CAJ | 학술논문

目的研究组蛋白去乙酰化酶抑制剂西达本胺对体外培养的皮肤恶性黑素瘤细胞株A375细胞的抗癌作用.方法用不同浓度的西达本胺和曲古抑菌素A处理A375细胞,以噻唑蓝法检测西达本胺和曲占抑菌素A对A375细胞体外增殖的影响;Annexin V-EGFP/PI双荧光活染-流式细胞测量法检测细胞凋亡率;DNA倍体分析检测细胞周期.结果西达本胺和曲古抑菌素A可明显抑制A375细胞增殖,西达本胺在5～500 μmol/L浓度范围内呈量效关系,其中50-500 μmol/L浓度范围内在作用0～120 h时间内呈时效关系.曲古抑菌素A在0.1～1 μmol/L浓度范围内呈量效关系,其中在0.25～1 μmol/L浓度范围内作用0～120 h呈时效关系.西达本胺和曲古抑菌素A作用A375细胞48 h的IC50分别为250和0.7 μmol/L.西达本胺(62.5,125,250 μmol/L)作用48 h后诱导A375细胞的凋亡率分别为80.27%±3.06%、79.53%±5.70%、83.13%±6.90%,曲古抑菌素A(0.175,0.35,0.7 μmol/L)作用48 h后诱导A375细胞的凋亡率分别为16.27%±2.46%、28.83%±2.55%、83.40%±8.65%,显著高于空白对照组(10.43%±0.96%)(P＜0.01).西达本胺(62.5,125,250μmol/L)作用48 h后诱导A375细胞发生G0/G1期阻滞,G0/G1期细胞比例分别为76.30%±6.06%、82.79%±0.74%、88.91%±5.29%,与空白对照组(38.73%±3.36%)比较,差异有统计学意义(P＜0.01);曲古抑菌素A(0.35,0.7μmol/L)作用48 h后诱导A375细胞发生G2/M期阻滞,G2/M期细胞比例分别为25.15%±2.71%、58.71%±3.45%,与空白对照组(15.73%±0.23%)比较,差异有统计学意义(P＜0.01).结论西达本胺和曲古抑菌素A在体外能诱导A375细胞发生周期阻滞,促使细胞凋亡,抑制细胞生长.
목적 연구조단백거을선화매억제제서체본알대체외배양적피부악성흑소류세포주A375세포적항암작용.방법 용불동농도적서체본알화곡고억균소A처리A375세포,이새서람법검측서체본알화곡점억균소A대A375세포체외증식적영향;Annexin V-EGFP/PI쌍형광활염-류식세포측량법검측세포조망솔;DNA배체분석검측세포주기.결과 서체본알화곡고억균소A가명현억제A375세포증식,서체본알재5～500 μmol/L농도범위내정량효관계,기중50-500 μmol/L농도범위내재작용0～120 h시간내정시효관계.곡고억균소A재0.1～1 μmol/L농도범위내정량효관계,기중재0.25～1 μmol/L농도범위내작용0～120 h정시효관계.서체본알화곡고억균소A작용A375세포48 h적IC50분별위250화0.7 μmol/L.서체본알(62.5,125,250 μmol/L)작용48 h후유도A375세포적조망솔분별위80.27%±3.06%、79.53%±5.70%、83.13%±6.90%,곡고억균소A(0.175,0.35,0.7 μmol/L)작용48 h후유도A375세포적조망솔분별위16.27%±2.46%、28.83%±2.55%、83.40%±8.65%,현저고우공백대조조(10.43%±0.96%)(P＜0.01).서체본알(62.5,125,250μmol/L)작용48 h후유도A375세포발생G0/G1기조체,G0/G1기세포비례분별위76.30%±6.06%、82.79%±0.74%、88.91%±5.29%,여공백대조조(38.73%±3.36%)비교,차이유통계학의의(P＜0.01);곡고억균소A(0.35,0.7μmol/L)작용48 h후유도A375세포발생G2/M기조체,G2/M기세포비례분별위25.15%±2.71%、58.71%±3.45%,여공백대조조(15.73%±0.23%)비교,차이유통계학의의(P＜0.01).결론 서체본알화곡고억균소A재체외능유도A375세포발생주기조체,촉사세포조망,억제세포생장.
Objective To investigate the in vitro anti-proliferation effect of a histone deacetylase inhibitor,chidamide,on a cutaneous malignant melanoma cell line,A375.Methods Cultured A375 cells were treated with different concentrations of chidamide(5,10,50,100,500 μmol/L)and aichostatin A (TSA)(0.1,0.25,0.5,1.0 μmol/L),respectively,for various durations(24,48,72,96,120 hours).Subsequently,cell proliferation,apoptosis and cell cycle were detected by MTT assay,annexin Vfluorescein isothiocyanate and propidium iodide double staining,and DNA ploid analysis,respectively.Results The proliferation of A375 cells was inhibited in a dose-dependent manner by chidamide of 5-500μmol/L and TSA of 0.1-1 μmol/L,and in a time-dependent manner from 0 to 120 hours after the beginning of trealment with ehidamide of 5-500μmol/L and TSA of 0.25-1μmol/L.The 48-hour 50% growth inhibition concentration(IC50)of ehidamide and TSA on A375 cells was about 250 μmol/L and 0.7μmol/L,respectively.After 48-hour treatment,the apoptosis mte was 80.27%±3.06%,79.53%±5.70%,83.13%±6.90%in A375 cells treated with chidamide of 62.5,125,250 μmol/L,respectively,16.27%±2.46%,28.83%±2.55%,83.40%±8.65%in those treated with TSA of 0.175,0.35,0.7 μmol/L,respectively,10.43%±0.96%in ontreated cells;a statistical increase was noticed in chidamide-treated cells and TSA-treated cells vs.untreated cells(all P＜0.001).A positive correlation was observed between the apoptosis rate and concentrations of TSA(r=0.955,P=0.000).Cell cycle analysis indicated that treatment with chidamide induced cell cycle arrest in G0/G1 phase,with the cell proportion in G0/G1 phase being 76.30%±6.06%,82.79%±0.74%,88.91%±5.29%in A375 cells treated with chidamide of 62.5,125,250μmol/L,respectively,versus 38.73%±3.36%in untreated cells.While after 48-hour treatment with TSA of 0.35 and 0.7 μmol/L,the proportion of cells in G2/M phases was 25.15%±2.71%and 58.71%±3.45%,respectively,compared to 15.73%±0.23%in untreated cells(P＜0.01).Conclusion Chidamide and TSA could induce cell cycle arrest and apoptosis,as well as inhibit the growth of A375 ceils in vitro.