| 覃璇,龙新纯,王文娟.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.[J].中南医学科学杂志.,2025,(2):226-231. |
| 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 |
| 投稿时间：2023-02-17 修订日期：2024-10-09 |
| DOI：10.15972/j.cnki.43-1509/r.2025.02.008 |
| 中文关键词: miR-338-3p MACC1 黑色素瘤 BRAF/MEK/ERK通路 A375细胞 [ |
| 英文关键词:miR-338-3p MACC1 melanoma BRAF/MEK/ERK pathway A375 cell |
| 基金项目:湖南省郴州市科技计划项目(Jsyf2017039) |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| AimTo investigate the molecular mechanisms of miR-338-3p upregulation on proliferation, migration and invasion of malignant melanoma A375 cells. MethodsGSE8401, 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. ResultsMACC1 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. ConclusionUpregulation of miR-338-3p inhibits the proliferation, migration and invasion of melanoma cells by targeting MACC1 to regulate BRAF/MEK/ERK signaling pathway. |
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