上调miR-338-3p通过靶向MACC1抑制恶性黑色素瘤A375细胞增殖、迁移和侵袭  

Upregulation of miR-338-3p inhibits the proliferation,migration and invasion of malignant melanoma A375 cells by targeting MACC1

作　　者：覃璇 龙新纯[1] 王文娟[1] QIN Xuan;LONG Xinchun;WANG Wenjuan(Department of Dermatology,Chenzhou First People's Hospital,Chenzhou 423000,Hunan,China)

机构地区：[1]郴州市第一人民医院皮肤科,湖南郴州423000

出　　处：《中南医学科学杂志》2025年第2期226-231,共6页Medical Science Journal of Central South China

基　　金：湖南省郴州市科技计划项目(Jsyf2017039)。

摘　　要：目的 探讨上调miR-338-3p对恶性黑色素瘤A375细胞增殖、迁移和侵袭的影响机制。方法 纳入GSE8401、GSE18509黑色素瘤数据集和30例黑色素瘤组织标本,检测miR-338-3p、MET转录调节因子1(MACC1)在不同肿瘤组织中的表达情况。根据不同载体转染至A375细胞,细胞实验分为miR-NC组(转染miR-338-3p过表达空载体)、Vector组(转染MACC1过表达空载体)、sh-NC组(转染敲低MACC1空载体)、sh-MACC1组(转染敲低MACC1)、p-MACC1组(转染过表达MACC1)、miR-338-3p mimic组(转染miR-338-3p模拟物)及miR-338-3p mimic+p-MACC1组(转染miR-338-3p mimic和p-MACC1)。采用qRT-PCR评估miR-338-3p、MACC1 mRNA表达水平;Western blotting检测MACC1和B-Raf原癌基因(BRAF)/丝裂原活化蛋白激酶(MEK)/细胞外信号调控的蛋白激酶(ERK)通路相关蛋白的表达;采用克隆形成、Transwell和划痕愈合实验检测A375细胞增殖和迁移能力。双荧光素酶报告基因测定miR-338-3p和MACC1之间的靶向关系。裸鼠成瘤模型验证miR-338-3p和MACC1的作用。结果 在肿瘤转移组织和黑色素瘤细胞中,MACC1上调,miR-338-3p下调。miR-338-3p负调控MACC1表达。上调miR-338-3p、敲低MACC1抑制A375细胞增殖、侵袭、迁移和小鼠肿瘤的生长(P<0.05)。MACC1通过BRAF/MEK/ERK通路调控A375细胞恶性行为。结论 上调miR-338-3p抑制黑色素瘤A375细胞的增殖、迁移和侵袭,可能与MACC1调控BRAF/MEK/ERK信号通路有关。Aim To investigate the molecular mechanisms of miR-338-3p upregulation on proliferation,migration and invasion of malignant melanoma A375 cells.Methods GSE8401,GSE18509 melanoma datasets and 30 melanoma tissue specimens were included,and the expression of miR-338-3p,MET-associated in colon cancer 1(MACC1) in different tumour tissues were detected.A375 cells were transfected with different vectors and then divided into miR-NC group(transfected with miR-338-3p overexpression empty vector),Vector group(transfected with MACC1 overexpression empty vector),sh-NC group(transfected with knockdown MACC1 empty vector),sh-MACC1 group(transfected with knockdown MACC1),p-MACC1 group(transfected with overexpression MACC1),miR-338-3p mimic group(transfected with miR-338-3p mimic),and miR-338-3p mimic+p-MACC1 group(transfected with miR-338-3p mimic and p-MACC1).The expression levels of miR-338-3p and MACC1 mRNA in tumor tissues and cells were evaluated by qRT-PCR.Western blotting was used to detect the protein expression levels of MACC1,B-Raf proto-oncogene(BRAF)/mitogen-activated protein kinase(MEK)/extracellular signal-regulated kinase(ERK).Clone formation test,Transwell test and scratch healing test were used to determine the cell proliferation and metastasis ability.The targeting relationship between miR-338-3p and MACC1 was analyzed by dual luciferase reporter gene assay.Nude mouse tumor model was used to evaluate the relation of miR-338-3p and MACC1.Results MACC1 was upregulated,whereas miR-338-3p was downregulated in tumor metastatic tissues and melanoma cells.miR-338-3p negatively regulates MACC1 expression.Upregulation of microRNA-338-3p and knockdown of MACC1 inhibited the proliferation,invasion and migration of A375 cells and tumor growth in mice.MACC1 promoted the malicious behaviour of A375 cells by inhibiting the MEK/ERK pathway.Conclusion Upregulation of miR-338-3p inhibits the proliferation,migration and invasion of melanoma cells by targeting MACC1 to regulate BRAF/MEK/ERK signaling pathway.

关 键 词：miR-338-3p MACC1 黑色素瘤 BRAF/MEK/ERK通路 A375细胞 

分 类 号：R739.5[医药卫生—肿瘤]