番茄红素对人皮肤鳞状细胞癌COLO16细胞关键信号受体调控研究  

Regulatory effects of lycopene on the key signaling receptors in human cutaneous squamous cell carcinoma cell line COLO16

作　　者：毕素云 李莉[2] 徐松[2] 张孟丽[2] 顾恒[2] 周之海[1] 陈旭[2] Bi Suyun;Li Li;Xu Song;Zhang Mengli;Gu Heng;Zhou Zhihai;Chen Xu(Department of Dermatology and Venereology, Tianjin Medical University General Hospital, Tianjin 300070, Chin;Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs, Nanfing 210042, China（)

机构地区：[1]天津医科大学总医院皮肤性病科,300070 [2]中国医学科学院北京协和医学院皮肤病研究所江苏省皮肤病与性病分子生物学重点实验室

出　　处：《中华皮肤科杂志》2018年第6期421-424,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81773342、81673083）;教育部博士点基金（20131106120046）;重大疾病创新研究（2016x320014）;中国医学科学院医学与健康科技创新工程项目（2017-12M-1-017）

摘　　要：目的探讨番茄红素对人皮肤鳞状细胞癌细胞系COL016细胞关键信号受体调控的影响。方法COL016细胞分为0（对照组）、5、10、15、20、25μmol／L番茄红素6个处理组，处理24h后进行LDH实验确定对细胞活力的影响。根据LDH分泌量确定细胞死亡水平，选取番茄红素对细胞作用的安全浓度信号受体表达量实验，蛋白免疫印迹检测关键信号受体蛋白表达水平。使用SPSS软件进行单因素方差分析，采用Tukey Multiple Comparison和Brown-Forsythe（B）检验方法。结果5、10、15、20、25μmol／L番茄红素处理COL016细胞24h后，25μmol／L组细胞死亡率和对照组比差异有统计学意义（F=13．116，P〈0．05），选取5、10、20μmol／L三个浓度番茄红素处理COL016细胞24h后，表皮生长因子受体蛋白磷酸化水平降低（P〈0．05），糖皮质激素受体蛋白表达水平升高（P〈0．05），而维A酸受体、雄激素受体、孕激素受体蛋白表达水平变化不显著。5、10、20μmol／L番茄红素处理HaCaT、HEK细胞24h后，HaCaT细胞和人原代角质形成细胞中表皮生长因子受体蛋白磷酸化水平和糖皮质激素受体蛋白表达量差异无统计学意义（均P〉0．05）。结论番茄红素可降低COL016细胞活力，抑制表皮生长因子受体蛋白活化，上调糖皮质激素受体表达，这种效应可能有肿瘤细胞特异性。Objective To evaluate the regulatory effects of lycopene on the key signaling receptors in human cutaneous squamous cell carcinoma cell line COLO16. Methods Cultured COL016 cells were divided into 6 groups to be treated with lycopene at different concentrations of 0, 5, 10, 15, 20, and 25 μ mol/L, respectively, for 24 hours （control group and 5, 10, 15, 20, 25μmol/L lycopene groups）, followed by estimation of the cell viability by lactate debydrogenase （LDH） assay. Lycopene at a safe concentration was selected based on the LDH assay, and used for the determination of expression of signaling receptors, and Western blot analysis was performed to measure the expression of key signaling receptor proteins, including epidermal growth factor receptor （EGFR）, glucocorticoid receptor （GR）, retinoic acid receptor-alpha （RAR-α）, retinoid X receptor-alpha （RXR-α）, androgen receptor （AR） and progesterone receptor （PR）. Statistical analysis was carried out by one-way analysis of variance （ANOVA）, Tukey multiple comparison teat and Brown-Forsythe test with the SPSS software. Results After 24-hour treatment with lycopene at different concentrations, there were significant differences in the rate of cell death among these groups （F = 13.116, P 〈 0.05）, and the rate of cell death in the 25 μmol/L lycopene group significantly differed from that in thecontrol group （P 〈 0.05）. Therefor, lycopene at concentrations of 5, 10 and 20 μmol/L were selected to treat COLO16 and HaCaT cells as well as human epidermal keratinocyte （HEK） for 24 hours in the following experiment. The treatment with lycopene significantly decreased the phosphorylation level of EGFR （P 〈 0.05）, but significantly increased the expression of GR protein （P 〈 0.05）, and showed no significant effects on the protein expression of RAR-α, RXR-α, AR, and PR in COLO16 cells. After 24-hour treatment with lycopene at concentrations of 5, 10 and 20 μmol/L, there were no significant changes in the phosphor

关 键 词：癌 鳞状细胞 细胞系 肿瘤 受体 表皮生长因子 受体 糖皮质激素 COL016细胞 番茄红素 

分 类 号：R739.5[医药卫生—肿瘤]