转染抑癌基因KLF6对前列腺癌PC-3细胞的作用研究

Effect of KLF6 on Prostate Cancer Cell Line PC-3 by Transgenic Method

目的:研究抑癌基因KLF6对人前列腺癌细胞系PC-3细胞的生长增殖、细胞周期和对Bc l-2和Cyc lin D1蛋白表达的影响及其可能的作用机制。方法:利用RT-PCR法克隆目的基因KLF6,采用阳离子脂质体介导将含或不含KLF6的pEGFP-C1质粒转染入PC-3细胞,分别作为转染组和对照组。分别进行噻唑蓝(MTT)法观察PC-3细胞的生长抑制率,流式细胞仪检测细胞周期比例变化和凋亡率,免疫组化法观察PC-3细胞Bc l-2和Cyc lin D1的表达水平变化。结果:转染了抑癌基因KLF6的前列腺癌PC-3细胞生长抑制率为(30.0±5.4)%(对照组为0%,P<0.01);细胞周期比例表现为G2/M期减少为(11.2±0.9)%[对照组为(25.2±2.8)%,P<0.05],G0/G1期比例增加为(80.0±9.8)%[对照组为(58.6±7.3)%,P<0.05];细胞凋亡峰为(24.3±2.3)%[对照组为(5.2±0.7)%,P<0.01];Bc l-2的表达率为(18.7±3.2)%[对照组为(41.8±5.9)%,P<0.01];Cyc lin D1的表达率为(25.3±3.7)%[对照组为(38.5±4.6)%,P<0.05]。结论:抑癌基因KLF6的转染可以明显抑制前列腺癌PC-3细胞的生长增殖,并诱导其凋亡,其作用机制可能与下调Bc l-2和Cyc lin D1的表达有关。

Objective： To observe the effect of KLF6 on prostate cancer cell line PC-3 by transgenic method. Methods： We obtained KLF6 cDNA by RT-PCR method from the liver cell, transfected plasmid pEGFP-C1 recombinated with KLF6 into PC-3 cells, and used them as a transfection group and a control group. MTF, flow cytometer and immunocytochemical methods were used to observe the effect of anti-oncogene wild type KLF6 on prostate cancer cell line PC-3 by transgenic method for 48 hours. Results ： After transfected into PC-3 cells, KLF6 enhanced growth suppression, （ 30.0 ± 5.4 ）% in the transfection group and 0% in the control, P〈0.01, apoptosis, （ 24.3 ± 2.3） % in the transfection group and （5.2 ± 0.7 ） % in the control, P〈0.01, the down-regulation of the expression of cyclin D1, （25.3 ± 3.7） % in the transfection group and （ 38.5 ± 4.6） % in the control, P〈0.05 and Bcl-2, （ 18.7 ± 3.2） % in the transfection group, and （41.8 ± 5.9） % in the control, P〈0.01 in PC-3 cells. It also decreased the ratio of the cell phase G2/M, increased the ratio of G0/G1 from （58.6 ± 7.3） % in the control to （ 80.0 ± 9.8 ） % in the transfection group, P〈0.05. Conclu- sion ： PC-3 cells transfected with wild type KLF6 can enhance its growth suppression and apoptosis. It shows great potential for the gene therapy of androgen-independent carcinoma of the prostate.