SOX9基因沉默对膀胱癌EJ细胞株生物学特性的影响及其机制研究  

Effects of SOX9 silencing on biological characteristics of bladder cancer EJ cells and the mechanism

作　　者：楚广民 张建波 孙淼淼 CHU Guangmin;ZHANG Jianbo;SUN Miaomiao(Department of Pathology, Affiliated Tumor Hospital of Zhengzhou University, Zhengzhou 450008, He＇ nan, Chin)

机构地区：[1]郑州大学附属肿瘤医院病理科,郑州4500080

出　　处：《癌症进展》2018年第3期286-289,共4页Oncology Progress

摘　　要：目的探讨SOX9基因沉默对膀胱癌EJ细胞株增殖、凋亡及迁移的影响及其潜在的作用机制。方法采用SOX9 si RNA转染EJ细胞,抑制SOX9基因的表达,并以非特异性si RNA转染EJ细胞为对照。采用Edu细胞荧光染色法检测SOX9基因沉默对EJ细胞增殖的影响,Hoechst 33342细胞染色法检测SOX9基因沉默对EJ细胞凋亡的影响,划痕愈合实验检测SOX9基因沉默对EJ细胞迁移的影响,蛋白质免疫印迹(Western Blot)实验检测PTEN/AKT信号通路相关蛋白的表达水平。结果对照组的SOX9 m RNA表达水平为(100.0±0.5)%,明显高于SOX9沉默组的(14.2±1.3)%,差异有统计学意义(P﹤0.01)。转染72 h后,对照组细胞在450 nm激发光下的OD值为(4.0±0.3),明显高于SOX9沉默组的(2.3±0.2),差异有统计学意义(P﹤0.01)。Edu细胞荧光染色结果显示,转染48 h后,对照组的细胞相对增殖率为(100.0±2.3)%,明显高于SOX9沉默组的(42.2±3.2)%,差异有统计学意义(P﹤0.01)。Hoechst 33342免疫荧光染色结果表明,对照组的细胞凋亡率为(7.2±1.2)%,明显低于SOX9沉默组的(33.2±2.4)%,差异有统计学意义(P﹤0.01)。划痕愈合实验表明,对照组的相对划痕愈合比例为(100.0±1.9)%,明显高于SOX9沉默组的(56.6±4.3)%,差异有统计学意义(P﹤0.01)。SOX9 si RNA转染EJ细胞1 h后,采用Western Blot检测PTEN、AKT及p-AKT蛋白的表达水平。结果显示,SOX9沉默组的PTEN蛋白表达水平高于对照组,p-AKT蛋白表达水平低于对照组。结论沉默SOX9基因能够明显抑制膀胱癌EJ细胞株的增殖和迁移,并促进细胞凋亡,PTEN/AKT信号通路的激活可能与之有关。Objective To explore the effects of SOX9 silencing on the proliferation, apoptosis and migration of EJ cells and the underlying mechanism. Method EJ cells were transfected with SOX9 si RNA to inhibit the expression of SOX9 gene, or with scramble RNA as control. The effects of SOX9 si RNA on cell proliferation were detected by Edu cell immunofluorescence staining assay, cell apoptosis was examined by Hoechst 33342 staining assay, and cell scratch wound healing assay was utilized to determine the effect of SOX9 gene on cell migration, while the expression of PTEN/AKT signaling pathway related protein were evaluated by Western Blot. Result The expression of SOX9 m RNA in control group was（100.0±0.5）%, and was significant higher than that in SOX9 si RNA group at（14.2±1.3）%, the difference was of statistically significance（P〈0.01）. In 72 h after transfection, the OD value at 450 nm exciting light of control group was（4.0±0.3）, which was significantly higher than the value of（2.3±0.2） in SOX9 si RNA group, demonstrating significant difference（P〈0.01）. In Edu cell immunofluorescence staining assay, after 48 h of transfection, the relative rate of proliferation in control group was（100.0±2.3）%, lower than that in SOX9 si RNA group at（42.2±3.2）%, the difference was of statistically significant（P〈0.01）. In Hoechst 33342 staining assay, the apoptosis rate in control group was（7.2 ±1.2）%, which was significantly lower than that in SOX9 si RNA group at（33.2 ± 2.4）%, with significant difference observed（P〈0.01）. In scratch wound healing assay, the relative wound healing rate was（100.0±1.9）% in control group and was significantly higher than that in SOX9 si RNA group at（56.6±4.3）%, showing significant difference（P〈0.01）. The expression of PTEN, AKT and p-AKT protein was evaluated by Western Blot in 1 h after SOX9 si RNA transfection, and the results showed that, PTEN expression increased, while p-AKT decreased in SOX9 si RNA group as compared with c

关 键 词：SIRNA SOX9 膀胱癌 PTEN 

分 类 号：R737.14[医药卫生—肿瘤]