RPS27 选择性调节甲状腺癌细胞中炎症和免疫反应基因的表达和选择性剪接。

RPS27 selectively regulates the expression and alternative splicing of inflammatory and immune response genes in thyroid cancer cells.

机构信息

Ultrasound Department, People's Hospital of Xinjiang Uygur Autonomous Region, China.

Pathology Department, People's Hospital of Xinjiang Uygur Autonomous Region, China.

出版信息

Adv Clin Exp Med. 2022 Aug;31(8):889-901. doi: 10.17219/acem/147271.

DOI:10.17219/acem/147271

PMID:35546566

Abstract

BACKGROUND

The expression of ribosomal protein S27 (RPS27) is upregulated in multiple human malignancies. In thyroid cancer, the expression of RPS27 is associated with patient outcomes. However, the carcinogenic mechanisms of RPS27 and functions of RPS27 in the initiation and progression of thyroid cancer are still not clear.

OBJECTIVES

To investigate the carcinogenic mechanisms of RPS27 and functions of RPS27 in the initiation and progression of thyroid cancer.

MATERIAL AND METHODS

The RPS27 gene was overexpressed in BTH101 cells and the influence on the level of gene expression and alternative splicing (AS) was then analyzed by comparing the transcriptomes of the overexpressing cells with the controls. The procedures included cloning and plasmid construction of RPS27, cell culture and transfection, evaluation of RPS27 overexpression, library preparation and sequencing, RNA-Seq raw data clean and alignment, differentially expressed genes (DEGs) analysis, AS analysis, quantitative real-time polymerase chain reaction (qRT-PCR) validation of DEGs and AS events (ASEs), and functional enrichment analysis.

RESULTS

The results demonstrated that RPS27 could selectively regulate the expression of genes associated with autoimmune thyroid disease, inflammatory/immune response and AS of genes associated with TRIF-dependent toll-like receptor signaling pathway and apoptotic process. The genes in question are BMP6, SERPINA3, IL17B, IL1RN, HLA-B, PF4, HLA-DOB, MADCAM1, HLA-DQA1, TPO, HLA-B, HLA-DQA1, HLA-DOB, HLA-C, KRT8, CFLAR, HMGA1, CASP8, CCNH, UBE2D3, and MAPK9, among others.

CONCLUSIONS

The RPS27 selectively regulated the expression and alternative splicing of genes involved in inflammatory/immune response and TRIF-dependent toll-like receptor signaling pathway, which were tightly associated with the initiation and progression of thyroid cancer. These results extend our knowledge on the molecular functions of RPS27 in thyroid cancer cells and have a potential value in thyroid cancer treatment.

摘要

背景

核糖体蛋白 S27（RPS27）的表达在多种人类恶性肿瘤中上调。在甲状腺癌中，RPS27 的表达与患者预后相关。然而，RPS27 的致癌机制以及 RPS27 在甲状腺癌发生和进展中的功能仍不清楚。

目的

探讨 RPS27 在甲状腺癌发生和进展中的致癌机制及功能。

材料与方法

在 BTH101 细胞中过表达 RPS27 基因，通过比较过表达细胞和对照细胞的转录组，分析 RPS27 对基因表达和选择性剪接（AS）水平的影响。实验步骤包括 RPS27 基因的克隆和质粒构建、细胞培养和转染、RPS27 过表达的评估、文库制备和测序、RNA-Seq 原始数据的清洁和比对、差异表达基因（DEGs）分析、AS 分析、DEGs 和 AS 事件（ASEs）的定量实时聚合酶链反应（qRT-PCR）验证、以及功能富集分析。

结果

结果表明，RPS27 可以选择性调节与自身免疫性甲状腺疾病、炎症/免疫反应相关的基因的表达，以及与 TRIF 依赖性 Toll 样受体信号通路和凋亡过程相关的基因的 AS。涉及的基因有 BMP6、SERPINA3、IL17B、IL1RN、HLA-B、PF4、HLA-DOB、MADCAM1、HLA-DQA1、TPO、HLA-B、HLA-DQA1、HLA-DOB、HLA-C、KRT8、CFLAR、HMGA1、CASP8、CCNH、UBE2D3 和 MAPK9 等。