Department of Head and Neck Surgery, Hunan Cancer Hospital & Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha 410013, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2023 May 28;48(5):633-640. doi: 10.11817/j.issn.1672-7347.2023.220316.
Tongue squamous cell carcinoma (TSCC) is a common cancer in the oral and maxillofacial region, which seriously endangers people's life and health.Heterogeneous nuclear ribonucleoprotein A2/B1(hnRNP A2/B1) is an RNA-binding protein that regulates the expression of a variety of genes and participates in the occurrence and development of a variety of cancers. This study aims to investigate the role of hnRNP A2/B1 in TSCC progression.
The differential expression of hnRNP A2/B1 in oral squamous cell carcinoma (OSCC) and normal oral mucosa cells and tissues was analyzed based on the gene expression profiles of GSE146483 and GSE85195 in the Gene Expression Omnibus (GEO) database. The correlation between hnRNP A2/B1 expression and disease-free survival of TSCC patients was analyzed based on TSCC related chip of GSE4676. TSCC cancer and paracancerous tissue samples of 30 patients were collected in Hunan Cancer Hospital from July to December 2021. Real-time RT-PCR and Western blotting were used to verify the mRNA and protein expression of hnRNP A2/B1 in TSCC patients'samples, respectively. Human TSCC Tca-8113 cells were transfected with hnRNP A2/B1 empty vector (a sh-NC group), knockdown plasmid (a sh-hnRNP A2/B1 group), empty vector overexpression plasmid (an OE-NC group) and overexpression plasmid (an OE-hnRNP A2/B1 group), respectively. The knockdown or overexpression efficiency of hnRNP A2/B1 was detected by Western blotting. The proliferation activity of Tca-8113 cells was detected by cell counting kit-8 (CCK-8), and the apoptosis rate of Tca-8113 cells was detected by flow cytometry.
Based on the analysis of OSCC-related chips of GSE146483 and GSE85195 in the GEO database, it was found that hnRNP A2/B1 was differentially expressed in the OSCC and normal oral mucosa cells and tissues (all <0.01). Meanwhile, the analysis of TSCC related chip GSE4676 confirmed that the expression of hnRNP A2/B1 was negatively correlated with the disease-free survival of TSCC patients (=0.006). The results of real-time RT-PCR and Western blotting showed that the relative expression levels of hnRNP A2/B1 mRNA and protein in TSCC tissues were significantly up-regulated compared with those in adjacent tissues (all <0.01). The results of Western blotting showed that the expression level of hnRNP A2/B1 in Tca-8113 cells was significantly inhibited or promoted after knockdown or overexpression of hnRNP A2/B1 (all <0.01). The results of CCK-8 and flow cytometry showed that inhibition of hnRNP A2/B1 expression in Tca-8113 cells reduced cell proliferation activity (<0.05) and increased cell apoptic rate (<0.01). Overexpression of hnRNP A2/B1 in Tca-8113 cells significantly increased cell proliferation (<0.05) and decreased cell apoptosis (<0.01).
HnRNP A2/B1 is a key factor regulating the proliferation and apoptosis of TSCC cells. Inhibition of hnRNP A2/B1 expression can reduce the proliferation activity of TSCC cells and promote the apoptosis of TSCC cells.
舌鳞状细胞癌（TSCC）是口腔颌面部常见的癌症，严重危害人们的生命健康。异质核核糖核蛋白 A2/B1（hnRNP A2/B1）是一种 RNA 结合蛋白，可调节多种基因的表达，并参与多种癌症的发生和发展。本研究旨在探讨 hnRNP A2/B1 在 TSCC 进展中的作用。
基于基因表达谱公共数据库（GEO）中 GSE146483 和 GSE85195 分析 hnRNP A2/B1 在口腔鳞状细胞癌（OSCC）和正常口腔黏膜细胞和组织中的差异表达。基于 GSE4676 的 TSCC 相关芯片分析 hnRNP A2/B1 表达与 TSCC 患者无病生存的相关性。收集 2021 年 7 月至 12 月湖南省肿瘤医院 30 例 TSCC 癌及癌旁组织样本。分别采用实时 RT-PCR 和 Western blot 验证 TSCC 患者样本中 hnRNP A2/B1 的 mRNA 和蛋白表达。分别用 hnRNP A2/B1 空载（sh-NC 组）、敲低质粒（sh-hnRNP A2/B1 组）、空载过表达质粒（OE-NC 组）和过表达质粒（OE-hnRNP A2/B1 组）转染人 TSCC Tca-8113 细胞，Western blot 检测 hnRNP A2/B1 的敲低或过表达效率。采用细胞计数试剂盒-8（CCK-8）检测 Tca-8113 细胞的增殖活性，流式细胞术检测 Tca-8113 细胞的凋亡率。
基于 GEO 数据库中 OSCC 相关芯片 GSE146483 和 GSE85195 的分析，发现 hnRNP A2/B1 在 OSCC 和正常口腔黏膜细胞和组织中的表达存在差异（均<0.01）。同时，TSCC 相关芯片 GSE4676 的分析证实 hnRNP A2/B1 的表达与 TSCC 患者的无病生存呈负相关（=0.006）。实时 RT-PCR 和 Western blot 结果显示，与相邻组织相比，TSCC 组织中 hnRNP A2/B1 mRNA 和蛋白的相对表达水平均显著上调（均<0.01）。Western blot 结果显示，hnRNP A2/B1 在 Tca-8113 细胞中的表达在敲低或过表达 hnRNP A2/B1 后明显受到抑制或促进（均<0.01）。CCK-8 和流式细胞术结果表明，抑制 Tca-8113 细胞中 hnRNP A2/B1 的表达可降低细胞增殖活性（<0.05），增加细胞凋亡率（<0.01）。过表达 Tca-8113 细胞中的 hnRNP A2/B1 可显著增加细胞增殖活性（<0.05），降低细胞凋亡率（<0.01）。
hnRNP A2/B1 是调节 TSCC 细胞增殖和凋亡的关键因素。抑制 hnRNP A2/B1 的表达可降低 TSCC 细胞的增殖活性，促进 TSCC 细胞的凋亡。
