Exploring the Key Role of TBC1D16 in Lung Cancer Metastasis Based on Lung Cancer Organoids
Background: Lung cancer is a leading cause of cancer-related deaths globally, with metastasis being a major factor in poor prognosis. Understanding the molecular mechanisms of lung cancer metastasis is crucial for developing effective therapies. This study aims to utilize lung cancer organoids (LCOs) to screen for differentially expressed genes (DEGs) in lung cancer metastasis through transcriptome sequencing and to explore their key roles and regulatory mechanisms.Methods: Three LCOs were isolated and cultured, and metastatic LCOs were induced by TGF-beta. The organoids were identified by histological and immunofluorescence staining. Transcriptome sequencing was performed to identify DEGs between LCOs and LCOs treated with TGF-beta. TBC1D16 levels were further verified by qRT-PCR. Survival and ROC curve analyses were performed to evaluate the prognostic value of TBC1D16, and its correlation with tumor staging was analyzed. Lentiviral vectors overexpressing or interfering with TBC1D16 were constructed to study their effects on organoid activity, budding, and levels of epithelial-mesenchymal transition (EMT) markers (E-cadherin/N-cadherin).Results: Metastatic LCOs showed increased tumor budding, reduced E-cadherin, and elevated N-cadherin expression. Transcriptome analysis identified 628 DEGs, with TBC1D16 being significantly upregulated. TBC1D16 expression was associated with lung cancer prognosis, disease progression, and tumor staging. Overexpression of TBC1D16 enhanced the activity of LCOs, promoted budding, and induced changes in EMT markers, leading to the metastasis of LCOs, similar to the effects of TGF-beta treatment. Conversely, interference with TBC1D16 reduced the activity of LCOs, inhibited budding, reversed TGF-beta-induced EMT process, and suppressed the metastasis of LCOs.Conclusions: This study revealed the mechanism by which TBC1D16 promoted metastasis through the regulation of EMT in LCOs and confirmed its clinical prognostic value, providing a theoretical basis for targeted therapy of lung cancer.