatment of pituitary adenomas. RNA interference has been shown to inhibit protein expression, and thus it may represent a promising method for the treatment of pituitary adenomas. In the present study, transfection efficiency of small interfering RNA （siRNA） was optimized in human prolactinoma cells. Methods First, a method was optimized to extract highly purified human prolactinoma cells in vitro. The extracted cells were verified to retain the physiological features of prolactin （PRL） secretion. Second, three conditions for siRNA transfection were tested by the evaluation of transfectfon efficiency and cell viability. The proper transfection condition was verified for human prolactinoma cells. Third, the siRNA for prolactin was transfected into the human prolactinoma cells, and the suppression of PRL mRNA was evaluated by quantitative real-time reverse transcription-PCR. Results The siRNA of 100 pmol with Lipofectamine 2000 of 5 μl for 1×10^6 cells was proved preferable, with transfection efficiency being 53.3% and cell viability being 69.7%. In the preliminary experiment the siRNA against PRL decreased the mRNA of PRL by 34.0%. Conclusion It is possible to inhibit hormone hypersecretion by RNA interference, that may eventually enable therapeutic siRNA drugs developed.

关 键 词：RNA interference PROLACTINOMA PROLACTIN cell culture techniques  transfection 

分 类 号：F[经济管理]