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研究了稳定表达hERG基因的CHO-K1(中国仓鼠卵巢细胞)细胞株及其在药物研发安全性筛选方面的应用。使用Lipofectamine 2000系统将hERG基因转染入CHO-K1细胞,用Western Blot检测其表达,并进一步通过FluxORTM Thallium Assay Kits验证;同时使用FluxORTM Thallium Assay Kits检测Cisapride和Dofetilide对hERG通道的抑制性。建立了4株稳定表达hERG基因的CHO-K1细胞株,Cisapride和Dofetilide对hERG通道的IC50分别为86.7 nmol.L-1和28.1 nmol.L-1。hERG基因成功地在CHO-K1细胞株实现了稳定表达,此细胞株可用于FluxORTM Thallium Assay Kits系统,用于药物研发安全性筛选,以评估化合物潜在的心脏毒性。
The CHO-K1 (Chinese hamster ovary cell) cell strain stably expressing hERG gene was studied and its application in the safety screening of drug development. The hERG gene was transfected into CHO-K1 cells using the Lipofectamine 2000 system. The expression of hERG was detected by Western Blot and further verified by FluxORTM Thallium Assay Kits. The inhibitory effect of Cisapride and Dofetilide on hERG channel was also tested by FluxORTM Thallium Assay Kits. Four CHO-K1 cell lines stably expressing hERG gene were established. The IC50 of Cisapride and Dofetilide for hERG channel were 86.7 nmol.L-1 and 28.1 nmol.L-1, respectively. The hERG gene was successfully expressed in the CHO-K1 cell line and was used in the FluxORTM Thallium Assay Kits system for safety screening of drug development to assess potential cardiotoxicity of the compound.