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通过 CRISPR 敲除 miR-214 对 HPV 阳性和 HPV 阴性宫颈癌细胞的差异转录组分析。

Differential transcriptome analysis in HPV-positive and HPV-negative cervical cancer cells through CRISPR knockout of miR-214.

机构信息

Cancer Biology Lab, School of Biotechnology, KIIT University, Bhubaneswar, India.

出版信息

J Biosci. 2020;45.

Abstract

In this study we have investigated the effects of a tumour suppressor microRNA, miR-214, on gene expression in HPV-positive (CaSki) and HPV-negative cervical cancer cells (C33A) by RNA sequencing using next generation sequencing. The HPV-positive and HPV-negative cervical cancer cells were either miR-214- knocked-out or miR-214-overexpressed. Gene expression analysis showed that a total of 904 genes were upregulated and 365 genes were downregulated between HPV-positive and HPV-negative cervical cancer cells with a fold change of +/- 2. Furthermore, 11 differentially expressed and relevant genes which showed a fold change of +/-5 were selected to confirm by real-time PCR. This study represents the first report of miR-214 on global gene expression in the context of HPV.

摘要

在这项研究中,我们通过下一代测序的 RNA 测序,研究了肿瘤抑制 microRNA miR-214 对 HPV 阳性(CaSki)和 HPV 阴性宫颈癌细胞(C33A)基因表达的影响。我们通过 miRNA 敲除或过表达的方法,使 HPV 阳性和 HPV 阴性的宫颈癌细胞分别 miR-214 敲除或过表达。基因表达分析显示,HPV 阳性和 HPV 阴性宫颈癌细胞之间共有 904 个基因上调,365 个基因下调,差异倍数为 +/-2。此外,我们还选择了 11 个差异表达且差异倍数为 +/-5 的相关基因,通过实时 PCR 进行验证。本研究首次报道了 miR-214 在 HPV 背景下对全基因表达的影响。

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