[1]唐剑辉,黄昭明,余盼盼,等.知母皂苷AⅢ调控miR-494对原发性皮肤黑色素瘤生物学行为的影响[J].陕西中医,2022,(10):1354-1358.[doi:DOI:10.3969/j.issn.1000-7369.2022.10.007]
　TANG Jianhui,HUANG Zhaoming,YU Panpan,et al.Effects of timosaponin AⅢ on biological behavior of primary cutaneous melanoma by regulating miR-494[J].,2022,(10):1354-1358.[doi:DOI:10.3969/j.issn.1000-7369.2022.10.007]
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知母皂苷AⅢ调控miR-494对原发性皮肤黑色素瘤生物学行为的影响
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《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2022年10期
页码:
1354-1358
栏目:
基础研究
出版日期:
2022-10-05

文章信息/Info

Title:
Effects of timosaponin AⅢ on biological behavior of primary cutaneous melanoma by regulating miR-494
作者:
唐剑辉; 黄昭明; 余盼盼; 朱必胜; 刘启胜
(咸宁市中心医院,湖北 咸宁 437100)
Author(s):
TANG Jianhui; HUANG Zhaoming; YU Panpan; ZHU Bisheng; LIU Qisheng
(Xianning Central Hospital,Xianning 437100,China)
关键词:
知母皂苷AⅢ;  miR-494;  黑色素瘤;  增殖;  迁移和侵袭
Keywords:
Timosaponin AⅢ;  miR-494;  Melanoma;  Poliferation;  Migration and invasion
分类号:
R 739.5
DOI:
DOI:10.3969/j.issn.1000-7369.2022.10.007
文献标志码:
A
摘要:
目的:探讨知母皂苷AⅢ调控miR-494对原发性皮肤黑色素瘤增殖、迁移和侵袭的影响。方法:体外培养人黑色素瘤A375细胞,分别用0、4、8、16 μmol/L的知母皂苷AⅢ处理A375细胞24 h。将miR-NC、miR-494转染至A375细胞,作为miR-NC、miR-494组; 将anti-miR-NC、anti-miR-494转染至A375细胞,再用知母皂苷AⅢ(16 μmol/L)干预,作为anti-miR-NC+16 μmol/L组、anti-miR-494+16 μmol/L组。细胞计数试剂盒-8(CCK-8)检测细胞增殖情况; 蛋白质印迹(Western blot)法检测细胞Cyclin D1、MMP-2、MMP-9蛋白表达; Transwell实验检测细胞迁移和侵袭情况; 实时荧光定量 PCR(RT-qPCR)检测细胞miR-494表达水平。结果:与对照组比较,8、16 μmol/L中、高剂量组的知母皂苷AⅢ明显下调组细胞A值、迁移数量和侵袭数量以及Cyclin D1、MMP-2、MMP-9蛋白表达(P<0.05)。与对照组比较,8 μmol/L中剂量组、16 μmol/L高剂量组的知母皂苷AⅢ明显上调细胞miR-494表达水平(P<0.05)。与miR-NC组比较,miR-494组细胞A值、迁移数量和侵袭数量以及Cyclin D1、MMP-2、MMP-9蛋白表达明显降低(P<0.05)。与anti-miR-NC+16 μmol/L组比较,anti-miR-494+16 μmol/L组细胞miR-494表达水平显著降低; 细胞A值、迁移数量和侵袭数量以及Cyclin D1、MMP-2、MMP-9蛋白表达显著升高(P<0.05)。结论:知母皂苷AⅢ上调miR-494抑制原发性皮肤黑色素瘤增殖、迁移和侵袭。
Abstract:
Objective:To investigate the effects of miR-494 regulated by Anemarrhena asphodeloides saponin AⅢ on proliferation,migration and invasion of primary cutaneous melanoma.Methods:Human melanoma A375 cells were cultured in vitro and treated with 0,4,8 and 16 μmol/L Anemarrhena asphodeloides saponin AⅢ for 24 h.miR-NC and miR-494 were transfected into A375 cells as miR-NC and miR-494 groups; anti-miR-NC and anti-miR-494 were transfected into A375 cells,and then treated with Anemarrhena asphodeloides saponin AⅢ(16 μmol/L)as anti-miR-NC +16 μmol/L group and anti-miR-494 +16 μmol/L group.Cell counting kit-8(CCK-8)was used to detect cell proliferation.Western blot was used to detect Cyclin D1,MMP-2 and MMP-9 protein expression; Transwell assay was used to detect cell migration and invasion.And real-time PCR(RT-qPCR)was used to detect miR-494 expression levels.Results:Compared with the control group,Anemarrhena asphodeloides saponin AⅢ in the 8 μmol/L medium dose group and 16 μmol/L high dose group significantly down-regulated the cell A value,migration number and invasion number as well as Cyclin D1,MMP-2 and MMP-9 protein expression in the group(P<0.05).Compared with the control group,Anemarrhena asphodeloides saponin AⅢ at 8 μmol/L medium dose group and 16 μmol/L high dose group significantly up-regulated miR-494 expression levels in cells(P<0.05).Compared with the miR-NC group,cell A value,migration number and invasion number as well as Cyclin D1,MMP-2 and MMP-9 protein expression were significantly lower in the miR-494 group(P<0.05).Compared with the anti-miR-NC +16 μmol/L group,the miR-494 expression level was significantly decreased in the anti-miR-494 +16 μmol/L group,the cell A value,the number of migrations and invasions,and Cyclin D1,MMP-2 and MMP-9 protein expression were significantly increased(P<0.05).Conclusion:miR-494 up-regulation by Anemarrhena asphodeloides saponin A Ⅲ inhibits the proliferation,migration and invasion of primary cutaneous melanoma.

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备注/Memo

备注/Memo:
基金项目:湖北省卫生健康委员会科研项目(WJ2021M088)
更新日期/Last Update: 2022-10-09