鸦胆子苦醇联合长波紫外线辐射对人恶性黑色素瘤A375细胞凋亡的影响研究  

Apoptosis effect of brusatol combined with ultraviolet A irradiation on human malignant melanoma A375 cells

作　　者：朱杰 陈君[2] 徐芳 骆文龙 应燕群 ZHU Jie;CHEN Jun;XU Fang;LUO Wen - long;YING Yan- qun(Department of Dermatology ＆ Department of Pharmacy;Department of Dermatology, Hangzhou First People＇s Hospital, Hangzhou 310006, China;Department of Clinical Laboratory, Fuyang Maternal and Child Health Care Hospital, Hangzhou District, Hangzhou 311400, China)

机构地区：[1]杭州市富阳区妇幼保健院皮肤科,杭州311400 [2]杭州市第一人民医院皮肤科,杭州310006 [3]杭州市富阳区妇幼保健院药剂科,杭州311400 [4]杭州市富阳区妇幼保健院检验科,杭州311400

出　　处：《中国临床药理学杂志》2018年第11期1361-1364,共4页The Chinese Journal of Clinical Pharmacology

摘　　要：目的观察鸦胆子苦醇联合长波紫外线(UVA)辐射对人恶性黑色素瘤A375细胞凋亡的影响及机制。方法人恶性黑色素瘤A375细胞分为空白组、对照组和实验组。空白组细胞不做任何处理,对照组用75 k J·m^(-2)长波紫外线处理,实验组在75 k J·m^(-2)长波紫外线处理2 h后分别加入10,20,50,100,200nmol·L^(-1)的鸦胆子苦醇。检测各组细胞活性、细胞周期及凋亡率,检测人转录因子NF-E2相关因子2(Nrf2)通路及线粒体凋亡途径相关蛋白表达情况。结果对照组和实验组细胞活性均显著低于空白组,50,100,200 nmol·L^(-1)实验组细胞活性均显著低于对照组,且随鸦胆子苦醇作用浓度增大而逐渐降低(P<0.05或P<0.01)。对照组和10,20,50,100,200 nmol·L^(-1)实验组细胞凋亡率分别为(3.21±0.24)%,(3.45±0.26)%,(5.86±0.42)%,(12.62±1.12)%,(13.02±2.24)%,(16.15±2.78)%,均显著高于空白组的(0.89±0.12)%,20,50,100,200 nmol·L^(-1)实验组细胞凋亡率均明显高于对照组(均P<0.01)。对照组和实验组与空白组比较,人恶性黑色素瘤A375细胞G0/G1期细胞比例逐渐升高,G2/M和S期细胞比例逐渐降低(P<0.05或P<0.01)。空白组、对照组和50 nmol·L^(-1)实验组Nrf2蛋白相对表达量分别为1.69±0.23,1.23±0.24,1.03±0.15,谷胱甘肽S转移酶P1(GSTP1)蛋白相对表达量分别为2.43±0.16,2.89±0.18,2.78±0.21,B细胞淋巴瘤/白血病-2相关X蛋白(Bax)蛋白相对表达量分别为0.59±0.14,1.05±0.21,1.59±0.24,胱天蛋白酶-3(caspase-3)蛋白相对表达量分别为2.63±0.15,2.71±0.26,2.63±0.28。与空白组比较,对照组和实验组人恶性黑色素瘤A375细胞Nrf2蛋白表达相对量降低,Bax和GSTP1蛋白表达相对量升高,caspase-3无明显变化。结论鸦胆子苦醇联合UVA辐射可抑制人恶性黑色素瘤A375细胞增殖,且存在一定的量效关系,与抑制Nrf2信号通路及激活线粒体凋亡途径相关蛋白,加速细胞凋亡有关。Objective To investigate the effect and mechanism of brusatol combined with ultraviolet A（ UVA） irradiation on apoptosis of human malignant melanoma A375 cells. Methods Human malignant melanoma A375 cells were treated with a dose of 75 k J ·m^（-2） UVA（ control group）, the test group were treated with 10, 20, 50, 100, 200 nmol·L^（-1） brusatol after 75 k J·m^（-2） UVA treating for 2 h,blank group given no treatment. The cell viability,cell cycle and apoptosis rate were measured and the protein expression of transcription factor NF-E2 related factor 2（ Nrf2） pathway and mitochondrial apoptosis pathway were detected. Results The activity of human malignant melanoma A375 cells in control group and test group were lower than that in blank group,the activity of 50,100,200 nmol · L^（-1） test groups were lower than that of control group,the activity of human malignant melanoma A375 cells decreased gradually with the increasing concentration of brusatol（ P〈0. 05 or P〈0. 01）. The cells apoptosis rates of control group and 10,20,50,100,200 nmol·L^（-1） test groups were（ 3. 21 ± 0. 24） %,（ 3. 45 ± 0. 26） %,（ 5. 86 ± 0. 42） %,（ 12. 62 ± 1. 12） %,（ 13. 02 ± 2. 24） %,（ 16. 15 ± 2. 78） %,higher than that of blank group,which was（ 0. 89 ± 0. 12） %,the apoptosis rates of 20,50,100,200 nmol·L^（-1） test groups were significantly higher than that of control group（ all P〈0. 01）. Compared with blank group,the proportion of G0/G1 phase cells in human malignant melanoma A375 cells in control group and 50 nmol·L^（-1） test groups increased gradually and the ratio of G2/M,and S phase cells gradually decreased（ P〈0. 05 or P〈0. 01）. The relative expressions of Nrf2 protein in blank group,control group and test group were 1. 69 ± 0. 23,1. 23 ± 0. 24,1. 03 ± 0. 15, the relative expressions of glutathione S-transferase P1（ GSTP1） protein were2. 43 ± 0. 16,2. 89 ± 0. 18,2. 78 ± 0. 21,the relative expressions of B cell lymphoma/leukemia-2

关 键 词：人恶性黑色素瘤 鸦胆子苦醇 凋亡 

分 类 号：R979.1[医药卫生—药品]