The clinical significance and potential molecular mechanism of PTTG1 in esophageal squamous cell carcinoma

Esophageal squamous cell carcinoma (ESCC) is the major histological type of esophageal cancers worldwide. Transcription factor PTTG1 was seen highly expressed in a variety of tumors and was related to the degree of tumor differentiation, invasion, and metastasis. However, the clinical significance of PTTG1 had yet to be verified, and the mechanism of abnormal PTTG1 expression in ESCC was not clear. In this study, we collected eight pairs of ESCC samples and corresponding adjacent normal esophageal tissues for RNA-seq from eight ESCC patients underwent radical resection of EC. None of these ESCC patients received preoperative chemotherapy or radiotherapy.The tissue samples were stored at -80°C immediately after resection. These 8 patients included 6 males and 2 females with a median age of 60 years. RNA-seq was executed by Wuhan Seqhealth Technology Co.LTD. TRIzol reagent (Invitrogen, Carlsbad, CA, USA) was applied to extract and prepare total RNA from tumors and paired adjacent non-ESCC tissues. A total amount of 3 μg RNA per sample was used as initial material for the RNA sample preparations. Nanodrop™ 1000 spectrophotometer was applied to examined RNA purity. RNA concentration was determined using a Qubit® 3.0 Fluorometer. RNA integrity was determined using an RNA Nano 6000 Assay Kit and a Bioanalyzer 2100 system. Afterward, the sequencing libraries were established using the NEBNext Ultra Directional RNA Library Prep Kit for Illumina base on manufacturer’s protocols. The 200 Illumina HiSeq X Ten platform was used to sequence the libraries. RNA sequencing reads were aligned to the reference of homo_sapiens.GRCh38. In the current study, we obtained the data on log2 (x + 0.001) conversion level 3 and reported RNA-seq data in transcripts per kilobase million (TPM). mRNA profiles of ESCC samples and corresponding adjacent normal esophageal tissues.

食管鳞状细胞癌（Esophageal squamous cell carcinoma, ESCC）是全球范围内食管癌的主要组织学类型。转录因子PTTG1在多种肿瘤中呈高表达状态，且与肿瘤分化程度、侵袭及转移密切相关。然而，PTTG1的临床意义尚待验证，其在ESCC中表达异常的分子机制尚未明确。本研究收集了8例行食管癌根治性切除术的ESCC患者的癌组织及对应癌旁正常食管组织，用于RNA测序。所有入组的ESCC患者均未接受术前化疗或放疗。样本于切除后立即置于-80℃冰箱保存。该8例患者中，男性6例、女性2例，中位年龄为60岁。RNA测序由武汉Seqhealth科技有限公司（Wuhan Seqhealth Technology Co.LTD）完成。采用TRIzol试剂（Invitrogen，美国加利福尼亚州卡尔斯巴德市）提取并制备肿瘤组织及配对癌旁非ESCC组织的总RNA。每个样本取3 μg总RNA作为RNA样品制备的起始材料。使用Nanodrop™ 1000分光光度计检测RNA纯度，采用Qubit® 3.0荧光定量仪测定RNA浓度，使用RNA Nano 6000检测试剂盒与Bioanalyzer 2100系统评估RNA完整性。随后，基于制造商提供的操作流程，使用NEBNext Ultra Directional RNA Library Prep Kit for Illumina构建测序文库。采用200 Illumina HiSeq X Ten平台对文库进行测序。将RNA测序读段比对至人类参考基因组homo_sapiens.GRCh38。本研究采用log₂(x + 0.001)转换后的第3层级数据，并以每百万转录本（transcripts per kilobase million, TPM）格式报告RNA测序数据。本数据集包含ESCC样本及对应癌旁正常食管组织的mRNA表达谱。