茶多酚对小鼠黑色素瘤细胞B16F10体外增殖和迁移的影响

doi:10.16431/j.cnki.1671-7236.2022.04.009

摘要/Abstract

摘要： 【目的】研究茶多酚对小鼠黑色素瘤细胞B16F10增殖和迁移的影响,以期为将茶多酚用于肿瘤的治疗提供依据。【方法】用不同浓度茶多酚(0.05、0.10和0.15 g/L)处理B16F10细胞,不处理(0 g/L)的作为对照组,采用CCK-8法检测各组细胞生长情况,计算生长抑制率;不同浓度茶多酚处理细胞24 h后,采用划痕试验评价细胞的迁移能力;通过实时荧光定量PCR和Western blotting检测B16F10细胞中小眼畸形相关转录因子(microphthalmia-associated transcription factor,MITF)和黑色素形成限速酶(tyrosinase,TYR)mRNA和蛋白的表达水平。【结果】与对照组相比,0.05 g/L茶多酚对B16F10细胞增殖无明显抑制作用(P>0.05),0.10和0.15 g/L茶多酚对B16F10细胞的增殖能力有极显著的抑制作用(P<0.01),且浓度越高,抑制作用越强;0.05、0.10和0.15 g/L茶多酚对B16F10细胞的迁移均有极显著的抑制作用(P<0.01);实时荧光定量PCR和Western blotting结果显示,随着茶多酚浓度的升高,MITF和TYR mRNA和蛋白的表达量均呈浓度依赖性降低,与对照组相比,0.05 g/L茶多酚显著降低MITF mRNA和蛋白的表达(P<0.05),0.10和0.15 g/L茶多酚均极显著降低MITF mRNA和蛋白的表达(P<0.01),0.10和0.15 g/L茶多酚均极显著降低TYR mRNA和蛋白的表达(P<0.01)。【结论】 0.10和0.15 g/L茶多酚对B16F10细胞的增殖和迁移均有明显的抑制作用,并且能降低黑色素生成基因MITF和TYR的表达,表明茶多酚能抑制黑色素瘤的进展。

关键词: 黑色素瘤B16F10细胞; 茶多酚; 增殖; 迁移

Abstract: 【Objective】 This study was aimed to study the effects of tea polyphenols on the proliferation and migration of mouse melanoma cell B16F10,in order to provide evidence for the use of tea polyphenols in tumor therapy.【Method】 B16F10 cells were treated with different concentrations (0.05,0.10 and 0.15 g/L) of tea polyphenols,no treatment (0 g/L) was used as the control group.CCK-8 was used to determine the cell growth of different treatment groups,and the growth inhibition rate was calculated.After treated with different concentrations of tea polyphenols for 24 h,the migration ability of the cells was evaluated by scratch test.The expression of microphthalmia-associated transcription factor(MITF) and tyrosinase(TYR) mRNA and protein in B16F10 cells were detected by Real-time quantitative PCR and Western blotting.【Result】 Compared with control group,0.05 g/L tea polyphenols had no significant inhibitory effect on the proliferation of B16F10 cells (P>0.05),0.10 and 0.15 g/L tea polyphenols had extremely significant inhibitory effects on B16F10 cells (P<0.01),and the higher the concentration,the stronger the inhibition.0.05,0.10 and 0.15 g/L tea polyphenols had extremely significant inhibitory effects on the migration of B16F10 cells (P<0.01).Real-time quantitative PCR and Western blotting results showed that as the concentration of tea polyphenols increased,the expressions of MITF and TYR mRNA and protein gradually decreased in a concentration-dependent manner.Compared with control group,0.05 g/L tea polyphenols could significantly reduce the expressions of MITF mRNA and protein (P<0.05),0.10 and 0.15 g/L tea polyphenols could extremely significantly decrease the expression of MITF mRNA and protein (P<0.01).0.10 and 0.15 g/L tea polyphenols could extremely significantly decrease the expressions of TYR mRNA and protein (P<0.01).【Conclusion】 0.10 and 0.15 g/L tea polyphenols had significant inhibitory effect on the proliferation and migration of B16F10 cells,and could reduce the expression of melanogenesis genes MITF and TYR,indicating that tea polyphenols could inhibit the progression of melanoma.

Key words: melanoma B16F10 cells; tea polyphenols; proliferation; migration

中图分类号:

R285

刘芳君, 蓝吴涛, 李鹏飞, 许冬梅, 张磊, 朱芷葳. 茶多酚对小鼠黑色素瘤细胞B16F10体外增殖和迁移的影响[J]. 中国畜牧兽医, 2022, 49(4): 1280-1286.

LIU Fangjun, LAN Wutao, LI Pengfei, XU Dongmei, ZHANG Lei, ZHU Zhiwei. Effect of Tea Polyphenols on Proliferation and Migration of Mouse Melanoma Cell B16F10 in vitro[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1280-1286.

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