两种天然产物对B16F10细胞增殖及黑色素合成抑制机理研究

发布时间：2018-02-12 18:51

本文关键词： -羟基喜树碱白藜芦醇酪氨酸酶黑色素小眼相关转录因子　出处：《生物技术通报》2017年08期 　论文类型：期刊论文

【摘要】：旨在研究10-羟基喜树碱(10-hydroxycamptothecin,HCPT)和白藜芦醇(Resveratrol,Res)对体外培养的小鼠恶性黑色素瘤B16F10细胞的增殖及黑色素合成抑制机理。利用MTT法、显微观察、L-Dopa氧化法、Na OH裂解法分析不同浓度HCPT和Res对细胞增殖、细胞形态、酪氨酸酶活性及黑色素合成含量的影响。荧光半定量PCR方法(Semi-RT-PCR)分析该化合物对黑色素合成关键因子酪氨酸酶(TYR)和小眼相关转录因子(MITF)基因表达的影响。结果表明HCPT(40、80、120、160和200μmol/L)和Res(80、120、160和200μmol/L)能够通过诱导细胞凋亡抑制B16F10细胞的增殖,同时对酪氨酸酶活性和细胞黑色素生成具有明显抑制作用(P0.05),且呈现浓度依赖性。另外,不同浓度的HCPT以及高浓度Res(120和160μmol/L)能够显著下调B16F10细胞TYR和MITF基因的mRNA水平。HCPT和Res可能通过诱导细胞凋亡抑制B16F10细胞的增殖,同时通过下调MITF基因转录,抑制TYR m RNA的表达及TYR酶活性,进而抑制细胞黑色素的生成。
[Abstract]:The aim of this study was to study the inhibitory mechanism of 10-hydroxycamptothecin HCPTand resveratrol on the proliferation and melanin synthesis of murine malignant melanoma B16F10 cells cultured in vitro. The cell proliferation and morphology of different concentrations of HCPT and Res were analyzed by L-Dopa oxidation method. The effects of tyrosinase activity and melanin synthesis on the expression of tyrosinase tyrosinase tyrosinase and MITFs in melanin synthesis were analyzed by semi-quantitative PCR semi-quantitative PCR method. The results showed that the expression of tyrosinase tyrosinase and transcriptional factor MITFs were related to melanin synthesis. HCPTT (40, 80, 80, 120, 160 and 200 渭 mol / L) and Resch80 (120, 200 渭 mol / L) could inhibit the proliferation of B16F10 cells by inducing apoptosis. At the same time, the activity of tyrosinase and the formation of cytomelanin were significantly inhibited by P0.05, and in a concentration-dependent manner. Different concentrations of HCPT and high concentrations of Res(120 and 160 渭 mol / L could significantly down-regulate the mRNA level of TYR and MITF genes in B16F10 cells. HCPT and Res could inhibit the proliferation of B16F10 cells by inducing apoptosis, and down-regulate the transcription of MITF gene. It inhibited the expression of TYR m RNA and the activity of TYR, and then inhibited the production of cytomelanin.
【作者单位】：仲恺农业工程学院天然产物化学研究所;中国科学院南海海洋研究所;
【基金】：国家自然科学基金项目(21406274,21376281) 广东省大学生创新训练项目(201411347034,201511347056)
【分类号】：R96

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