西达本胺对恶性黑素瘤细胞系A375的体外抗增殖作用  

In vitro anti-proliferation effect of a histone deacetytase inhibitor, chidamide, on a mall truant melanoma cell line, A375

作　　者：陈佳[1] 周武庆[1] 陈浩[1] 宋亚丽[1] 蔡丽敏[1] 辛崇美[1] 邵雪宝[1] 刘毅[1] 曾学思[1] 孙建方[1] 

机构地区：[1]中国医学科学院北京协和医学院皮肤病研究所病理科,南京210042

出　　处：《中华皮肤科杂志》2009年第4期255-258,共4页Chinese Journal of Dermatology

基　　金：基金项目：江苏省六大高峰人才基金（06-D-072）;江苏省卫生厅科技基金（H200547）

摘　　要：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古抑菌素A对A375细胞体外增殖的影响；AnnexinV—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—500μmol／L浓度范围内呈量效关系，其中50～500μmol／L浓度范围内在作用0～120h时间内呈时效关系。曲古抑菌素A在0．1～1μmol／L浓度范围内呈量效关系，其中在0．25～1μmol／L浓度范围内作用0～120h呈时效关系。西达本胺和曲古抑菌素A作用A375细胞48h的IC50分别为250和0．7μmol／L。西达本胺（62．5，125，250μmol／L）作用48h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（0．175，0．35，0．7μmol／L）作用48h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48h后诱导A375细胞发生GgG。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48h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。Objective To investigate the in vitro anti-proliferation effect of a histune 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 trichostatin 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 MTr 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 - 500 μmol/L and TSA of 0.1 - 1 μmol/L, and in a time-dependent manner from 0 to 120 hours after the beginning of treatment with chidamide of 5 - 500 μmol/L and TSA of 0.25 - 1 μmol/L. The 48-hour 50% growth inhibition concentration （IC50） of chidamide and TSA on A375 cells was about 250 μmol/L and 0.7 μmol/L, respectively. After 48-hour treatment, the apoptosis rate was 80.27% ± 3.06%, 79.53% ± 5.70%, 83.13% ± 6.90% in A375 cells treated with ehidamide 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 untreated 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, 250 μmol/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

关 键 词：肿瘤细胞 培养的 黑色素瘤 西达本胺 曲古抑菌素A 

分 类 号：R739.5[医药卫生—肿瘤] R965[医药卫生—临床医学]