[1]杨历辉,李东霞,苏依拉,等.小檗碱对人皮肤黑素瘤A375细胞周期相关miRNA的影响[J].中国皮肤性病学杂志,2016,(11):1123-1126,1134.[doi:10.13735/j.cjdv.1001-7089.201605119]
 YANG Li-hui,LI Dong-xia,SU Yi-la,et al.Inhibitory Effect of Berberine on Human Melanoma A375 Cells[J].The Chinese Journal of Dermatovenereology,2016,(11):1123-1126,1134.[doi:10.13735/j.cjdv.1001-7089.201605119]
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小檗碱对人皮肤黑素瘤A375细胞周期相关miRNA的影响
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《中国皮肤性病学杂志》[ISSN:1001-7089/CN:61-1197/R]

卷:
期数:
2016年11期
页码:
1123-1126,1134
栏目:
论著
出版日期:
2016-11-01

文章信息/Info

Title:
Inhibitory Effect of Berberine on Human Melanoma A375 Cells
作者:
杨历辉1李东霞1苏依拉2苏秀兰2
1.内蒙古医科大学附属医院,内蒙古 呼和浩特 010050; 2.内蒙古医科大学附属医院临床医学研究中心,内蒙古 呼和浩特 010050
Author(s):
YANG Li-hui1LI Dong-xia1SU Yi-la2SU Xiu-lan2
1.Affiliated Hospital of Inner Mongolia Medical University, Hohehot, Inner Mongolia 010050, China; 2.Clinical Medical Research Center of the Affiliated Hospital, Inner Mongolia Medical University,No.1 Tongdao North Street,Hohehot, Inner Mongolia 010050, China
关键词:
小檗碱 黑素瘤 miRNA 细胞增殖 细胞周期
Keywords:
Berberine Melanoma miRNA Cell proliferation Cell cycle
分类号:
R739.5
DOI:
10.13735/j.cjdv.1001-7089.201605119
文献标志码:
A
摘要:
目的 探讨小檗碱对皮肤黑素瘤A375细胞株增殖及细胞周期相关miRNA及其靶基因表达的影响。 方法 体外培养A375细胞,加入不同浓度小檗碱 (0,20,40,60,80,100μmol/L),用噻唑蓝(MTT)法测定小檗碱对A375细胞生长的抑制作用; 流式细胞实验检测细胞周期; 实时定量RT-PCR法检测小檗碱对A375细 胞miRNA-34a,miRNA-154,miRNA -26a,miRNA -124及其对应靶基因CDK4,CyclinD1,CyclinE,CDK2表达的影响。采用SPSS13.0软件进行多因素方差分析。结果 MTT 法结果显示小檗碱抑制A375细胞增殖,具有时效依赖性和量效依赖性(P<0.05)。流式细胞实验表明低浓度(20,40μmol/L)小檗碱作用后细胞周期呈S 期阻滞和G2/M期阻滞(均P<0.05),高浓度(60,80μmol/L)小檗碱作用后细胞周期呈G2/M期阻滞(P<0.05)。实时定 量RT-PCR结果表明,小檗碱作用后随着浓度的增加miRNA-34a,miRNA-154,miRNA -26a,miRNA -124表达增加(均P<0.05),其对应的靶基因 CDK4,CyclinD1,CyclinE,CDK2表达减弱(均P<0.05)。结论 小檗碱对A375细胞的生长抑制作用呈时间和浓度依赖性; 小檗碱可能通过调控细胞周期相 关miRNA及其靶基因对黑素瘤A375细胞发挥抑制作用。
Abstract:
Objective To investigate the effects of berberine on the proliferation and cell cycle related miRNA and and its target gene expression in cutaneous melanoma A375 cell line.Methods A375 cells were cultured in vitro, and clssified into various groups to be treated with berberine at different concentrations(0,20,40,60,80,100μM).Subsequently,methyl thiazolyl tetrazolium(MTT)assay was performed to evaluate cell growth. Cell cycle was detected by flow cytometry(FCM). QRT-PCR was conducted to measure the miRANs of miRNA-34a, miRNA-154, miRNA-26a, miRNA-124 and their corresponding genes of CDK4, Cyclin D1, Cyclin E,CDK2.Statistical analysis was done by multi-way analysis of variance(ANOVA)using the software SPSS13.0.Results MTT assay suggested berberine can inhibit A375cells proliferation in a dose and time dependent manner(P<0.05).Berberine at low doses(20,40μmol/L)promoted S and G2/M arrest,and higher doses(60μmol/L,80 μmol/L)resulted in G2/M arrest by FCM.Compared with the control group, QRT-PCR results showed that the effect of berberine,with concentration increase,the expressions of miRNA-34a, miRNA-154, miRNA -26a and miRNA-124 were increased, and the expressions of their corresponding genes CDK4,Cyclin D1,Cyclin E,CDK2 were decreased(both P<0.05).Conclusion Berberine inhibits the growth of A375 cells in a dose-and time-dependent manner,and may induce the inhibition via affecting key factors of cell cycle.

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备注/Memo

备注/Memo:
[基金项目] 2014年内蒙古自治区科技计划项目(kjt14sf16); 2016年内蒙古自治区自然科学基金面上项目(2016MS08137) [通讯作者] 李东霞,E-mail: ldx828@sina.com
更新日期/Last Update: 2016-11-01