中华皮肤科杂志 ›› 2018, Vol. 51 ›› Issue (6): 456-459.doi: 10.3760/cma.j.issn.0412-4030.2018.06.016

• 研究报道 • 上一篇    下一篇

小檗碱抑制人黑素瘤A375细胞增殖的机制研究

任敏1,李东霞2,杨凌3,苏秀兰3   

  1. 1. 内蒙古医科大学附属医院皮肤性病科
    2. 内蒙古医科大学附属医院皮肤科
    3. 内蒙古医科大学附属医院临床医学研究中心
  • 收稿日期:2017-03-27 修回日期:2017-12-22 发布日期:2018-05-30
  • 通讯作者: 李东霞 E-mail:ldx828@sina.com
  • 基金资助:
    内蒙古自治区科技计划项目;内蒙古自治区自然科学基金

A study on mechanism underlying the inhibition of berberine on proliferation of human melanoma A375 cells

Ren-Min 1,Dong-Xia LI 3, 3   

  • Received:2017-03-27 Revised:2017-12-22 Published:2018-05-30
  • Contact: Dong-Xia LI E-mail:ldx828@sina.com
  • Supported by:
    Science and Technology Planning Project of Inner Mongolia Autonomous Region;Natural Science Foundation of Inner Mongolia Autonomous Region of China

摘要: 目的 探讨小檗碱对人黑素瘤A375细胞周期相关miRNA及其靶基因表达的影响。方法 体外培养人黑素瘤A375细胞,经不同浓小檗碱(20、40、60、80 μmol/L)处理48 h后,采用qRT-PCR检测miRNA-582-5p及其靶基因CDK1 mRNA的表达,以及miRNA-188-5p和其靶基因CDK2、Cyclin D1、Cyclin A mRNA的表达。Western印迹检测细胞周期相关蛋白CDK1、CDK2、Cyclin D1和Cyclin A的表达。结果 qRT-PCR显示,与空白对照组相比,20、40 μmol/L小檗碱组miRNA-582-5p的表达无明显变化(均P > 0.05),60、80 μmol/L组miRNA-582-5p的表达明显上调(均P < 0.05)。与空白对照组相比,各浓度小檗碱组miRNA-188-5p表达均增强(F = 22.600,P = 0.002),CDK2、CDK1、Cyclin A、Cyclin D1mRNA表达随小檗碱浓度增加逐渐减弱(F = 51.976、248.510、626.671、312.740,均P < 0.001)。Western印迹显示,小檗碱可降低CDK1、CDK2、Cyclin D1、Cyclin A蛋白的表达(F = 138.124、110.966、278.772、140.167,均P < 0.001)。结论 小檗碱可能通过降低mRNA表达量与升高miRNA-582-5p和miRNA-188-5p含量共同使黑素瘤A375细胞周期相关蛋白CDK1、CDK2、Cyclin D1和Cyclin A表达降低,进而阻滞细胞周期进程,抑制肿瘤生长。

关键词: 小檗碱, 黑色素瘤, 微RNAs, 细胞周期蛋白类, 细胞周期蛋白依赖激酶类, A375细胞

Abstract: Ren Min, Li Dongxia, Yang Ling, Su Xiulan Department of Dermatology and Venereology, Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010050, China (Ren M, Li DX); Clinical Medical Research Center, Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010050, China (Yang L, Su XL) Corresponding author: Li Dongxia Email: ldx828@sina.com 【Abstract】 Objective To evaluate the effect of berberine on the of cell cycle-related miRNA and its target gene in human melanoma A375 cells. Methods In vitro cultured A375 cells were classified into several groups to be treated with berberine at different concentrations of 0 (blank control group), 20, 40, 60, 80, and 100 μmol/L, respectively, for 48 hours. qRT-PCR was performed to determine the mRNA of miRNA-582-5p and its target gene cyclin-dependent kinase 1 (CDK1), miRNA-188-5p and its target gene CDK2, Cyclin D1 and Cyclin A. Western blot analysis was conducted to measure the protein of cell cycle-related proteins CDK1, CDK2, Cyclin D1 and Cyclin A. Results qRT-PCR showed that compared with the blank control group, the 20 and 40 μmol/L berberine groups had a similar of miRNA-582-5p (both P > 0.05), but the 60 and 80 μmol/L berberine groups had a significantly up-regulated of miRNA-582-5p (both P < 0.05). Compared with the blank control group, all the 5 berberine groups had a significantly increased of miRNA-188-5p (F = 22.600, P = 0.002). However, the mRNA of CDK2, CDK1, Cyclin A and Cyclin D1 gradually decreased along with the increase of berberine concentrations (F = 51.976, 248.510, 626.671 and 312.740, respectively, all P < 0.001). Western blot analysis revealed that berberine decreased the protein of CDK1, CDK2, Cyclin D1 and Cyclin A (F = 138.124, 110.966, 278.772 and 140.167, all P < 0.001). Conclusion Berberine can decrease the of cell cycle-related proteins CDK1, CDK2, Cyclin D1 and Cyclin A, likely by decreasing their mRNA and increasing the of miRNA-582-5p and miRNA-188-5p, and then block the cell cycle progression of A375 cells and inhibit the growth of tumor.

Key words: Berberine, Melanoma, MicroRNAs, Cyclins, Cyclin-dependent kinases, A375 cells

引用本文

任敏 李东霞 杨凌 苏秀兰. 小檗碱抑制人黑素瘤A375细胞增殖的机制研究[J]. 中华皮肤科杂志, 2018,51(6):456-459. doi:10.3760/cma.j.issn.0412-4030.2018.06.016

Ren-Min Dong-Xia LI. A study on mechanism underlying the inhibition of berberine on proliferation of human melanoma A375 cells[J]. Chinese Journal of Dermatology, 2018, 51(6): 456-459.doi:10.3760/cma.j.issn.0412-4030.2018.06.016