Background: scriptional regulatory mechanism, intricately intertwined with tumorigenesis and its therapeutic interventions. Aberrant splicing has emerged as a prospective wellspring for targeted therapy in the context of tumour afflictions. However, limited investigations have been conducted concerning the nexus between AS and non-small cell lung cancer (NSCLC). In this study, we have uncovered a 147-bp splicing abnormality within the downstream target gene BTC, which is subject to regulation by the HNRNPDL gene in NSCLC. Nevertheless, the clinical significance of this splicing regulatory mechanism, as well as the splicing event itself, remains shrouded in obscurity.
Methods: Differential genes downstream of HNRNPDL, introduced into NSCLC cell lines through lentivirus-mediated short hairpin RNA (shRNA) transfection, were meticulously screened via RNA sequencing (RNA-SEQ). This yielded the identification of crucial splicing events associated with NSCLC. Expression variations in distinct spliceosomes within NSCLC cell lines were assessed through Western Blot and reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Furthermore, shRNA plasmids targeting various spliceosomes of HNRNPDL and BTC were delivered into NSCLC cell lines, instigating a series of in vitro experiments geared towards elucidating their biological functionalities.
Results: Expression profiling of HNRNPDL was up-regulation. Concurrently, RNA-seq analysis primarily engendered two transcripts, namely exon 4 (BTC-L) and exon 4 skipping transcripts (BTC-S), owing to exon 4 aberrancy. Notably, both BTC-L and BTC-S exhibited up-regulation in NSCLC cell lines when juxtaposed with normal bronchial cells (BEAS-2B). In vitro experiments unequivocally demonstrated that both BTC-L and BTC-S fostered proliferation, migration, and apoptosis in NSCLC, manifesting a congruent carcinogenic effect.
Conclusions: The HNRNPDL gene has been elucidated as a promoter of NSCLC occurrence and progression through its orchestration of BTC splicing abnormalities. This revelation serves as a cornerstone for the identification of potential targets for the focused therapy of NSCLC. Less...