天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (8): 818-823.doi: 10.11958/20202793

• 实验研究 • 上一篇    下一篇

组蛋白去乙酰化酶抑制剂与PD98059缀合物对皮肤鳞状细胞癌的疗效研究

张良，陈娜，付昱，郑亮，肖婧，陈柳青   

  1. 武汉市第一医院皮肤科（邮编430000）
  • 收稿日期:2020-10-12 修回日期:2021-03-24 出版日期:2021-08-15 发布日期:2021-08-19
  • 通讯作者: 张良 E-mail:xiaokela2009@126.com
  • 作者简介:张良（1977），男，硕士，副主任医师，主要从事皮肤外科方面研究。E-mail：xiaokela2009@126.com
  • 基金资助:
    湖北省卫生计生科研项目（WJ2019M028）

The therapeutic effect of histone deacetylase inhibitor and PD98059 conjugate on cutaneous squamous cell carcinoma

ZHANG Liang, CHEN Na, FU Yu, ZHENG Liang, XIAO Jing, CHEN Liu-qing   

  1. Department of Dermatology, Wuhan No.1 Hospital, Wuhan 430000, China
  • Received:2020-10-12 Revised:2021-03-24 Published:2021-08-15 Online:2021-08-19

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摘要： 目的 探讨组蛋白去乙酰化酶（HDAC）抑制剂与PD98059缀合物的体内、外抗皮肤鳞状细胞癌（SCC）作 用。方法 采用HDAC试剂盒测试缀合物SAHA-PD98059对HDAC活性的抑制作用；采用5-［3-（羧基甲氧基）苯 基］-3-（4，5-二甲基-2-噻唑基）-2-（4-磺基苯基）-2H-四唑内盐（MTS）试剂盒测试缀合物SAHA-PD98059对A431、 HSC-5、Colo16 和 SCC-12 4 种 SCC 细胞增殖的抑制作用。构建 A431 皮下荷瘤鼠模型，并将其随机分为 SAHAPD98059组、SAHA组、PD98059组和空白对照组，连续灌胃给药18 d，每隔2 d记录小鼠体质量，测量肿瘤体积。通过 对小鼠主要器官行HE染色评价缀合物SAHA-PD98059的生物安全性。结果 缀合物SAHA-PD98059抑制HDAC 活性的半抑制浓度（IC50）为（142.9±1.2）nmol/L。缀合物 SAHA-PD98059 在 HDAC 活性口袋中能够与氨基酸残基 H142、G151、T312 形成氢键，而母体化合物 SAHA 可与氨基酸残基 H142、H143、T312 形成氢键。与母体化合物 PD98059及SAHA相比，缀合物SAHA-PD98059对皮肤鳞癌细胞A431、HSC-5、Colo16和SCC-12的抗增殖活性均更 强，其中缀合物SAHA-PD98059对A431细胞增殖的抑制作用最强［IC50=（1.7±0.2）µmol/L］。缀合物SAHA-PD98059 对人正常皮肤细胞TE353.sk基本无毒性。体内研究表明，缀合物SAHA-PD98059的半数致死量（LD50）为647.5 mg/kg， 并可显著抑制 A431 肿瘤的增长。HE 染色发现，缀合物 SAHA-PD98059 对主要器官无明显毒性。结论 缀合物 SAHA-PD98059用于治疗SCC安全有效，其效果强于单一使用SAHA或PD98059。

关键词: 皮肤肿瘤, 癌, 鳞状细胞, 组蛋白脱乙酰基酶类, PD98059, 伏立诺他, 缀合物, 抗肿瘤活性

Abstract: Objective To investigate the effect of histone deacetylase (HDAC) inhibitor and PD98059 conjugate on squamous cell carcinomas (SCC) activity in vitro and in vivo. Methods HDAC inhibitory activity of conjugate SAHAPD98059 was evaluated by HDAC assay kit. The antiproliferative activity of conjugate SAHA-PD98059 on A431, HSC-5, Colo16 and SCC-12 was assessed by 5-[3-(carboxymethoxy)phenyl]-3-(4,5-dimethyl-2-thiazolyl)-2-(4-sulfophenyl)-2Htetrazolium inner salt (MTS) assay kit. A431 subcutaneous tumor-bearing mouse model was constructed by continuous intragastric administration for 18 days. The model mice were randomly divided into conjugate SAHA-PD98059 group, SAHA group, PD98059 group and blank control group. The body weight and tumor size of mice were recorded every 2 days. The biological safety of conjugate SAHA-PD98059 was evaluated by H&E staining of main organs of mice. Results The half inhibitory concentration (IC50) of conjugate SAHA-PD98059 inhibitory HDAC activity was (142.9±1.2) nmol/L. Conjugate SAHA-PD98059 was able to form hydrogen bonds with amino acid residues H142, G151 and T312 in HDAC active pocket, and SAHA engaged in hydrogen bonds with amino acid residues H142, H143 and T312. Compared with the parent compound PD98059 and SAHA, the conjugate SAHA-PD98059 displayed stronger anti-proliferative activity against A431, HSC-5, Colo16 and SCC-12. Among them, conjugate SAHA-PD98059 showed the greatest potency against A431 (IC50=1.7 mmol/L±0.2 mmol/L). Moreover, conjugate SAHA-PD98059 was nontoxic to health TE353. sk cells. In vivo biological evaluation exhibited that median lethal dose (LD50) of conjugate SAHA-PD98059 was 647.5 mg/kg, and which could significantly inhibit the growth of A431 tumor in mice. Meanwhile, HE histology data showed that there were no noticeable toxic effects of conjugate SAHA-PD98059 on the major organs. Conclusion The conjugate SAHA-PD98059 is safe and effective for the treatment of SCC, and it its effect is stronger than that the single use of SAHA or PD98059

Key words: skin neoplasms, carcinoma, squamous cell, histone deacetylases, PD98059, SAHA, conjugate, anti-tumor activity

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