CAJ | 학술논문

目的:体外实验研究三氧化二砷(As2O3)对人胃癌阿霉素耐药细胞株(SGC7901/ADM)的逆转耐药机制.方法:MTT法检测磷酸化ERK(p-ERK)激动剂G-CSF作用前后As2O3的逆转耐药倍数;免疫细胞化学法测定As2O3及G-CSF作用前后SGC7901/ADM细胞内Ras和p-ERK的变化;流式细胞仪测定G-CSF和As2O3干预后SGC7901/ADM的细胞周期和凋亡率.结果:SGC7901/ADM对ADM耐药,As2O3可逆转耐药,其中0.5μmol/L As2O3作用48 h后逆转耐药倍数约为6.29.G-CSF干预后,逆转耐药倍数降至4.72;SGC7901/ADM中Ras表达高于亲本细胞株SGC7901/S,而p-ERK无明显差异,As2O3可下调Ras及p-ERK的表达.G-CSF干预后,As2O3下调Ras及p-ERK表达的能力较干预前显著降低.0.1μmol/L和0.5μmol/L As2O3组的G0~G1期细胞比例和凋亡率均显著高于各个对照组;G-CSF干预后同一剂量的As2O3组G0~G1期细胞及凋亡率较干预前均显著降低.结论:As2O3可逆转人胃癌耐药细胞株SGC7901/ADM对ADM的耐药作用,其机制与下调Ras/p-ERK信号传导通路中Ras、磷酸化ERK的表达有关.
목적:체외실험연구삼양화이신(As2O3)대인위암아매소내약세포주(SGC7901/ADM)적역전내약궤제.방법:MTT법검측린산화ERK(p-ERK)격동제G-CSF작용전후As2O3적역전내약배수;면역세포화학법측정As2O3급G-CSF작용전후SGC7901/ADM세포내Ras화p-ERK적변화;류식세포의측정G-CSF화As2O3간예후SGC7901/ADM적세포주기화조망솔.결과:SGC7901/ADM대ADM내약,As2O3가역전내약,기중0.5μmol/L As2O3작용48 h후역전내약배수약위6.29.G-CSF간예후,역전내약배수강지4.72;SGC7901/ADM중Ras표체고우친본세포주SGC7901/S,이p-ERK무명현차이,As2O3가하조Ras급p-ERK적표체.G-CSF간예후,As2O3하조Ras급p-ERK표체적능력교간예전현저강저.0.1μmol/L화0.5μmol/L As2O3조적G0~G1기세포비례화조망솔균현저고우각개대조조;G-CSF간예후동일제량적As2O3조G0~G1기세포급조망솔교간예전균현저강저.결론:As2O3가역전인위암내약세포주SGC7901/ADM대ADM적내약작용,기궤제여하조Ras/p-ERK신호전도통로중Ras、린산화ERK적표체유관.
Objective:The aim of this in vitro study is to assess the reversing effects of arsenic trioxide (As2O3) on multi-drug-resistance (MDR) of SGC7901/adriamycin (ADM) and determine the molecular mechanism associated with Ras/p-ERK downregulation in human gastric cancer cells. Methods:The reversing effects of As2O3 before and after the treatment with an ERK agonist were determined by methyl thiazolyl tetrazolium assay. The molecular mechanism involved was further studied by examining the expressions of Ras and phosphorylated p-ERK by immunocytochemical assay. The process was repeated in pre-and post-intervention of As2O3 and granulocyte colony stimulating factor (G-CSF), respectively. Changes in the cell cycle and apoptotic rates were determined by flow cytometry. Results:SGC7901/ADM was resistant to ADM, and As2O3 could reverse the drug resistance. After the treatment of As2O3 at a dose of 0.5 μmol/L for 48 h, the the multiple of reversed drug resistance was about 6.29 approximately, which was greater compared with that of the normal cells. After administration of As2O3, the IC50 decreased dramatically, which shows that As2O3 could reverse MDR. After the G-CSF intervention, the multiple of the reversing effects was only 4.72. Immunocytochemical assay showed higher expressions of Ras in SGC7901/ADM than in the parental cell line SGC7901/S. No obvious difference in the p-ERK expressions of these cell lines was observed. As2O3 could decrease both Ras and p-ERK expressions (q-test, P<0.01). After pre-treatment with the ERK agonist, Ras and p-ERK expressions decreased (t-test, P<0.01). Flow cytometry indicated that the proportion of G0-G1 cells and apoptosis rate were higher in the As2O3–treated group (0.1 and 0.5μmol/L) than in the control groups. And apoptotic rate of both the cells significantly decreased in the As2O3 group of the same dose after treatment of G-CSF. Conclusion:As2O3 can reduce the MDR of gastric cancer in vitro via the Ras/p-ERK signaling transduction pathway.