iNOS基因转染对雄激素非依赖性前列腺癌细胞DU145生长的影响

Transfection of iNOS suppresses the growth of androgen-independent prostate cancer DU145 cells

目的:观察诱导型一氧化氮合酶(iNOS)基因转染对雄激素非依赖性前列腺癌细胞DU145生物学行为的影响。方法:将iNOS基因转染到DU145细胞并筛选出阳性细胞进行扩增,并设空载体组和对照组。观察细胞的形态变化,MTT法绘制生长曲线;流式细胞术检测细胞凋亡率;了解NOS抑制剂对转染细胞的影响。结果:转染iNOS后,DU145细胞分泌的NO[(272.50±15.82)μmol/L]显著高于空载体组[(122.00±18.93)μmol/L]和对照组[(121.00±6.98)μmol/L](P<0.05)。流式细胞术检测结果提示转染iNOS组细胞凋亡率[(42.78±2.01)%]明显高于空载体组[(30.65±1.46)%]和对照组[(28.96±1.50)%](P<0.05)。MTT测定结果提示转染组细胞生长较空载体组和对照组减慢(P<0.05),NOS抑制剂可以加快其生长,但无显著性差异(P>0.05)。结论:iNOS基因转染可以使DU145细胞分泌较高浓度的NO,诱导细胞凋亡,抑制细胞生长,为晚期雄激素非依赖性前列腺癌的基因治疗提供一个有效的靶点。

[ Abstract ] Objective： To observe the effect of the inducible nitric oxide synthase （iNOS） gene on androgen-independent pros- tate cancer DU145 cells in vitro. Methods： The iNOS gene was transfeeted into androgen-independent prostate cancer DU145 cells. The positive cells were selected as the transfected group for amplification, and an empty vector （EV） group and a control group were also set. The mRNA transcription of iNOS was analyzed by RT-PCR. The morphological changes of the cells were observed, the effect of iNOS transfection on the cell growth determined using the MTr method, and the apoptosis of DU145 cells detected by flow eytometry, followed by analysis of the effect of NOS inhibitors on the transfected cells. Results ： DU145 cells transfected with iNOS secreted significantly more nitric oxide （ [272.50 ±15.82] μmol/L） than those of the EV and control groups （ [ 122.00 ±18.93 ] μmol/L and [ 121.00 ±6.981 μmol/L） （P 〈0.05）. The rate of cell apoptosis was markedly enhanced in the transfected group as compared with the EV and control groups （ [42.78 ±2.01 1% vs [30.65 ± 1.46]% and [28.96 ± 1.50]%, P 〈0.05）. MTT test indicated a slower growth of the DU145 cells in the former than in the latter two （P 〈0.05）. NOS inhibitors enhanced their growth, but with no significance （P 〉 0.05）. Conclusion： DU145 cells transfected with iNOS could secrete high-concentration nitric oxide, induce cell apoptosis, and suppress cell proliferation, which may provide a potentially effective gene therapy for advanced androgen-independent prostate cancer.