雷公藤红素对人恶性黑色素瘤细胞凋亡的影响  

Effects of tripterine on inducing apoptosis of human malignant melanoma cells

作　　者：饶俊珍 汪娟[1] Rao Junzhen;Wang Juan(Department of Pharmacy,Wuhan First Hospital,Wuhan Hospital of Integrated Traditional Chinese and Western Medicine,Hubei Province,Wuhan 432000,China)

机构地区：[1]湖北省武汉市第一医院武汉市中西医结合医院药学部,武汉432000

出　　处：《中国医药》2022年第8期1242-1246,共5页China Medicine

摘　　要：目的观察雷公藤红素对人恶性黑色素瘤MUM-2C细胞凋亡的影响,并探究其可能机制。方法体外培养黑色素瘤MUM-2C细胞,采用不同浓度(0、0.125、0.25、0.5、1、2、4、8、16、32μmol/L)的雷公藤红素处理黑色素瘤MUM-2C细胞,采用细胞计数试剂盒8(CCK-8)检测细胞活性确定实验浓度,根据选定浓度将黑色素瘤MUM-2C细胞分为对照组和低、中、高浓度组及抑制剂组;对照组无特殊处理,低、中、高浓度组分别采用1、2、4μmol/L的雷公藤红素处理,抑制剂组采用蛋白激酶R样内质网激酶(PERK)抑制剂GSK2656157处理。采用流式细胞仪检测各组细胞周期及凋亡情况;采用蛋白质印迹法检测各组凋亡蛋白B淋巴细胞瘤2(Bcl-2)和Bcl相关X蛋白(Bax)表达情况;采用蛋白质印迹法检测各组细胞PERK信号通路蛋白及内质网应激标志蛋白免疫球蛋白重链结合蛋白(Bip)的表达情况。结果 根据CCK-8实验结果确定培养24、48、72 h的半抑制浓度分别为2.14、2.03、1.98μmol/L,设置实验作用浓度为低剂量组1μmol/L,中剂量组2μmol/L,高剂量组4μmol/L。使用低、中、高浓度雷公藤红素处理组和抑制剂组G/G期细胞比例、细胞的凋亡率[(15.2±3.0)%、(26.3±4.2)%、(37.3±5.0)%、(40.5±5.3)%比(8.4±2.1)%]、Bax蛋白相对表达水平明显高于对照组(均P<0.05),且对雷公藤红素呈剂量依赖性。使用雷公藤红素处理各组和抑制剂组G/M期和S期细胞比例、Bcl-2、磷酸化PERK和Bip蛋白相对表达水平明显低于对照组(均P<0.05),且对雷公藤红素呈剂量依赖性。结论雷公藤红素可以促进黑色素瘤MUM-2C细胞的凋亡,其作用机制可能与抑制PERK通路的激活介导细胞内质网应激,增加凋亡蛋白的表达诱导细胞凋亡相关。Objective To observe the effects of tripterine on the apoptosis of human malignant melanoma MUM-2 C cells, and to explore its possible mechanism. Methods The melanoma MUM-2 C cells were cultured in vitro and were treated with tripterine of different concentrations(0, 0.125, 0.25, 0.5, 1, 2, 4, 8, 16, 32 μmol/L). The cell activity was detected by cell counting kit-8(CCK-8) to determine the experimental concentration. According to the determined concentration, melanoma MUM-2 C cells were divided into control group, low concentration group, medium concentration group, high concentration group and inhibitor group. The control group had no special treatment;the low, medium and high concentration groups were treated with 1, 2 and 4 μmol/L tripterine, respectively;the inhibitor group was treated with protein kinase R-like ER kinase(PERK) inhibitor GSK2656157. The cell cycle and apoptosis in each group were detected by flow cytometry. The expressions of apoptosis proteins B lymphocytoma 2(Bcl-2), Bcl-associated X protein(Bax), PERK signaling pathway proteins and endoplasmic reticulum stress marker immunoglobulin heary chain binding protein(Bip) in each group were detected by western blotting. Results According to CCK-8 results, the 50% inhibiting concentration concentrations for 24, 48 and 72 h were 2.14, 2.03 and 1.98 μmol/L, respectively. The experimental concentrations were set as 1 μmol/L in low dose group, 2 μmol/L in medium dose group, and 4 μmol/L in high dose group. Compared with control group, proportion of cells in G/Gphase, apoptosis rate[(15.2±3.0)%,(26.3±4.2)%,(37.3±5.0)%,(40.5±5.3)% vs(8.4±2.1)%] and the relative expression level of Bax protein were significantly increased in low, medium and high concentration tripterine group and inhibitor group(all P<0.05), showing dose-dependence on tripterine. Compared with control group, proportion of cells in G/M phase and S phase, and the relative expression levels of Bcl-2, phosphorylation PERK and Bip proteins were significantly decreased in each group t

关 键 词：恶性黑色素瘤细胞 雷公藤红素 细胞凋亡 凋亡蛋白 蛋白激酶R样内质网激酶抑制剂 

分 类 号：R739.5[医药卫生—肿瘤]