miR-423-3p通过下调CARNS1促进乳腺癌细胞恶性生物学行为  

miR-423-3p Promotes Malignant Biological Behavior in Breast Cancer Cells by Downregulating CARNS1

作　　者：李金洋 陈东旭 傅腾超 吴琍[1] 

机构地区：[1]青岛大学附属医院乳腺病诊疗中心,山东 青岛

出　　处：《临床医学进展》2024年第3期415-423,共9页Advances in Clinical Medicine

摘　　要：目的:探讨miR-423-3p对乳腺癌细胞恶性生物学行为的影响及作用机制。方法:体外培养人乳腺癌细胞MDA-MB-231、MDA-MB-468、MCF-7、BT-549及正常乳腺上皮细胞MCF-10A,采用实时荧光定量PCR (RT-qPCR)法和Western Blot方法,分别检测miR-423-3p及CARNS1 mRNA和蛋白相对表达量。选取MCF-7细胞,将细胞分为转染mimics NC组(A组)、转染miR-423-3p mimics组(B组)、转染inhibitors NC组(C组)、转染inhibitors组(D组)。在239T细胞中分别转染并分为CARNS1-3'-UTR-wt+miR-423-3p mimics (E组)、CARNS1-3'-UTR-wt-mimics NC (F组)、CARNS1-3'-UTR-mut+miR-423-3p mimics (G组)及CARNS1-3'-UTR-mut+mimics NC (H组)。采用RT-qPCR和Western Blot法,检测A~D组细胞miR-423-3p及CARNS1 mRNA和蛋白表达情况,采用CCK8法、细胞划痕实验、Transwell法和流式细胞术检测细胞增殖、迁移、侵袭和凋亡情况。在线数据库预测miR-423-3p与CARNS1基因3'非翻译区(3'-UTR)的互补结合位点,双荧光素酶报告基因实验验证预测结果。结果:RT-qPCR结果显示,MCF-7、MDA-MB-231、MDA-MB-468及BT-549细胞中miR-423-3p的相对表达量均明显高于正常乳腺上皮细胞MCF-10A (P 0.05)。RT-qPCR检测结果显示,B组细胞的miR-423-3p相对表达量明显高于A组(P < 0.05),而CARNS1 mRNA相对表达量明显低于A组(P < 0.05),D组细胞的miR-423-3p相对表达量明显低于C组(P < 0.05),而CARNS1 mRNA相对表达量明显高于C组(P < 0.05)。结论:miR-423-3p可通过靶向调控CARNS1促进乳腺癌细胞的恶性生物学行为。Objective: To investigate the effect and mechanism of miR-423-3p on the malignant biological be-havior of breast cancer cells. Methods: Human breast cancer cells MDA-MB-231, MDA-MB-468, MCF-7, BT-549 and normal breast epithelial cells MCF-10A were cultured in vitro, and the relative mRNA and protein expressions of miR-423-3p and CARNS1 were detected by real-time quantitative PCR (RT-qPCR) and Western Blot, respectively. MCF-7 cells were divided into mimics NC transfection group (group A), miR-423-3p mimics group (group B), transfection inhibitors NC group (group C), and transfection inhibitors group (group D). 239T cells were transfected and divided into CARNS1-3'-UTR-wt miR-423-3p mimics (group E), CARNS1-3'-UTR-wt-mimics NC (group F), CARNS1-3'-UTR-mut miR-423-3p mimics (group G) and CARNS1-3'-UTR-mut mimics NC (group H). The mRNA and protein expressions of miR-423-3p and CARNS1 in group A~D cells were detected by RT-qPCR and Western Blot, and the proliferation, migration, invasion and apoptosis of cells were detected by CCK8 method, cell scratch assay, Transwell method and flow cytometry. The comple-mentary binding sites of miR-423-3p and the 3 untranslated region (3'-UTR) of CARNS1 gene were predicted by an online database, and the prediction results were verified by double luciferase re-porter gene assays. Results: RT-qPCR results showed that the relative expression levels of miR-423-3p in MCF-7, MDA-MB-231, MDA-MB-468 and BT-549 cells were significantly higher than those in normal mammary epithelial cells MCF-10A (P 0.05). The results of RT-qPCR showed that the relative expression of miR-423-3p in group B was signifi-cantly higher than that in group A (P < 0.05), while the relative expression of mRNA in CARNS1 was significantly lower than that in group A (P < 0.05), the relative expression of miR-423-3p in group D was significantly lower than that in group C (P < 0.05), and the relative expression of mRNA in CARNS1 was significantly higher than that in group C (P < 0.05). Conclusion: miR-423-3p can pro-mote

关 键 词：乳腺癌 微小RNA miR-423-3p CARNS1 

分 类 号：R73[医药卫生—肿瘤]