TWIST1 Activates CDCA3 to Promote EMT of Lung Adenocarcinoma and Expression of PD-L1

Abstract

CDCA3 and TWIST1 are implicated in cell-cycle control and transcriptional regulation, yet their combined role in lung adenocarcinoma (LUAD) remains unclear. Here, we investigated the impact of the TWIST1/CDCA3 axis on LUAD cell behavior. Leveraging TCGA and KnockTF v2.0, we pinpointed TWIST1 as an upstream driver of CDCA3 in LUAD and delineated associated survival outcomes. ChIP and dual-luciferase assays verified the interaction between CDCA3 and TWIST1. We also quantified CDCA3 and TWIST1 mRNA by qRT-PCR and determined CDCA3, the immune-therapy biomarker PD-L1, and EMT-related protein levels by Western blot. Furthermore, CCK-8, colony formation assays, wound healing, and Transwell assays were conducted to evaluate the malignant behaviors of cells. Bioinformatic and functional analyses revealed that both CDCA3 and TWIST1 were highly expressed in LUAD, and their elevated levels predicted poor prognosis. Further investigation identified TWIST1 as an upstream activator of CDCA3. Knockdown of TWIST1 attenuated the malignant phenotype and reduced PD-L1 expression in LUAD cells, whereas subsequent overexpression of CDCA3 fully reversed these suppressive effects. This study aims to validate that the TWIST1/CDCA3 axis promotes invasion, migration, proliferation, and PD-L1 expression of LUAD cells.