质粒转染对HEK293和DDT1-MF2细胞天然β2-肾上腺素受体表达的影响

Effects of plasmid transfection on native β_2-adrenoceptor expression in HEK293 and DDT1-MF2 cell lines

目的 :观察质粒转染对细胞天然 β2 肾上腺素受体 (β2 AR)表达量的影响。方法 :将不携带外源目的基因的 pREP4质粒和携带α1B AR或 β1 ARcDNA的 pREP4质粒 ,采用脂质体和磷酸钙沉淀法转染HEK2 93和DDT1 MF2细胞。结果 :相同质粒载体 ,无论有无外源目的基因 ,或目的基因是否相同 ,均可使HEK2 93细胞天然表达的β2 AR密度及其介导的cAMP蓄积效应增加。而用同样方法在DDT1 MF2细胞转染 pREP4质粒则对 β2 AR表达无影响。结论 :转染携带或不携带目的基因的质粒对HEK2 93细胞天然表达 β2 AR的密度及其介导的功能效应均有影响 ,且提示这种效应可能具有细胞特异性。

Objective: To investigate the effects of plasmid transfection on native β 2 adrenoceptor (AR) expression in cell lines. Methods: The pREP4 plasmids with adrenoceptor cDNAs or without exogenous cDNA were transfected into humam embrynonic kindney 293 (HEK293) and DDT1 MF2 cell lines by calcium phosphate co precipitation and lipofection mediated transfection methods. Results: The densities of native β 2 AR and the cAMP accumulation induced by β 2 AR were significantly increased in HEK293 cells transfected with pREP4, whether the AR cDNAs were inserted into pREP4 plasmids or the exogenous cDNAs were the same. However, no such effect was found in DDT1 MF2 cells. Conclusion: These results suggest that plasmid transfection may be responsible for the increase of the native β 2 AR expression and its mediated cAMP accumulation in HEK293 cells, and this effect might be cell type related.