[1]韩昭,王久江,张伶,等.microRNA-136基于Wnt信号通路在黑素瘤EMT中的作用[J].中国皮肤性病学杂志,2020,(02):128-133.[doi:10.13735/j.cjdv.1001-7089.201906014]
　HAN Zhao,WANG Jiujiang,ZHANG Ling,et al.Role of MicroRNA-136 in EMT of Melanoma Based on Wnt Signaling Pathway[J].The Chinese Journal of Dermatovenereology,2020,(02):128-133.[doi:10.13735/j.cjdv.1001-7089.201906014]
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microRNA-136基于Wnt信号通路在黑素瘤EMT中的作用
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《中国皮肤性病学杂志》[ISSN:1001-7089/CN:61-1197/R]

卷:
期数:
2020年02期
页码:
128-133
栏目:
论著
出版日期:
2020-02-01

文章信息/Info

Title:
Role of MicroRNA-136 in EMT of Melanoma Based on Wnt Signaling Pathway
文章编号:
1001-7089(2020)02-0128-06
作者:
韩昭; 王久江; 张伶; 张建忠; 牛亮; 刘钊; 徐艳艳; 刘志军; 李小静
河北工程大学附属医院皮肤科,河北 邯郸 056000
Author(s):
HAN Zhao; WANG Jiujiang; ZHANG Ling; ZHANG Jianzhong; NIU Liang; LIU Zhao; XU Yanyan; LIU Zhijun; LI Xiaojing
(Department of Dermatology,Affiliated Hospital of Hebei University of Engineering,Handan 056000,China)
关键词:
microRNA-136;  黑素瘤;  Wnt信号通路;  EMT
Keywords:
MicroRNA-136;  Melanoma;  Wnt signaling pathway;  EMT
分类号:
R 739.5
DOI:
10.13735/j.cjdv.1001-7089.201906014
文献标志码:
A
摘要:
目的 探讨microRNA-136(miR-136)基于Wnt信号通路在黑素瘤上皮-间充质细胞转化(EMT)中的调控作用。方法 常规培养正常小鼠A9细胞(阴性对照组)和小鼠黑素瘤B16细胞,采用脂质体转染法将B16细胞分为空白组(只转染试剂,无转染序列)、NC组(转染miR-136 nc)、miR-136模拟组(转染miR-136模拟物)、miR-136抑制组(转染miR-136抑制剂)。通过qRT-PCR法检测各组细胞中miR-136的表达,qRT-PCR及Western blot实验分别检测各组细胞Wnt3a、β-catenin、E-cadherin及N-cadherin mRNA及蛋白表达的差距。结果 与阴性对照组相比,其他各组miR-136表达均显著降低,空白组与NC组比较差异无统计学意义(P>0.05)。相对于空白组与NC组,miR-136模拟组miR-136表达显著增加(P<0.05); miR-136抑制组miR-136的表达明显降低(P<0.05)。与对照组相比,其他各组E-cadherin mRNA及蛋白表达显著降低(P均<0.05)。β-catenin、Wnt3a、N-cadherin的mRNA及蛋白表达显著增加(P均<0.05)。空白组与NC组比较差异无统计学意义(P>0.05)。与空白组和NC组相比,miR-136模拟组E-cadherin的mRNA及蛋白表达显著增加(P<0.05); β-catenin、Wnt3a和N-cadherin的mRNA及蛋白表达均显著降低(P均<0.05)。miR-136抑制组E-cadherin的mRNA及蛋白表达显著降低(P均<0.05),而β-catenin、wnt3a和N-cadherin的mRNA及蛋白表达显著升高(P均<0.05),组间差距均具有统计学意义。结论 黑素瘤细胞中miR-136水平较正常细胞降低,过度表达的miR-136通过下调Wnt信号传导途径,抑制黑素瘤细胞的EMT。
Abstract:
Objective To explore the regulatory role of microrna-136(miR-136)in melanoma EMT based on the Wnt signaling pathway.Methods Normal mouse A9 cells(negative control group)and B16 mouse melanoma cells were cultured routinely.B16 cells were divided into the blank group(no transfection sequence),the NC group(transfected with miR-136 NC),the miR-136 simulation group(transfected with miR-136 mimics)and the miR-136 inhibition group(transfected with miR-136 inhibitors)by liposome transfection.The expression of miR-136 in each group was detected by qRT-PCR.The differences in mRNA and protein expression of Wnt3a,β-catenin,E-cadherin and N-cadherin in each group were detected by qRT-PCR and western-blot respectively.Results Compared with the negative control group,the expression of miR-136 in other groups was significantly reduced,and there was no significant difference between the blank group and the NC group(P>0.05).Compared with the blank group and the NC group,the expression of miR-136 in the miR-136 simulation group was significantly increased(P<0.05).The expression of miR-136 was significantly decreased in the miR-136 inhibition group(P<0.05).Compared with the control group,the mRNA and protein expressions of E-cadherin in other groups were significantly reduced(all P<0.05).The mRNA and protein expressions of β-catenin,Wnt3a,and N-cadherin were significantly increased(all P<0.05).There was no significant difference between the blank group and the NC group(P>0.05).Compared with the blank group and NC group,mRNA and protein expressions of E-cadherin in miR-136 simulation group were significantly increased(all P<0.05),whereas those of β-catenin,Wnt3a and N-cadherin were significantly decreased(all P<0.05).The mRNA and protein expressions of E-cadherin in the miR-136 inhibitor group were significantly decreased(all P<0.05),while those of β-catenin,Wnt3a and N-cadherin were significantly increased(all P<0.05).The differences between groups were statistically significant.Conclusion The level of miR-136 in melanoma cells is lower than that in normal cells.Over-expression of miR-136 inhibits EMT in melanoma cells by down-regulating Wnt signal transduction pathway.

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备注/Memo

备注/Memo:
[作者单位] 河北工程大学附属医院皮肤科,河北 邯郸 056000
[通信作者] 李小静,E-mail:zlmdsh@126.com[Corresponding author] LI Xiaojing,E-mail:zlmdsh@126.com
更新日期/Last Update: 2020-02-28