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转录组谱的下一代测序数据与印度尼西亚鼻咽癌患者增殖异常有关。

Transcriptome Profile of Next-Generation Sequencing Data Relate to Proliferation Aberration of Nasopharyngeal Carcinoma Patients in Indonesia.

机构信息

Study Program of Biotechnology, Graduate School, Universitas Gadjah Mada, Yogyakarta, Indonesia.

Bioinformatics and Data Science Research Center, Bina Nusantara University, Jakarta, Indonesia.

出版信息

Asian Pac J Cancer Prev. 2020 Sep 1;21(9):2585-2591. doi: 10.31557/APJCP.2020.21.9.2585.

Abstract

OBJECTIVE

Nasopharyngeal carcinoma (NPC) is the most common cancer arising from epithelial cells of the nasopharynx in  Indonesia. This study aims to determine the differential level of gene expression in NPC patients when compared with normal individuals. Transcriptome profiling analysis was performed using RNA-Seq technology to determine the differential gene expression relate to proliferation aberration that occurs in NPC patients compared with normal individuals. So we get the transcriptomic profile of Indonesia NPC patients.

METHODS

In this study, we used 9 samples, 7 NPC samples and 2 normal samples as control.  Fresh tissue of tumor samples was collected from biopsy, and normal samples were collected brushing technique. The total RNA was isolated from fresh tissue samples and brushing samples using the Rneasy® RNA Extraction Mini Kit. The cDNA library was generated using TruSeq® RNA Library Preparation Kit V2, and its concentration was determined using qPCR. The library was sequenced using the Next-Generation Sequencing (NGS) Illumina Next Seq 550 platform. The raw sequence data quality was analyzed using FastQC and interpreted using HISAT2, HTSeq, edgeR, and PANTHER.

RESULTS

From the analysis, 25493 gene transcripts were expressed, with 1956 genes were significantly upregulated, 90 genes were significantly downregulated in NPC samples, and 23897 genes didn't change the expression level significantly (p <0.05), 10 of which genes were associated with cell proliferation. These genes are involved in the regulation of cancer cell proliferation through several signaling pathways, which are the apoptosis signaling pathway, IGF signaling pathway, Notch signaling pathway, and P13K signaling pathway.

CONCLUSION

There were significant differences in gene expression levels between NPC patients and normal individuals. Each gene that has changed the expression level plays a role in regulating various pathways that lead to cell proliferation aberration in NPC cases.

摘要

目的

鼻咽癌(NPC)是印度尼西亚鼻咽部上皮细胞最常见的癌症。本研究旨在确定 NPC 患者与正常个体相比基因表达的差异水平。使用 RNA-Seq 技术进行转录组谱分析,以确定与 NPC 患者中发生的增殖异常相关的差异基因表达。因此,我们获得了印度尼西亚 NPC 患者的转录组谱。

方法

在这项研究中,我们使用了 9 个样本,7 个 NPC 样本和 2 个正常样本作为对照。从活检中采集肿瘤样本的新鲜组织,并用刷拭技术采集正常样本。使用 Rneasy® RNA 提取试剂盒从新鲜组织样本和刷拭样本中分离总 RNA。使用 TruSeq® RNA 文库制备试剂盒 V2 生成 cDNA 文库,并使用 qPCR 确定其浓度。使用 Next-Generation Sequencing (NGS) Illumina NextSeq 550 平台对文库进行测序。使用 FastQC 分析原始序列数据质量,并使用 HISAT2、HTSeq、edgeR 和 PANTHER 进行解释。

结果

从分析中,表达了 25493 个基因转录本,其中 1956 个基因明显上调,90 个基因在 NPC 样本中明显下调,23897 个基因表达水平没有明显变化(p<0.05),其中 10 个基因与细胞增殖有关。这些基因通过几种信号通路参与调节癌细胞增殖,即凋亡信号通路、IGF 信号通路、Notch 信号通路和 P13K 信号通路。

结论

NPC 患者与正常个体之间的基因表达水平存在显著差异。每个改变表达水平的基因在调节导致 NPC 病例中细胞增殖异常的各种途径中都发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/121f/7779438/9d1fae5bca98/APJCP-21-2585-g001.jpg

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