CAJ | 학술논문

目的探讨转化生长因子-β1(TGF-β1)受体拮抗剂LY2109761抑制胃癌细胞侵袭转移的机制.方法利用逆转录-聚合酶链反应(RT-PCR)和Western blot检测TGF-β1及其受体拮抗剂LY2109761作用于胃癌细胞SGC7901后基质金属蛋白酶(MMP)-2、MMP-9、Smad2、血管内皮生长因子(VEGF)等与肿瘤细胞浸润、转移相关基因mRNA及蛋白的表达水平.流式细胞术检测细胞的凋亡率,Transwell迁移侵袭实验检测TGF-β1对细胞迁移及侵袭能力的影响.结果 RT-PCR检测结果显示TGF-β1作用于胃癌细胞时其MMP-2、MMP-9、Smad2、VEGF表达水平分别为0.83 ±0.10、0.74±0.11、0.60±0.08、0.79±0.12,而经LY2109761预处理后分别降为0.45±0.07、0.54±0.14、0.32±0.05、0.51±0.09；Western blot结果显示胃癌细胞经LY2109761预处理后MMP-2、MMP-9、Smad2、VEGF的蛋白表达水平均出现下降趋势；流式细胞术显示经LY2109761预处理后胃癌细胞凋亡率从17.23％下降到5.38％ (P＜0.05)；Transwell迁移及侵袭实验显示胃癌细胞经LY2109761预处理后其迁移及侵袭能力明显减弱(P＜0.05).结论 LY2109761可能是抑制了胃癌细胞的Smad 信号通路、TGF-β诱导的VEGF表达和多种MMPs的活性,从而抑制肿瘤细胞的侵袭及转移能力,具有抗肿瘤作用.
목적 탐토전화생장인자-β1(TGF-β1)수체길항제LY2109761억제위암세포침습전이적궤제.방법 이용역전록-취합매련반응(RT-PCR)화Western blot검측TGF-β1급기수체길항제LY2109761작용우위암세포SGC7901후기질금속단백매(MMP)-2、MMP-9、Smad2、혈관내피생장인자(VEGF)등여종류세포침윤、전이상관기인mRNA급단백적표체수평.류식세포술검측세포적조망솔,Transwell천이침습실험검측TGF-β1대세포천이급침습능력적영향.결과 RT-PCR검측결과현시TGF-β1작용우위암세포시기MMP-2、MMP-9、Smad2、VEGF표체수평분별위0.83 ±0.10、0.74±0.11、0.60±0.08、0.79±0.12,이경LY2109761예처리후분별강위0.45±0.07、0.54±0.14、0.32±0.05、0.51±0.09；Western blot결과현시위암세포경LY2109761예처리후MMP-2、MMP-9、Smad2、VEGF적단백표체수평균출현하강추세；류식세포술현시경LY2109761예처리후위암세포조망솔종17.23％하강도5.38％ (P＜0.05)；Transwell천이급침습실험현시위암세포경LY2109761예처리후기천이급침습능력명현감약(P＜0.05).결론 LY2109761가능시억제료위암세포적Smad 신호통로、TGF-β유도적VEGF표체화다충MMPs적활성,종이억제종류세포적침습급전이능력,구유항종류작용.
Objective To investigate the mechanism of transforming growth factor (TGF)-β1 receptor antagonist LY2109761 inhibiting invasion and metastasis of gastric cancer cells.Methods Human gastric SGC7901 carcinoma cells were treated with TGF-β1 and LY2109761,and the mRNA and protein expression levels of matrix metalloproteinase (MMP)-2,MMP-9,Smad2,vascular endothelial growth factor (VEGF) which were related with invasion and metastasis of tumor cells were analyzed by using reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting.The apoptosis rate was detected by using flow cytometry.Transwell migration and invasion assays were used to detect the migration and invasion abilities of tumor cells before and after the experiment.Results RT-PCR results showed that the expression levels of MMP-2,MMP-9,Smad2 and VEGF were 0.83 ±0.10,0.74 ±0.11,0.60 ±0.08 and 0.79 0.12 respectively in gastric cancer cells treated by TGF-β1,but reduced to 0.45 ± 0.07,0.54 ±0.14,0.32 ±0.05 and 0.51 ±0.09 in gastric cancer cells pretreated by LY2109761.Western blotting revealed that the protein expression levels of MMP-2,MMP-9,Smad2 and VEGF were decreased significantly in gastric cancer cells which were pretreated by LY2109761.The apoptosis rate was decreased from 17.23％ to 5.38％ in gastric cancer cells pretreated by LY2109761 (P ＜ 0.05).The migration and invasion abilities of gastric cancer cells pretreated by LY2109761 were decreased significantly (P ＜ 0.05).Conclusion LY2109761 may inhibit the Smad signaling pathway of gastric cancer cells,TGF-β-induced VEGF expression and a variety of MMPs activity,thereby inhibiting tumor cell invasion and metastasis with the anti-tumor effect.