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miR-302c-3p 和 miR-520a-3p 通过靶向 CXCL8 抑制宫颈癌细胞的增殖。

miR‑302c‑3p and miR‑520a‑3p suppress the proliferation of cervical carcinoma cells by targeting CXCL8.

机构信息

Department of Obstetrics and Gynaecology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China.

Department of Medical Engineering, Suzhou Municipal Hospital, Suzhou, Jiangsu 215008, P.R. China.

出版信息

Mol Med Rep. 2021 May;23(5). doi: 10.3892/mmr.2021.11961. Epub 2021 Mar 24.

Abstract

The aim of the present study was to identify the differentially expressed microRNAs (miRs) in cervical carcinoma (CC) tissues and cells and to explore the function of miR‑302c‑3p and miR‑520a‑3p in the proliferation of CC cells. Potential dysregulated miRNAs in CC tissues and tumour‑adjacent tissues were detected. Reverse transcription‑quantitative PCR (RT‑qPCR) was performed to determine the expression of miR‑302c‑3p, miR‑520a‑3p and CXCL8 in CC tissues and cell lines. The target genes of the miRNAs were predicted using miRTarBase and verified by luciferase reporter assays. RT‑qPCR and western blotting were performed to measure the expression of C‑X‑C motif ligand (CXCL)8 after transfection. The effect on proliferation was verified by Cell Counting Kit assay and ethynyl‑2‑deoxyuridine staining. Flow cytometry was utilised to assess the effect on apoptosis. In the present study, miR‑302c‑3p and miR‑520a‑3p were markedly downregulated in CC cell lines compared to the normal cervical cell line H8. Functionally, overexpression of miR‑302c‑3p and/or miR‑520a‑3p inhibited proliferation and promoted the apoptosis of CC cell lines , while the knockdown of miR‑302c‑3p and/or miR‑520a‑3p had the opposite effect. Furthermore, miR‑302c‑3p and miR‑520a‑3p could both bind to CXCL8. Inhibition of CXCL8 in combination with miR‑302c‑3p and/or miR‑520a‑3p overexpression exerted proliferation‑suppressive and apoptosis‑stimulating effects on CC cells, whereas restoring CXCL8 attenuated the miR‑302c‑3p‑ and miR‑520a‑3p‑induced anti‑proliferative and pro‑apoptotic effects. miR‑302c‑3p and miR‑520a‑3p suppress the proliferation of CC cells by downregulating the expression of CXCL8, which may provide a novel target for the treatment of CC.

摘要

本研究旨在鉴定宫颈癌组织和细胞中差异表达的 microRNAs (miRs),并探讨 miR-302c-3p 和 miR-520a-3p 对宫颈癌细胞增殖的作用。检测宫颈癌组织和肿瘤旁组织中潜在失调的 miRNAs。采用逆转录定量 PCR (RT-qPCR)检测 miR-302c-3p、miR-520a-3p 和 CXCL8 在宫颈癌组织和细胞系中的表达。利用 miRTarBase 预测 miRNA 的靶基因,并通过荧光素酶报告基因实验进行验证。采用 RT-qPCR 和 Western blot 检测转染后 CXCL8 的表达。通过细胞计数试剂盒和溴脱氧尿苷掺入实验检测增殖的影响。采用流式细胞术检测凋亡的影响。在本研究中,与正常宫颈细胞系 H8 相比,miR-302c-3p 和 miR-520a-3p 在宫颈癌细胞系中明显下调。功能上,过表达 miR-302c-3p 和/或 miR-520a-3p 抑制增殖并促进宫颈癌细胞系的凋亡,而敲低 miR-302c-3p 和/或 miR-520a-3p 则产生相反的效果。此外,miR-302c-3p 和 miR-520a-3p 均可与 CXCL8 结合。抑制 CXCL8 联合 miR-302c-3p 和/或 miR-520a-3p 过表达对宫颈癌细胞发挥增殖抑制和促凋亡作用,而恢复 CXCL8 则减弱 miR-302c-3p 和 miR-520a-3p 诱导的抗增殖和促凋亡作用。miR-302c-3p 和 miR-520a-3p 通过下调 CXCL8 的表达抑制宫颈癌细胞的增殖,这可能为宫颈癌的治疗提供新的靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd82/7974325/7ec08b33052e/mmr-23-05-11961-g00.jpg

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