Study on Inhibition Mechanism of Melanogenesis and Melanin Synthesis of B16F10 Cell Induced by Two Natural Products

doi:10.13560/j.cnki.biotech.bull.1985.2017-0122

Abstract

Abstract: This study aims to clarify the inhibition mechanism of proliferation and melanin synthesis of B16F10 cell induced by 10-hydroxycamptothecin（HCPT）and resveratrol（Res）. MTT cytotoxicity assay，microscope observation-assay，L-Dopa oxidation-assay and NaOH cracking-assay were employed to investigate the proliferation，cellular morphology，tyrosinase（TYR）activity and melanin synthesis of B16F10 cells treated by HCPT and Res at different concentrations，respectively；and semi-RT-PCR to analyze the relative mRNA expression of TYR and microphthalmia associated transcription factor（MITF）in B16F10 cells. It was found that HCPT（40，80，120，160，and 200 μmol/L）and Res（80，120，160，and 200 μmol/L）inhibited B16F10 cell proliferation through inducing apoptosis，and exhibited significantly dose-dependent inhibiting effects on tyrosinase activity and melanin synthesis on B16F10 cell（P < 0.05）. In addition，relative mRNA expressions of the TYR and MITF genes in B16F10 cells were depressed remarkably by HCPT at different concentrations and Res at high concentrations（120 and 160 μmol/L）. Conclusively，HCPT and Res may inhibit the B16F10 cell proliferation via inducing cell apoptosis，and depress the melanin synthesis in B16F10 cells from down-regulating the transcription of MITF gene and subsequently inhibiting the gene expression of TYR mRNA and activity of TYR.

Key words: 10-hydroxycamptothecin, resveratrol, tyrosinase, melanin, microphthalmia associated transcription factor

CHENG Xing-an, ZHANG Shu-ming, ZHOU Xiao-wu, WU Bo, LIN Xian-wei, QIN Xiang-jing, HUANG Su-qing, LIU Zhan-mei, JIANG Xu-hong. Study on Inhibition Mechanism of Melanogenesis and Melanin Synthesis of B16F10 Cell Induced by Two Natural Products[J]. Biotechnology Bulletin, 2017, 33(8): 199-205.

References

[1] 袁丹, 容如滨, 喻宗沅等. 抗肿瘤药物10-羟基喜树碱的研究与应用进展[J] . 化学与生物工程, 2007, 12：9-11.
[2] Meng GR, Li JJ, Wang GL, et al. Synthesis and cytotoxic activities of the amino acid-conjugates of 10-hydroxycamptothecin[J] . Chinese Journal of Organic Chemistry, 2014, 1：155-160.
[3] 代鲁平, 宋春霞, 邵先祥, 等. 抗癌药物喜树碱类衍生物的研究进展[J] . 中国药学杂志, 2010, 23：1813-1815.
[4] 王征, 罗泽民, 邓林伟. 白藜芦醇的药理作用机理和合成途径[J] . 天然产物研究与开发, 2003, 15（2）：178-181.
[5] 刘宏胜, 王金环, 徐新女, 等. 白藜芦醇对U251人脑胶质瘤细胞生长的抑制及相关机制的研究[J] . 中国中药杂志, 2009, 34（8）：1027-1031.
[6] 卢晨欣, 孙警辉, 伍春莲. 白藜芦醇与紫杉醇联合用药对人喉癌Hep-2细胞凋亡机制的研究[J] . 中国中药杂志, 2016, 41（3）：476-483.
[7] 吕莎, 李哲, 王显江, 等. 靶向NRP1单克隆抗体对黑色素瘤细胞B16F10的增殖、迁移和凋亡的影响[J] . 中国药学杂志, 2014, 49（20）：1806-1812.
[8] 曾红梅, 张思维, 郑荣寿, 等. 2003~2007年中国皮肤黑色素瘤发病与死亡分析[J] . 中国肿瘤, 2012, 3：183-189.
[9] 唐智柳, 石建伟, 蔡美玉, 等. 中国皮肤黑色素瘤发病及其影响因素分析[J] . 中国肿瘤, 2014, 10：829-833.
[10] 杨娴, 刘汝清, 杜德荣, 等. 白花蛇舌草总黄酮对黑色素瘤a375和b16细胞增殖抑制作用的研究[J] . 世界中西医结合杂志, 2011, 10：844-846, 856.
[11] Vance KW, Goding CR. The transcription network regulating melanocyte development and melanoma[J] . Pigment Cell Research, 2004, 17（4）：318- 325.
[12] Steingrimsson E, Copeland NG, Jenkins NA. Melanocytes and the microphthalmia-transcription factor network[J] . Annual Review of Genetics, 2004, 38：365-411.
[13] Levy C, Khaled M, Fisher DE. M ITF：master regulator of melanocyte development and melanoma oncogene[J] . Trends in Molecular Medicine, 2006, 12：406- 414.
[14] 胡文晋, 胡巍, 方芸. 喜树碱类药物抗肿瘤机制研究进展[J] . 中国药房, 2012, 27：2581-2584.
[15] 张超, 胡巍, 方芸. 雾化吸入羟基喜树碱对小鼠B16黑色素瘤实验性肺转移的影响[J] . 中国中药杂志, 2011, 36（5）：618-623.
[16] Shankar S, Singh G, Srivastava RK. Chemoprevention by resveratrol：molecular mechanisms and therapeutic potential[J] . Frontiers in Bioscience, 2007, 12：4839- 4854.
[17] Joe AK, Liu H, Masumi S, et al. Resveratrol induces growth inhibition, S-phase arrest, apoptosis, and changes in biomarker expression in several human cancer cell lines[J] . Clinical Cancer Research, 2002, 8：893-903.
[18] 陈家顺, 陶方方, 侯敏, 等. 白藜芦醇对鼠纤维肉瘤S180细胞周期及其机制研究[J] . 南京医科大学学报, 2008, 28（5）：570-574.
[19] Bishayee A. Cancer prevention and treatment with resveratrol：from rodent studies to clinical trials[J] . Cancer prevention research（Philadelphia, Pa. ）, 2009, 2（5）：409-418.
[20] 孟德萍, 邹志孟, 许晓群, 等. 白藜芦醇对黑色素瘤细胞增殖的抑制作用[J] . 中华肿瘤防治杂志, 2012, 19（5）：328-331.
[21] Chen YM, Shih TW, Chiu CP, et a1. Effects of lactic acid bacteria-fermented soy milk on mela nogenesis in B16F0 melanocytes[J] . Journal of Functional Foods, 2012, 5：395-405.