CAJ | 학술논문

目的抑制Glioma-associated oncogene homoglog （ GLI）基因在卵巢癌SKOV3细胞中的表达，探讨GLI1表达抑制对SKOV3细胞增殖的影响。方法构建并筛选GLI1基因的RNAi表达质粒，转染SKOV3细胞，CCK-8法比较转染前后卵巢癌细胞增殖变化，流式细胞仪检测紫杉醇诱导转染前后细胞凋亡比例，流式细胞仪检测转染前后SKOV3的细胞周期情况。 Western blot 检测cyclinD、Bcl-2及Capase3蛋白表达的影响。结果转染48 h后，在SKOV3细胞中Control组、N-Control组、GLI1 shRNA-1组、GLI1 shRNA-2组、GLI1 shRNA-3组、GLI1 shRNA-4组的GLI1 mRNA水平分别为1.00±0.00、1.03±0.02、0.73±0.07、0.98±0.08、0.53±0.08、0.68±0.04，与Control组相比，GLI1 shRNA-1、GLI1 shRNA-3、GLI1 shRNA-4组GLI1 mRNA表达量均下降（ P＜0.05），转染GLI1 shRNA-3的SKOV3细胞中GLI1蛋白表达水平低于GLI1 shRNA-1、GLI1 shRNA-2、GLI1 shRNA-4组（ P＜0.05），具有显著的干扰效果。转染GLI1 shRNA-3的SKOV3细胞增殖受到明显抑制（ P＜0.05），在紫杉醇的诱导下细胞凋亡比例明显增加，细胞周期主要阻滞在G1/S期。 Western blot检测发现Bcl-2蛋白表达下降（P＜0.05），Capase3蛋白表达升高（P＜0.05），细胞周期蛋白CyclinD1蛋白表达下降（P＜0.05）。结论 GLI1 shRNA对SKOV3细胞增殖抑制作用明显，促进细胞凋亡，GLI1信号可通过抑制cyclinD1活化抑制细胞恶性增殖。
목적억제Glioma-associated oncogene homoglog （ GLI）기인재란소암SKOV3세포중적표체，탐토GLI1표체억제대SKOV3세포증식적영향。방법구건병사선GLI1기인적RNAi표체질립，전염SKOV3세포，CCK-8법비교전염전후란소암세포증식변화，류식세포의검측자삼순유도전염전후세포조망비례，류식세포의검측전염전후SKOV3적세포주기정황。 Western blot 검측cyclinD、Bcl-2급Capase3단백표체적영향。결과전염48 h후，재SKOV3세포중Control조、N-Control조、GLI1 shRNA-1조、GLI1 shRNA-2조、GLI1 shRNA-3조、GLI1 shRNA-4조적GLI1 mRNA수평분별위1.00±0.00、1.03±0.02、0.73±0.07、0.98±0.08、0.53±0.08、0.68±0.04，여Control조상비，GLI1 shRNA-1、GLI1 shRNA-3、GLI1 shRNA-4조GLI1 mRNA표체량균하강（ P＜0.05），전염GLI1 shRNA-3적SKOV3세포중GLI1단백표체수평저우GLI1 shRNA-1、GLI1 shRNA-2、GLI1 shRNA-4조（ P＜0.05），구유현저적간우효과。전염GLI1 shRNA-3적SKOV3세포증식수도명현억제（ P＜0.05），재자삼순적유도하세포조망비례명현증가，세포주기주요조체재G1/S기。 Western blot검측발현Bcl-2단백표체하강（P＜0.05），Capase3단백표체승고（P＜0.05），세포주기단백CyclinD1단백표체하강（P＜0.05）。결론 GLI1 shRNA대SKOV3세포증식억제작용명현，촉진세포조망，GLI1신호가통과억제cyclinD1활화억제세포악성증식。
Objective To explore the effects of GLI 1 signaling on the proliferation of ovarian cancer cell . Methods Ovarian cancer SKOV3 cells were transfected for 48 hours by the mediation of Lipofectamine TM 2000. CCK-8 kit and real time RT-PCR were employed to study the effects of GLI 1 mRNA transfection on the proliferation of SKOV3 cell,the apoptosis rate induced by paclitaxel detected by flow cytometry ,the cell cycle of SKOV3 cells following the treatments was measured with flow cytometry and the expression of cyclinD 1 protein detected by western blot.Results In the Control,negative-control group,GLI1 shRNA-1 group,GLI1 shRNA-2 group,GLI1 shRNA-3 group,GLI1 shRNA-4 group,of SKOV3 cells,the expression of GLI1 mRNA relative to GAPDH was 1.00 ±0.00, 1.03 ±0.02,0.73 ±0.07,0.98 ±0.08,0.53 ±0.08,0.68 ±0.04,respectively.Contrast to Control group,the GLI1 mRNA expression in GLI1 shRNA-1 group,GLI1 shRNA-3 group,GLI1 shRNA-4 group declined (P <0.05), moreover the GLI1 mRNA expression in GLI1 shRNA-3 group obviously lower than other 3 groups ( P <0.05 ) . Meanwhile the expression of GLI 1 protein decreased .SKOV3 cell proliferation and CyclinD 1 protein significantly inhibited under the condition of transfected by GLI 1 shRNA-3.The apoptosis rate increased induced by paclitaxel after GLI1 down-regulated.The expression of Bcl-2 and CyclinD1 were decreased and the expression of Caspase 3 was increased.Conclusions GLI1 shRNA visibly inhibit the proliferation of ovarian cancer cell and induce the cell apoptosis,GLI1 signaling could induce cell proliferation by actived CyclinD 1.