摘要
目的研究沙利霉素对葡萄膜恶性黑色素瘤92-1细胞的侵袭和增殖及对β-连环蛋白(β-catenin)/转录因子4(TCF4)表达水平的影响。方法用不同浓度沙利霉素(1,2.5,5,10,20,50,100,150,200μmol·L-1)处理葡萄膜恶性黑色素瘤92-1细胞,48 h后以CCK-8实验检测细胞活力。将92-1细胞随机分为4组:空白对照组和低、中、高3个浓度实验组,空白对照组给予0.9%NaCl,低、中、高浓度实验组分别以终浓度为5,10,20μmol·L-1沙利霉素处理。Transwell实验检测92-1细胞侵袭,克隆形成实验检测92-1细胞增殖,以蛋白质印迹法检测β-catenin和TCF4蛋白表达水平。结果给药48 h后,空白对照组和低、中、高3个浓度实验组侵袭细胞数量分别为76±14,56±12,28±15和10±6;这4组克隆形成率分别为(77±11)%,(54±8)%,(27±9)%和(11±6)%;这4组的β-catenin蛋白相对表达量分别为1.26±0.14,0.57±0.21,0.16±0.08和0.02±0.01;这4组的TCF4蛋白相对表达量分别为0.95±0.12,0.42±0.14,0.12±0.06,0.03±0.02。上述指标:低、中、高3个浓度实验组与空白对照组比较,差异均有统计学意义(均P<0.05)。结论沙利霉素可有效抑制葡萄膜恶性黑色素92-1细胞侵袭及增殖,且具有显著的浓度依赖性。这可能与沙利霉素调节β-catenin/TCF4信号通路有关。
Objective To investigate the effects of salinomycin on invasion and proliferation of uvular malignant melanoma cells 92-1 and expression of β-catenin/transcription factor 4(β-catenin/TCF4). Methods The 92-1 cells of uvular malignant melanoma were treated with different doses of salinomycin(1, 2.5, 5, 10, 20, 50, 100, 150, 200 μmol·L-1), and the cell viability was detected by CCK-8 assay at 48 h after treatment. The 92-1 cells were randomly divided into four groups: blank control group and low, medium and high doses experimental groups. The blank control group was given 0.9% NaCl, and the low, medium and high dose experimental groups were treated with final concentration of 5, 10, 20 μmol·L-1 salinomycin. The 92-1 cell invasion capacity was detected by Transwell assay. The proliferation of 92-1 cells was detected by clone formation assay. The expression of β-catenin and TCF4 proteins were detected by Western blot. Results After 48 h of administration,the number of invasion cells in blank control group,low,medium and high doses experimental groups were 76 ± 14,56 ± 12,28 ± 15 and 10 ± 6,respectively;the clone formation rates in the four groups were(77 ± 11) %,(54 ± 8) %,(27 ± 9) %,and(11 ± 6) %,respectively;the expression of β-catenin protein in the four groups were 1. 26 ± 0. 14,0. 57 ± 0. 21,0. 16 ± 0. 08 and 0. 02 ± 0. 01,respectively;the expression of TCF4 protein in the four groups were 0. 95 ± 0. 12,0. 42 ± 0. 14,0. 12 ± 0. 06 and 0. 03 ± 0. 02. Compared between three doses experimental groups and blank control group,the difference of the factors were significant(all P < 0. 05).Conclusion Salinomycin can effectively inhibit the invasion and proliferation of uvular malignant melanin 92-1 cells in a significant dose-dependent manner. This may be related to the regulation of β-Catenin/TCF4 signaling pathway by salinomycin.
作者
王维
夏玉
邓柏林
高玉英
李静
WANG Wei;XIA Yu;DENG Bo-lin;GAO Yu-ying;LI Jing(Department of Ophthalmology,Sichuan Academy of Medical Sciences·Sichuan Provincial People’s Hospital,Chengdu 610000,Sichuan Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2020年第24期4002-4005,共4页
The Chinese Journal of Clinical Pharmacology
基金
四川省人民医院青年科研基金资助项目(2017QN02)
四川省人民医院眼科-电子科大中央高校基金资助项目(2672018ZYGX2018J098)。
关键词
沙利霉素
葡萄膜恶性黑色素瘤
上皮间质转化
细胞活力
侵袭
增殖
β-连环蛋白/转录因子4信号通路
salinomycin
uveal malignant melanoma
epithelial-mesenchymal transformation
cell viability
invasion
proliferation
β-catenin/transcription factor 4 signaling pathway
