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论文摘要

白皮杉醇抗恶性黑色素瘤的体内外实验研究

Effect of piceatannol against malignant melanoma in vivo and in vitro

作者:余波, 刘伟, 胡敏琪, 唐休发, 李春洁, 阙林

Author:Yu Bo, Liu Wei, Hu Minqi, Tang Xiufa, Li Chunjie, Que Lin

收稿日期:2020-08-05          年卷(期)页码:2021,39(4):413-413-418

期刊名称:华西口腔医学杂志

Journal Name:West China Journal of Stomatology

关键字:白皮杉醇,脾酪氨酸激酶,恶性黑色素瘤,

Key words:piceatannol,spleen tyrosine kinase,malignant melanoma,

基金项目: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)

中文摘要

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。

英文摘要

ObjectiveTo study the antitumor effect of piceatannol (PIC) on malignant melanomain vitroandin vivo.

MethodsB16F10 cells were culturedin vitroand 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. Anin vivostudy was established by creating B16F10-bearing mice with intraperitoneal injection of PIC.

ResultsThe 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 vivostudy revealed that different concentrations of PIC cangreatly inhibit melanoma progression.

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

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