CAJ | 학술논문

目的观察全反式维甲酸(ARTA)联合干扰素(IFN)-β对人骨肉瘤细胞株MG-63的生长抑制试验以及对干扰素/维甲酸联合应用诱导凋亡的相关基因19( GRIM-19)和原癌基因信号转导与转录激活因子-3( STAT3)的表达的影响.方法噻唑蓝(MTT)比色法检测不同药物浓度的ATRA/IFN-β,单用或联合应用对MG-63的增殖抑制率；流式细胞仪AnnexinV-FITC/PI法检测MG-63的凋亡；逆转录-聚合酶链反应(RT-PCR)检测MG-63中STAT3与GRIM-19基因的扩增情况；Western blot检测MG-63中STAT3与GRIM-19蛋白的表达.结果 ARTA和IFN-β单用或联合应用均可抑制MG-63的增殖,呈浓度依赖性并与作用时间有关；联合应用抑制作用明显增强,与其他组比较差异有统计学意义(P＜0.05).ARTA、IFN-β单独作用,可诱导MG-63凋亡；联合应用时,诱导凋亡作用显著增强.ARTA联合IFN-β作用,GRIM-19 mRNA的表达明显增强(1.620±0.095),GRIM-19蛋白明显升高(1.850±0.060),与其他组比较差异有统计学意义(P＜0.01).STAT3 mRNA的表达明显降低(0.120 ±0.032),STAT3蛋白明显降低(0.540±0.075),与其他组比较差异有统计学意义(P＜0.01).结论 ARTA/IFN-β作用,可明显抑制MG-63的增殖并诱导其凋亡,其机制可能是诱导GRIM-19高表达,特异性结合于STAT3的转录活性区域,使原癌基因STAT3的表达降低.
목적 관찰전반식유갑산(ARTA)연합간우소(IFN)-β대인골육류세포주MG-63적생장억제시험이급대간우소/유갑산연합응용유도조망적상관기인19( GRIM-19)화원암기인신호전도여전록격활인자-3( STAT3)적표체적영향.방법 새서람(MTT)비색법검측불동약물농도적ATRA/IFN-β,단용혹연합응용대MG-63적증식억제솔；류식세포의AnnexinV-FITC/PI법검측MG-63적조망；역전록-취합매련반응(RT-PCR)검측MG-63중STAT3여GRIM-19기인적확증정황；Western blot검측MG-63중STAT3여GRIM-19단백적표체.결과 ARTA화IFN-β단용혹연합응용균가억제MG-63적증식,정농도의뢰성병여작용시간유관；연합응용억제작용명현증강,여기타조비교차이유통계학의의(P＜0.05).ARTA、IFN-β단독작용,가유도MG-63조망；연합응용시,유도조망작용현저증강.ARTA연합IFN-β작용,GRIM-19 mRNA적표체명현증강(1.620±0.095),GRIM-19단백명현승고(1.850±0.060),여기타조비교차이유통계학의의(P＜0.01).STAT3 mRNA적표체명현강저(0.120 ±0.032),STAT3단백명현강저(0.540±0.075),여기타조비교차이유통계학의의(P＜0.01).결론 ARTA/IFN-β작용,가명현억제MG-63적증식병유도기조망,기궤제가능시유도GRIM-19고표체,특이성결합우STAT3적전록활성구역,사원암기인STAT3적표체강저.
Objective To investigate growth inhibition of human osteosarcoma cell line (MG-63)intervened by all trans retinoic acid (ATRA) combined with interferon (IFN)-β,and the effect on the mRNA expression of gene associated with retinoid-interferon-induced mortality-19 (GRIM-19) and signal transducer and activators of transcription 3 ( STAT3 ).Methods Methyl thiazol tetrazolium (MTT) colorimetric methods were used to test growth inhibition of MG-63 cells intervened by different concentrations of ATRA/IFN-β.Flow cytometry AnnexinV-FITC/PI methods were used to assay apoptosis.Reverse transcription-polymerase chain reaction (RT-PCR) methods were used to detect amplification of GRIM-19 and STAT3 mRNA induced by ATRA combined with IFN-β or alone.Western blotting methods were used to examine the expression of GRIM-19 and STAT3 in MG-63 cells.Results ARTA/IFN-β could inhibit the growth of MG-63 cells in a concentration- and time-dependent manner with the difference being significant in compare with other groups (P ＜0.05).Either ATRA or IFN-β alone could induce apoptosis of MG-63 coulg,but combine use of them could significantly increase apoptosis.The expression of GRIM-19 mRNA ( 1.620 ± 0.095 ) and protein ( 1.850 ± 0.060) was increased significantly in combined group as compared with other groups (P ＜0.01 ).The expression of STAT3 mRNA (0.120 ± 0.032) was decreased significantly in combined group as compared with other groups (P ＜ 0.01 ).Conclusion Combined ARTA wiht IFN-β can obviously inhibit proliferation of MG-63 cells and induce apoptois.The mechanism may be that ARTA/IFN-β induces the high expression of GRIM-19,and the GRIM-19 protein binds with the special domain of STAT3,leading to the decreased expression of STAT3.