西达本胺联合苦参碱对皮肤T细胞淋巴瘤细胞系增殖、凋亡的影响及可能的凋亡机制  

Effect of chidamide combined with matrine on proliferation and apoptosis of cutaneous T-cell lymphoma cell lines HH and Hut78 and possible apoptotic mechanisms

作　　者：何杏兰 王艺萌[1] 王冠钰 张春雷[1] He Xinglan;Wang yineng;Wang Guanyu;Zhang Chunlei(Department of Dermatology,Peking University Third Hospital,Beijing 100191,China)

机构地区：[1]北京大学第三医院皮肤科,100191

出　　处：《中华皮肤科杂志》2020年第2期121-127,共7页Chinese Journal of Dermatology

基　　金：国家自然科学基金(81372915)。

摘　　要：目的研究西达本胺联合苦参碱对皮肤T细胞淋巴瘤(CTCL)细胞系的增殖抑制和凋亡诱导作用,探讨其凋亡机制。方法采用0.4μmol/L西达本胺、0.6g/L苦参碱单药或联合分别作用HH、Hut78细胞24、48、72 h后,MTS法检测HH、Hut78细胞增殖率。二甲基亚砜(DMSO)处理HH、Hut78细胞作为对照组。西达本胺、苦参碱单药或联合作用两细胞系48 h后,流式细胞仪检测HH和Hut78细胞凋亡率,Western免疫印迹法检测各组细胞凋亡相关蛋白的表达。统计分析采用重复测量、单因素方差分析,组间两两比较采用LSDV检验。结果与对照组相比,西达本胺、苦参碱单药或联合均能一定程度抑制HH、Hut78细胞增殖(F=15.88.558.26,P<0.05、<0.001)048 h时,Hut78细胞系苦参碱组(20.98%±1.53%)、西达本胺组(22.44%±7.74%)和联合组细胞凋亡率(44.53%±1.85%)均显著高于对照组(8.42%±4.23%;LSD-t=4.76,5.31、13.69,均P<0.05),且联合组显著高于苦参碱组和西达本胺组(LSD-t=8.93、8.37,均P<0.01)。HH细胞系苦参碱组(13.98%±3.86%)、西达本胺组细胞凋亡率(13.61%±1.62%)与对照组(11.44%±1.43%)相比差异无统计学意义(均P>0.05),但联合组(20.94%±0.64%)显著高于对照组、苦参碱组和西达本胺组(LSD-t=7.37,5.40,5.69,均P<0.05)。HH细胞系中,联合组caspase-3剪切体蛋白表达显著高于对照组、苦参碱组和西达本胺组(均P<0.05),而E-cadherin、p65、p-Bad及Bcl-2蛋白表达显著低于其他3组(均P<0.05)。Hut78细胞系中,苦参碱组、西达本胺组和联合组E-cadherin、p65>p-Bad、Bcl-2蛋白表达均显著低于对照组(均P<0.05),仅西达本胺组和联合组caspase-3剪切体蛋白表达显著高于对照组(均P<0.05)。HH及Hut78细胞4组Bad蛋白表达差异均无统计学意义(均P>0.05)。结论西达本胺联合苦参碱可抑制HH、Hut78细胞增殖,诱导其凋亡,其机制可能与调控细胞凋亡相关蛋白E-cadherin、p65、p-Bad、Bcl-2及caspase-3剪切体蛋白表达有关。Objective To evaluate the effect of chidamide combined with matrine on proliferation and apoptosis of cutaneous T-cell lymphoma(CTCL)cell lines HH and Hut78,and to explore their apoptotic mechanisms.Methods Both HH and Hut78 cells were treated with 0.4 μmol/L chidamide and 0.6 g/L matrine alone or in combination for 24,48 and 72 hours,with those treated with dimethyl sulfoxide(DMSO)serving as control groups.MTS assay was performed to detect cellular proliferation rates of HH and Hut78 cells at each time point.After 48-hour treatment,flow cytometry was conducted to detect cell apoptosis,and Western blot analysis to determine expression of apoptosis-related proteins in these cells.Statistical analysis was carried out by using repeated measures analysis of variance,one-way analysis of variance,and least significant difference(LSD)-t test for multiple comparisons.Results Compared with DMSO,chidamide and matrine alone or in combination could inhibit the proliferation of HH and Hut78 cells to different extents(F=15.88,558.26,P<0.05,<0.001,respectively).At 48 hours,the apoptosis rate in Hut78 cells was significantly higher in the matrine group(20.98%±1.53%),chidamide group(22.44%±7.74%)and combination group(44.53%±1.85%)than in the control group(8.42%±4.23%;LSD-t=4.76,5.31,13.69 respectively,all P<0.05),as well as in the combination group than in the matrine group and chidamide group(LSD-t=8.93,8.37 respectively,both P<0.01);no significant differences were observed in the apoptosis rate of HH cells between the matrine group(13.98%±3.86%)or chidamide group(13.61%±1.62%)and control group(11.44%±1.43%,both P>0.05),while the combination group(20.94%±0.64%)showed a significantly higher apoptosis rate compared with the control group,matrine group and chidamide group(LSD-Z=7.37,5.40,5.69 respectively,all P<0.05).In the case of HH cells,the combination group showed significantly higher cleaved caspase-3 expression(all P<0.05),but significantly lower protein expression of E-cadherin,nuclear factor(NF)-κB,phosphorylated

关 键 词：淋巴瘤 T细胞 皮肤 苦参碱 细胞凋亡 西达本胺 Hut78细胞 HH细胞 

分 类 号：R73[医药卫生—肿瘤]