华西口腔医学杂志 ›› 2021, Vol. 39 ›› Issue (4): 413-418.doi: 10.7518/hxkq.2021.04.006

• 基础研究 • 上一篇    下一篇

白皮杉醇抗恶性黑色素瘤的体内外实验研究

余波(), 刘伟, 胡敏琪, 唐休发, 李春洁, 阙林()   

  1. 口腔疾病研究国家重点实验室 国家口腔疾病临床医学研究中心 四川大学华西口腔医院头颈肿瘤外科,成都 610041
  • 收稿日期:2020-08-05 修回日期:2021-05-07 出版日期:2021-08-01 发布日期:2021-08-10
  • 通讯作者: 阙林 E-mail:1005137874@qq.com;jy959896@163.com
  • 作者简介:余波,住院医师,硕士,E-mail:1005137874@qq.com
  • 基金资助:
    国家自然科学基金资助项目(81902775);四川大学创新火花库资助项目(2018SCUH0055)

Effect of piceatannol against malignant melanoma in vivo and in vitro

Yu Bo(), Liu Wei, Hu Minqi, Tang Xiufa, Li Chunjie, Que Lin()   

  1. State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Head and Neck Oncology, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2020-08-05 Revised:2021-05-07 Online:2021-08-01 Published:2021-08-10
  • Contact: Que Lin E-mail:1005137874@qq.com;jy959896@163.com
  • Supported by:
    The National Natural Science Foundation of China(81902775);Sichuan University Innovation Spark Project(2018SCUH0055)

摘要: 目的

研究白皮杉醇(PIC)的体内外抗恶性黑色素瘤作用。

方法

体外培养来源于小鼠皮肤的恶性黑色素瘤细胞系B16F10,以梯度浓度PIC处理细胞后甲基噻唑基四唑(MTT)法检测细胞活力;蛋白印迹法检测金属基质蛋白酶(MMP)-2及MMP-9、血管内皮生长因子(VEGF)、脾酪氨酸激酶(Syk)及p-Syk的表达;划痕实验检测迁移能力;Transwell实验检测侵袭能力;再以Syk对应小干扰RNA(si-Syk)沉默Syk进行机制研究。体内实验则以不同浓度PIC对B16F10荷瘤小鼠进行腹腔内注射后完成。

结果

PIC浓度增高使B16F10活力降低;Transwell表明其侵袭能力随PIC浓度增高减弱;划痕实验中PIC浓度增高使愈合时间变长;蛋白印迹表明其主要抑制Syk的磷酸化激活,同时抑制MMP-2、MMP-9、VEGF的表达。经过si-Syk处理能达到对B16F10同样的抑制效果。体内实验则表明PIC可以减慢肿瘤生长。

结论

PIC可以通过抑制Syk的激活抑制恶性黑色素瘤。

关键词: 白皮杉醇, 脾酪氨酸激酶, 恶性黑色素瘤

Abstract: Objective

To study the antitumor effect of piceatannol (PIC) on malignant melanoma in vitro and in vivo.

Methods

B16F10 cells were cultured in vitro and treated with gradient concentrations of PIC. Cell viability was detected with methyl thiazolyl tetrazolium (MTT) assay; matrix metalloproteinase (MMP)-2, MMP-9, vascular endothelial growth factor (VEGF), spleen tyrosine kinase (Syk), and p-Syk were detected with Western blot; migration ability was detected with wound healing assay; invasion ability was detected with Transwell assay. Syk expression was suppressed through RNA interference for the detection of the possible mechanism of PIC in melanoma. An in vivo study was established by creating B16F10-bearing mice with intraperitoneal injection of PIC.

Results

The cell viability of B16F10 decreased with increasing PIC concentration. The results of the Transwell assay showed that invasion ability decreased with increasing PIC concentration, and healing time was prolonged at increased PIC concentration in the wound healing assay. Western blot results showed that PIC mainly inhibited the phosphorylation of Syk and inhibited the expression of MMP-2, MMP-9, and VEGF. RNA interference pointed out that blocking the expression of Syk can reveal the same inhibition effect on B16F10 cells as PIC. In vivo study revealed that different concentrations of PIC cangreatly inhibit melanoma progression.

Conclusion

PIC might block the progression of malignant melanoma by inhibiting spleen tyrosine kinase.

Key words: piceatannol, spleen tyrosine kinase, malignant melanoma

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