转基因法建立膀胱癌多药耐药细胞株

Establishment of multidrug resistance bladder cancer cell line by gene transfection

目的:建立高效、稳定的人膀胱癌多药耐药性细胞模型。方法:采用基因重组技术构建真核表达载体pcDNA3.1(+)/bcl-2,通过脂质体将人bcl-2基因转染膀胱癌细胞BIU-87,RT-PCR检测基因转录水平,应用MTT比色法进行耐药性检验。结果:酶切鉴定和基因测序证明真核表达载体pcDNA3.1(+)/bcl-2构建成功;RT-PCR分析表明,转染bcl-2基因的BIU-87/bcl-2细胞的bcl-2基因表达水平较BIU-87细胞和转染空载体的BIU-87/neo细胞显著提高;与BIU-87细胞和BIU-87/neo细胞相比,BIU-87/bcl-2细胞具有较高的耐药性。结论:基因转染法是建立人膀胱癌多药耐药细胞模型的理想方法。

Objective：To establish highly effective and stable muhidrug resistance （MDR）bladder cancer cell line. Methods ： Gene recombination was performed to construct the eucaryotic expression vector pcDNA3.1 （＋）/bcl-2. The establishment of the vector was verified by enzyme incision and gene sequencing. The human bladder cancer BIU-87 cells were transfected with bcl-2 via liposome, and the transcription level was evaluated by reverse transcription polymerase chain reaction. The drug resistance was detected by MTT colorimetrical assay. Results：The eucaryotic expression vector pcDNA3.1 （＋）/bcl-2 was constructed successfully. Compared with BIU-87 cells and BIU-87/neo cells transfected with vector pcDNA3.1 （ ＋ ）, the transcription level of bcl-2, the cell survival rate, and the drug resistance were higher in BIU-87 cells transfected with pcDNA3.1 （＋）/bcl-2. Conclusion： The establishinent of MDR cell lines by gene transfection is an ideal method.