THG-1/TSC22D4 promotes inflammatory IL-1/NF-?B pathway.

Malignant neoplasms arise within a region of chronic inflammation caused by tissue injuries. Inflammation is a key factor involved in all aspects of tumorigenesis including initiation, proliferation, invasion, angiogenesis, and metastasis. Interleukin-1 (IL-1) plays critical functions in tumor development with influencing the tumor microenvironment and promoting cancer progression. RNA sequencing analysis performed with HaCaT-HRasG12V cells, HaCaT-HRasG12V-THG-1 (WT) cells and HaCaT-HRasG12V THG-1 (S264A mutant) cells revealed that THG-1 overexpression enhances the transcription of NF-?B targets including IL1A, IL1B, TNFA, and IL8. Overall design: A human keratinocyte HaCaT cell line was used in this study. Oncogenic HRasG12V mutant was first expressed in HaCaT cells (HaCaT-Ras). The HaCaT-Ras cells were further transduced with empty vector (-), THG-1 wild type (WT), and THG-1 (S264A). As a result, HaCaT-Ras (-), HaCaT-Ras-THG-1 (WT), and THG-1 (S264A) cell lines were generated. Total RNA was extracted from the three cell lines cultured in DMEM containing 10% fetal bovine serum.

恶性肿瘤（Malignant neoplasms）可发生于组织损伤诱发的慢性炎症区域。炎症是肿瘤发生发展全过程的关键调控因素，涵盖肿瘤起始、细胞增殖、侵袭、血管生成及远处转移等各个环节。白细胞介素-1（Interleukin-1, IL-1）可通过调控肿瘤微环境、促进癌症进展，在肿瘤发生发展中发挥关键作用。本研究针对三株细胞——HaCaT-HRasG12V细胞、HaCaT-HRasG12V-THG-1（野生型，WT）细胞以及HaCaT-HRasG12V THG-1（S264A突变体）细胞开展RNA测序（RNA sequencing）分析，结果显示THG-1过表达可增强NF-κB（核因子κB）靶基因的转录水平，相关靶基因包括IL1A（白细胞介素1α）、IL1B（白细胞介素1β）、TNFA（肿瘤坏死因子α）及IL8（白细胞介素8）。总体实验设计：本研究选用人角质形成细胞HaCaT细胞系作为研究模型。首先在HaCaT细胞中过表达致癌性HRasG12V突变体，构建得到HaCaT-Ras细胞。随后将空载体、THG-1野生型（WT）以及THG-1（S264A）分别转导至HaCaT-Ras细胞中，最终获得三株稳定细胞系：HaCaT-Ras（空载体对照组）、HaCaT-Ras-THG-1（WT组）及THG-1（S264A突变体组）。将上述三株细胞置于添加10%胎牛血清的DMEM培养基中培养后，提取总RNA用于后续测序分析。