SP1-mediated WTAP exacerbates the progression of colon adenocarcinoma by m6A methylation of KLK8
Zhijin Wu, Na Shen, Phil Hyun Song, Ben Wang, Chunming Li
Abstract
Background The study focused on the functions of kallikrein related peptidase 8 (KLK8), WT1 associated protein (WTAP) and specific protein 1 (SP1) in colon adenocarcinoma (COAD) development. Methods EdU and Transwell experiments were applied for cell proliferation, migration and invasion. Trypan blue experiment and flow cytometry analysis were adopted for evaluation of CD8 + T cell viability and apoptosis. qRT-PCR, western blot and IHC analysis were adopted to examine gene expression. The animal tests were utilized to investigate KLK8 role in tumor progression in vivo . meRIP analysis was conducted to estimate the relation between KLK8 and WTAP. CHIP and dual-luciferase reporter assays were performed to verify the relation between SP1 and WTAP. Results KLK8 expression was raised in COAD. Silencing of KLK8 inhibited COAD cell proliferation, migration, invasion, accelerated CD8 + T cell viability and repressed CD8 + T cell apoptosis. Deficiency of KLK8 also blocked tumor growth in vivo . WTAP modulated KLK8 expression through m6A methylation. WTAP silencing restrained COAD cell proliferation, migration, invasion and facilitated CD8 + T cell cytotoxicity, while KLK8 overexpression restored the effects. In addition, SP1 activated the transcription of WTAP. Deficiency of SP1 restrained COAD cell growth, metastasis and enhanced CD8 + T cell toxicity, with KLK8 enhancement abolished the effects. Conclusion SP1-mediated upregulation of WTAP exacerbated the malignant COAD progression by modulating KLK8.