甲基化修饰抑癌基因MSX1抑制黑色素瘤细胞迁移和侵袭的研究

doi:10.11904/j.issn.1002-3070.2020.06.006

实用肿瘤学杂志 ›› 2020, Vol. 34 ›› Issue (6): 511-517.doi: 10.11904/j.issn.1002-3070.2020.06.006

甲基化修饰抑癌基因MSX1抑制黑色素瘤细胞迁移和侵袭的研究

王灿¹, 罗茜¹, 罗焱¹, 刘素桃², 余音², 刁庆春², 黎智², 李晶²

1.重庆大学附属肿瘤医院;肿瘤转移与个体化诊治转化研究重庆市重点实验室(重庆 400030);
2.重庆市中医院皮肤科

收稿日期:2020-05-14 修回日期:2020-10-21 出版日期:2020-12-28 发布日期:2020-12-23
通讯作者: 李晶,E-mail:544746574@qq.com
作者简介:王灿,男,(1987-),博士,主治医师,从事肿瘤放化疗、肿瘤临床与基础的研究。
基金资助:
重庆市自然科学基金面上项目(编号:cstc2020jcyj-msxmX0355,cstc2020jcyj-msxmX0745)

Inhibitory migration and invasion of melanoma cells by methylation modified tumor suppressor gene MSX1

WANG Can¹, LUO Qian¹, LUO Yan¹, LIU Sutao², YU Yin², DIAO Qingchun², LI Zhi², LI Jing²

1. Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment,Chongqing University Cancer Hospital,Chongqing 400030,China;
2. Department of Dermatology,Chongqing Traditional Hospital

Received:2020-05-14 Revised:2020-10-21 Online:2020-12-28 Published:2020-12-23

摘要/Abstract

摘要： 目的探讨MSX1在黑色素瘤细胞系和组织中表达和甲基化及对黑色素瘤细胞A375迁移/侵袭的影响和机制。方法 qRT-PCR和免疫组织化学法分别检测MSX1在黑色素瘤细胞系和组织中的表达;甲基化PCR分析MSX1在黑色素瘤组织中甲基化;分别转染pcDNA3.1(+)-Flag-MSX1质粒(实验组)和pcDNA3.1(+)-Flag-Vector质粒(对照组)至A375细胞;Transwell迁移和侵袭实验检测MSX1对细胞迁移和侵袭的影响;RT-PCR验证MSX1对上皮间质转化及下游相关标志物的影响,免疫蛋白印迹实验验证MSX1对WNT/β-catenin信号通路的影响。结果与色素痣组织和细胞相比,MSX1 mRNA和蛋白在黑色素瘤细胞和组织中表达下调(P＜0.001);MSX1甲基化水平表达上调,其甲基化程度明显高于癌旁组织和正常色素痣组织。过表达MSX1后抑制细胞迁移和侵袭(P＜0.001),而上皮间质转化及下游相关标志物表达受到抑制。过表达MSX1能抑制WNT/β-catenin信号通路及其下游细胞因子的表达。结论 MSX1在黑色素瘤组织中表达下调,能抑制Wnt/β-catenin信号通路参与黑色素瘤的进展。

关键词: 黑色素瘤, MSX1, 甲基化, 抑癌基因, 迁移, 侵袭

Abstract: Objective The Objective of this study was to investigate the expression and methylation of MSX1 in melanoma cells and tissues,and its effect on migration/invasion of melanoma A375 cells. Methods qRT-PCR and immunohistochemical were used to examine the expression of MSX1 in A375 cells and tissues;methylation PCR was used to examine the methylation of MSX1 in melanoma tissues;pcDNA3.1(+)-Flag-MSX1(experimental group)and pcDNA3.1(+)-Flag-Vector(control group)were transfected into A375 cells,transwell migration and invasion assay were used to detect the effect of MSX1 on cell migration and invasion;RT-PCR was used to verify the effect of MSX1 on epithelial-mesenchymal transition and its downstream related markers;Westernblot was used to verify the effect of MSX1 on Wnt/β-catenin signaling pathway. Results MSX1 mRNA were down-regulated in human malignant melanoma cell lines and tissues(P＜0.001).The methylation of MSX1 was up-regulated,which was significantly higher than that in adjacent tissues and normal nevus tissues.Overexpression of MSX1 inhibited cell migration and invasion in A375 cells(P＜0.001),while epithelial to mesenchymal transition and the expression of downstream markers were also inhibited.MSX1 overexpressed could inhibit the expression of Wnt/β-catenin signaling pathway and its downstream markers in A375 cells. Conclusion MSX1 was down-regulated in melanoma tissues,indicating that MSX1 inhibits Wnt/β-catenin signaling pathway and participates in the progression of melanoma.

Key words: Melanoma, MSX1, Methylation, Tumor suppressor genes, Migration, Invasion

中图分类号:

R739.5

王灿, 罗茜, 罗焱, 刘素桃, 余音, 刁庆春, 黎智, 李晶. 甲基化修饰抑癌基因MSX1抑制黑色素瘤细胞迁移和侵袭的研究[J]. 实用肿瘤学杂志, 2020, 34(6): 511-517.

WANG Can, LUO Qian, LUO Yan, LIU Sutao, YU Yin, DIAO Qingchun, LI Zhi, LI Jing. Inhibitory migration and invasion of melanoma cells by methylation modified tumor suppressor gene MSX1[J]. Journal of Practical Oncology, 2020, 34(6): 511-517.

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链接本文: http://journal09.magtechjournal.com/Jwk3_syzlxzz/CN/10.11904/j.issn.1002-3070.2020.06.006

http://journal09.magtechjournal.com/Jwk3_syzlxzz/CN/Y2020/V34/I6/511

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甲基化修饰抑癌基因MSX1抑制黑色素瘤细胞迁移和侵袭的研究

Inhibitory migration and invasion of melanoma cells by methylation modified tumor suppressor gene MSX1

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