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The expression of microRNA-324-3p as a tumor suppressor in nasopharyngeal carcinoma and its clinical significance

机译:microRNA-324-3p在鼻咽癌中的表达及其临床意义

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Objective: This study aimed to determine the expression, clinical significance, and possible biologic function of microRNA-324-3p in nasopharyngeal carcinoma (NPC) tissues. Methods: In total, 54 NPC and 35 control tissues were collected. The correlation between miR-324-3p expression and the clinicopathologic characteristics was analyzed. A dual-luciferase reporter gene assay was employed to examine the predicted target gene of miR-324-3p. The miR-324-3p expression level in 5–8F cells was determined with quantitative reverse transcription-polymerase chain reaction following the transfection of miR-324-3p mimics and inhibitors. Cell proliferation and the percentage of apoptosis were measured with MTT and flow cytometry. Cell invasion ability was assessed by Transwell invasion assay. Results: Our results showed that miR-324-3p was downregulated in the NPC tissues. The expression level of miR-324-3p in poorly differentiated NPC was significantly reduced in comparison with that in well/moderately differentiated NPC. The expression level in clinical stages?III/IV was lower than that in clinical stages I/II. Moreover, the expression level of miR-324-3p was significantly lower in NPC patients with lymph node metastasis than that in NPC patients without lymph node metastasis. NPC patients with higher levels of miR-324-3p expression also demonstrated a longer survival time. Predictions from bioinformatics indicated the Hedgehog pathway transcription gene GLI3 as the target gene of miR-324-3p, and the dual-luciferase reporter assay showed that miR-324-3p is directly combined with the 3′-untranslated region of GLI3. The overexpression of miR-324-3p suppressed cell proliferation and invasion, and it enhanced apoptosis in 5–8F cells. Conclusion: miR-324-3p can act as a tumor suppressor in NPC cells by the negative regulation of GLI3 gene.
机译:目的:本研究旨在确定microRNA-324-3p在鼻咽癌(NPC)组织中的表达,临床意义以及可能的生物学功能。方法:共收集54个NPC和35个对照组织。分析了miR-324-3p表达与临床病理特征之间的相关性。采用双荧光素酶报告基因检测miR-324-3p的预测靶基因。转染miR-324-3p模拟物和抑制剂后,通过定量逆转录聚合酶链反应确定了5-8F细胞中的miR-324-3p表达水平。用MTT和流式细胞术测量细胞增殖和凋亡百分比。通过Transwell侵袭测定法评估细胞侵袭能力。结果:我们的结果表明,miR-324-3p在NPC组织中被下调。与高分化/中分化的NPC相比,miR-324-3p在低分化的NPC中的表达水平显着降低。临床III / IV期的表达水平低于临床I / II期。此外,在有淋巴结转移的NPC患者中,miR-324-3p的表达水平明显低于没有淋巴结转移的NPC患者。具有较高水平的miR-324-3p表达的NPC患者还表现出更长的生存时间。生物信息学的预测表明,刺猬通路转录基因GLI3是miR-324-3p的靶基因,双荧光素酶报告基因检测结果表明miR-324-3p直接与GLI3的3'-非翻译区结合。 miR-324-3p的过表达抑制细胞增殖和侵袭,并增强5-8F细胞的凋亡。结论:miR-324-3p可通过负调控GLI3基因在NPC细胞中发挥抑癌作用。

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