CAJ | 학술논문

目的观察肝癌缺失基因-1(DLC-1)和黏着斑激酶(FAK)对肿瘤OVCAR-3细胞增殖和凋亡的影响.方法构建DLC-1表达载体、FAK特异性小发卡RNA(shRNA)载体和FAK-shRNA联合DLC-1表达载体,分别转染OVCAR-3细胞,应用实时定量聚合酶链反应(Real-time PCR)和Western blot检测OVCAR-3细胞中DLC-1和p-FAK(Y397)的表达,细胞计数试剂盒(CCK-8)和流式细胞仪检测细胞增殖和凋亡,Western blot检测磷酸化细胞外信号调节激酶1/2 (p-ERK1/2)的表达.结果 DLC-1表达增加或/和FAK基因沉默后,OVCAR-3细胞增殖显著受抑(0.737±0.094,P＜0.05)、凋亡明显增加(15.21±1.05)％ (P＜0.05),p-ERK1/2表达水平显著降低(0.151±0.010,P＜0.05).结论 DLC1可能通过调节FAK及其下游的ERK信号通路参与肿瘤细胞的增殖和凋亡.
목적 관찰간암결실기인-1(DLC-1)화점착반격매(FAK)대종류OVCAR-3세포증식화조망적영향.방법 구건DLC-1표체재체、FAK특이성소발잡RNA(shRNA)재체화FAK-shRNA연합DLC-1표체재체,분별전염OVCAR-3세포,응용실시정량취합매련반응(Real-time PCR)화Western blot검측OVCAR-3세포중DLC-1화p-FAK(Y397)적표체,세포계수시제합(CCK-8)화류식세포의검측세포증식화조망,Western blot검측린산화세포외신호조절격매1/2 (p-ERK1/2)적표체.결과 DLC-1표체증가혹/화FAK기인침묵후,OVCAR-3세포증식현저수억(0.737±0.094,P＜0.05)、조망명현증가(15.21±1.05)％ (P＜0.05),p-ERK1/2표체수평현저강저(0.151±0.010,P＜0.05).결론 DLC1가능통과조절FAK급기하유적ERK신호통로삼여종류세포적증식화조망.
Objective To investigate the effects of deleted in liver cancer-1 (DLC-1) and focal adhesion kinase (FAK) on the growth and apoptosis of malignant OVCAR-3 cells.Methods DLC-1 expression plasmid,FAK specific small hairpin RNA (shRNA) expression plasmid,and FAK-shRNA combined DLC-1 expression plasmid were constructed and transfected into OVCAR-3 cells.Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting were used to detect the expression of DLC-1 and FAK in OVCAR-3 cells,cell proliferation and apoptosis were examined by cell counting kit-8 (CCK-8) kit and flow cytometry,and the expression of phospho-extracellular signal-regulated kinase 1/2 (p-ERK1/2) was measured by using Western blotting.Results While the DLC-1 expression increased or/and FAK knocked down,the growth of OVCAR-3 cells was obviously inhibited [(0.737 ± 0.094),P ＜0.05],but apoptosis was significantly induced [(15.21 ± 1.05) ％,P ＜ 0.05].In addition,the expression of p-ERK1/2 was notably decreased [(0.151 ±0.010),P ＜0.05].Conclusion DLC1 might be implicated in the growth and apoptosis of malignant tumor cells via its regulation on FAK and downstream ERK signaling.