胰腺癌细胞BXPC-3转染条件的优化

Optimization of transfection conditions for human pancreatic cancer cell line BXPC-3

目的探讨实验室常用的人胰腺癌细胞株BXPC-3的有效转染方法及最佳转染参数。方法通过脂质体法和电穿孔法将表达绿色荧光蛋白的质粒(p GFP-N3)导入人胰腺癌细胞株BXPC-3中,24 h后观察并比较细胞的存活率及瞬转效率,确定最佳转染参数。结果常规脂质体法BXPC-3细胞瞬转阳性率为1.36%,细胞存活率为87.45%;而电穿孔法在电压120 V电击时程70μs条件下,BXPC-3细胞瞬转阳性率为54.78%,细胞存活率为64.01%,2种转染方法的瞬转阳性率差异有统计学意义(P<0.05)。结论转染BXPC-3细胞,电穿孔法可取得比常规脂质体法更高的转染效率。通过条件的优化,电穿孔法可以用于某些脂质体法难以有效转染的细胞株。

Objective To investigate transfection methods and conditions for human pancreatic cancer cell line BXPC -3 and related optimal transfection data .Methods The plasm pGFP-N3 was transfected into BXPC-3 cells respectively by lipofectamine 2000 and electroporation .Then, survival and transient transfection rates of the cells were observed and compared after 24 hours, so as to i-dentify the optimal transfection data .Results The positive BXPC-3 transient transfection efficiency by lipofectamine 2 000 was 1.36%, with the cell survival rate of 87.45%, while the transient transfection efficiency by electroporation with the pulse length of 70μs and e-lectric shock of 120 V was 54.78%, with the cell survival rate of 64.01%.Statistics indicated that there were significant differences in the transient transfection rates, when comparisons were made between the 2 transient transfection methods（P〈0.05）.Conclusion In the transfection of BXPC-3, electroporation could achieve higher transfection rates than the conventional lipofectamine .Through optimi-zation of transfection conditions , electroporation could effectively transfect certain cell strains , which were hard to be transfected by lipo-fectamine.