微小RNA-338-3p通过ERBB2调节卵巢癌细胞的增殖、凋亡、迁移及侵袭

doi:10.3969/j.issn.1006-7795.2025.03.018

摘要/Abstract

摘要： 目的探究微小RNA（microRNA，miR）-338-3p靶向人表皮生长因子受体2（human epidermal growth factor receptor 2，ERBB2）对卵巢癌（ovarian cancer，OC）细胞生物学行为的影响。方法收集45例OC患者的癌组织及癌旁组织并检测miR-338-3p、ERBB2表达水平。体外培养A2780、SKOV3、OVCAR3、IOSE-80细胞，分析比较miR-338-3p、ERBB2表达，并分析miR-338-3p与ERBB2的靶向关系。选择A2780进行后续实验，分为A2780组（正常A2780细胞）；阴性对照（negative control，NC）组（A2780细胞转染miR-NC）；miR-338-3p组（转染miR-338-3p模拟物）；pcDNA3.1+miR-338-3p组（转染miR-338-3p模拟物+pcDNA3.1）；miR-338-3p+ERBB2组（转染miR-338-3p模拟物+pcDNA3.1-ERBB2）。细胞计数试剂盒（cell counting kit-8，CCK-8）法检测细胞增殖；流式细胞仪测定凋亡率；Transwell小室测定细胞侵袭和迁移，免疫印迹法测定ERBB2蛋白及增殖、凋亡相关蛋白表达。A2780细胞注射至裸鼠右腋窝，记为模型组、NC组、miR-338-3p组、pcDNA3.1+miR-338-3p组、miR-338-3p+ERBB2组，每组6只。饲喂6周后，处死小鼠取肿瘤组织，测量体积和质量。免疫组化法分析肿瘤组织中ERBB2表达。结果 OC组织及细胞系中miR-338-3p表达低于癌旁组织和IOSE-80细胞，ERBB2 mRNA及蛋白表达高于癌旁组织和IOSE-80细胞（P<0.05）。miR-338-3p和ERBB2存在靶向关系，miR-338-3p负调控ERBB2表达。转染miR-338-3p模拟物后A2780细胞增殖、侵袭和细胞迁移数、ERBB2、B淋巴细胞瘤2（B-cellymphoma-2，Bcl-2）、磷酸化细胞外调节蛋白激酶1/2（phosphorylation extracellular signal-regulated kinase 1/2，p-ERK1/2）、基质金属蛋白酶9（matrixmetalloproteinase-9，MMP-9）蛋白表达降低，细胞凋亡率和Bcl-2相关X蛋白（Bcl-2 associated X，bax）表达增加（P<0.05）。转染ERBB2过表达质粒可增加A2780细胞增殖、侵袭和细胞迁移数以及ERBB2、Bcl-2、p-ERK1/2、MMP-9表达，降低bax表达和细胞凋亡率（P<0.05）。与模型组、NC组比较，miR-338-3p组肿瘤质量、体积及肿瘤组织中ERBB2表达降低（P<0.05），与miR-338-3p组、pcDNA3.1+miR-338-3p组比较，miR-338-3p+ERBB2组肿瘤质量、体积及肿瘤组织中ERBB2表达增加（P<0.05）。结论 miR-338-3p可能靶向调控ERBB2，从而抑制OC细胞生长和凋亡等行为。

关键词: 微小RNA-338-3p, 人表皮生长因子受体2, 卵巢癌, 细胞生物学行为, 增殖, 迁移

Abstract: Objective To investigate the effect of microRNA (miR)-338-3p on the biological behavior of ovarian cancer (OC) cells by targeting human epidermal growth factor receptor 2 (ERBB2). MethodsThe expression levels of miR-338-3p and ERBB2 were collected from the cancerous and adjacent tissues of 45 OC patients. A2780, SKOV3, OVCAR3, and IOSE-80 cells were cultured in vitro, and the expression of miR-338-3p and ERBB2 was analyzed and compared, the targeting relationship between miR-338-3p and ERBB2 was also analyzed. A2780 cells were randomly separated into A2780 group (normal A2780 cells), negative control (NC) group (A2780 cells transfected with miR-NC); miR-338-3p group (transfected with miR-338-3p mimic), PcDNA3.1+miR-338-3p group (transfected with miR-338-3p mimic+pcDNA3.1), and PcDNA3.1-ERBB2 group (transfected with miR-338-3p mimic+pcDNA3.1-ERBB2). The proliferation, apoptosis, invasion, and migration of cells in each group were analyzed, and the expression of ERBB2 protein and proliferation and apoptosis related proteins were measured. A2780 cells were injected into the right axilla of nude mice, and were recorded as model group, NC group, miR-338-3p group, pcDNA3.1+miR-338-3p group, miR-338-3p+ERBB2 group, with 6 cells in each group. After 6 weeks of feeding, the mice were killed and the tumor tissue was taken, and the volume and quality were measured. The expression of ERBB2 in tumor tissues was analyzed by immunohistochemistry. Results The expression of miR-338-3p in OC tissues and cell lines was lower than that in adjacent tissues and IOSE-80 cells, while the expression of ERBB2 mRNA and protein was higher than that in adjacent tissues and IOSE-80 cells (P<0.05). There was a targeting relationship between miR-338-3p and ERBB2, and miR-338-3p negatively regulated ERBB2 expression. After transfection with miR-338-3p mimic, the numbers of cell proliferation, invasion and migration, the expression of ERBB2, B-cellymphoma-2 (Bcl-2), phosphorylation extracellular signal-regulated kinase 1/2 (p-ERK1/2), matrixmetalloproteinase-9 (MMP-9) proteins decreased, while the apoptosis rate and the expression of Bcl-2 associated X (bax) protein increased (P<0.05). Transfection of ERBB2 overexpression plasmid was able to increase the numbers of cell proliferation, invasion and migration, the expression of ERBB2, Bcl-2, p-ERK1/2, MMP-9 proteins, and increase the apoptosis rate and the expression of bax protein (P<0.05). Compared with model group and NC group, tumor mass, volume and ERBB2 expression in tumor tissues of miR-338-3p group were decreased (P<0.05). Compared with miR-338-3p group and pcDNA3.1+miR-338-3p group, ERBB2 expression in tumor tissues of miR-338-3p group was decreased (P<0.05). The tumor mass, volume and the expression of ERBB2 in tumor tissues of miR-338-3p+ERBB2 group were increased (P<0.05). Conclusion miR-338-3p may target and regulate ERBB2, thereby inhibiting the growth and apoptosis of OC cells.

Key words: micro RNA-338-3p, human epidermal growth factor receptor 2, ovarian cancer, cellular biological behavior, proliferation, transfer

中图分类号:

R737.31

靖芳, 靖超. 微小RNA-338-3p通过ERBB2调节卵巢癌细胞的增殖、凋亡、迁移及侵袭[J]. 首都医科大学学报, 2025, 46(3): 527-537.

Jing Fang, Jing Chao. Micro RNA-338-3p regulates proliferation, apoptosis, migration, and invasion of ovarian cancer cells through ERBB2[J]. Journal of Capital Medical University, 2025, 46(3): 527-537.

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