[Mechanisms of Periplaneta americana extract in regulating JAK1/PI3K/AKT signaling pathway to intervene in patient-derived early-stage lung adenocarcinoma organoids with ground-glass nodule features].

This study aimed to elucidate the therapeutic effects and molecular mechanisms of Periplaneta americana extract(PAE) on ground-glass nodule(GGN)-type early-stage lung adenocarcinoma(LUAD) using patient-derived organoid(PDO) models. GGN-type patient-derived early-stage LUAD organoid(GGN-PDO) were successfully established. At the level of drug quality control and component analysis, the content of free amino acids and peptides in PAE were determined by high-performance liquid chromatography(HPLC), and the main components were further analyzed by liquid chromatography-mass spectrometry(LC-MS). Analysis of variance was used to evaluate the similarity between drugs of the same batch(Pearson correlation coefficient 0.87-0.99) and ensure quality control. At the histopathological level, hematoxylin-eosin(HE) staining and immunohistochemical(IHC) analysis of key molecular markers, including cytokeratin 7(CK7), thyroid transcription factor-1(TTF-1), and Ki-67, confirmed that the organoids recapitulated the tumor characteristics of parental tissues. Pharmacodynamic evaluation demonstrated that PAE significantly inhibited the proliferation and viability of GGN-PDOs in a dose-dependent manner, while immunofluorescence(IF) staining showed a marked reduction in the proliferation marker Ki-67. Flow cytometry further confirmed that PAE induced apoptosis in GGN-PDO, and Western blot analysis demonstrated that PAE downregulated the anti-apoptotic protein Bcl-2 and upregulated the pro-apoptotic protein Bax, thereby activating the apoptotic pathway. Notably, for the first time, the pro-apoptotic mechanism of PAE was shown to involve inhibition of the Janus kinase 1(JAK1)/phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT) signaling pathway, as evidenced by markedly decreased protein levels of JAK1, PI3K, p-PI3K, AKT, and p-AKT following PAE treatment. In summary, this study is the first to use a clinically relevant GGN-PDO models to demonstrate that PAE induces apoptosis in early-stage LUAD via JAK1/PI3K/AKT pathway inhibition, providing a solid theoretical and experimental basis for developing innovative pharmacotherapies and non-surgical interventions against GGNs.