Effects of microRNA-874-3p on biological behavior of lung adenocarcinoma cells through targeted regulation of plakophilin 3 and its mechanism

doi:10.16098/j.issn.0529-1356.2025.02.009

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Acta Anatomica Sinica ›› 2025, Vol. 56 ›› Issue (2) : 188-201. DOI: 10.16098/j.issn.0529-1356.2025.02.009

Effects of microRNA-874-3p on biological behavior of lung adenocarcinoma cells through targeted regulation of plakophilin 3 and its mechanism

CHEN Fan¹ TENG Zhao-hu¹ FANG Tao² REN Jun-xu³ ZHANG Jing^3* LI Xue¹ WANG Yi-xuan¹ LIN Xu³ WU Jing-fang¹

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Abstract

Objective The study aims to investigate the impact of microRNA-874-3p (miR-874-3p) regulation of plakophilin 3(PKP3)on the malignant biological behavior of lung adenocarcinoma cells and its underlying mechanism. Methods Immunohistochemistry and immunocytochemistry were used to detect the expression of PKP3 in lung adenocarcinoma tissue microarray and lung adenocarcinoma cell line A549 cells respectively, and the relationship between PKP3 and clinicopathological features of lung adenocarcinoma patients was analyzed. Select lung adenocarcinoma cell line A549, the experiment was divided into A549 cell group (blank control group), miR-NC group (transfected with miR-NC) and miR-mimics group (transfected with miR-874-3p mimics), sh-NC group (control group transfected with PKP3 silencing plasmid), sh-PKP3 group (transfected with PKP3 silencing plasmid), miR+pcDNA-PKP3 group (transfected with miR-874-3P mimics+pcDNA-PKP3, rescue group) and miR+pcDNA-NC group (transfected with miR-874-3p mimics+pcDNA-NC). The proliferation, invasion, migration and apoptosis of cells in each group were detected. ENCORI database was used to predict the upstream gene of PKP3, and dual luciferase assay was used to detect the targeting relationship between miR-874-3p and PKP3. MAPK/mTOR pathway-related proteins were detected by Western blotting. Results The expression of PKP3 in lung adenocarcinoma tissue was significantly higher than that in adjacent tissues. The high expression of PKP3 was related to clinical stage, tumor size, and lymph node metastasis (P<0.05).Compared with the human normal lung epithelial cells (BEAS-2B), the expression of PKP3 in A549 cells increased significantly, and the expression of miR-874-3p decreased (P<0.05). Overexpression of miR-874-3p decreased the PKP3 expression level (P<0.05). Compared with the control group, both overexpression of miR-874-3p and silenced PKP3 inhibited the cloning and invasion ability of A549 cells, caused cell cycle arrest, and decreased the expression levels of cyclin dependent kinase 4(CDK4), cyclin D1, cyclin E1 proteins in A549 cells (P<0.05). The expressions of Bax protein and Caspase-3 protein were up-regulated (P<0.05), and apoptosis increased. Overexpression of PKP3 could reverse the biological behavior of overexpression of miR-874-3p. Overexpression of miR-874-3p and silencing of PKP3 significantly decreased the expressions of P38 MAPK and mTOR phosphorylated proteins. Conclusion MiR-874-3p can negatively regulate PKP3 expression and inhibit the malignant biological behavior of A549 cells through MAPK/mTOR pathway.

Key words

MicroRNA-874-3p / / Plakophilin 3 / Lung adenocarcinoma / Proliferation / Invasion / Western blotting / Human

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CHEN Fan TENG Zhao-hu FANG Tao REN Jun-xu ZHANG Jing LI Xue WANG Yi-xuan LIN Xu WU Jing-fang. Effects of microRNA-874-3p on biological behavior of lung adenocarcinoma cells through targeted regulation of plakophilin 3 and its mechanism[J]. Acta Anatomica Sinica. 2025, 56(2): 188-201 https://doi.org/10.16098/j.issn.0529-1356.2025.02.009

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