Abstract：Objective: To investigate the effect of long non-coding RNA nuclear enriched abundant transcript 1 (LncRNA NEAT1) on the biological function of lung cancer A549 cells by regulating the miR-582-5p/COL5A1 signaling pathway.Methods: Human lung cancer A549 cells were cultured in vitro and randomly grouped into a control group, si-NEAT1 group (transfected with LncRNA NEAT1 siRNA plasmid), miR-582-5p mimics group (transfected with miR-582-5p mimics), si-NC+miR-582-5p-NC group (co transfected with LncRNA NEAT1 siRNA negative control and miR-582-5p negative control), and si-NEAT1+miR-582-5p inhibitor group (co transfected with LncRNA NEAT1 siRNA plasmid and miR-582-5p inhibitor). After grouping and transfection, real-time fluorescence quantitative PCR experiment was applied to detect the expression of LncRNA NEAT1, miR-582-5p, and COL5A1 in cells. Nude mouse transplantation tumor model was constructed, and the mass and volume of the nude mouse transplantation tumor were measured. CCK-8 experiment and clone formation experiment were applied to detect cell proliferation. Immunoblotting experiments were applied to detect the expression of proliferation related proteins (cyclin D1, PCNA) in cells and nude mouse transplanted tumor tissues. Transwell experiment was applied to detect cell migration and invasion. Immunoblotting experiment was applied to detect the expression of epithelial mesenchymal transition (EMT) related proteins (Vimentin, MMP2, E-cadherin) in cells. Dual luciferase reporter gene assay was applied to detect the targeted regulatory effects of LncRNA NEAT1 on miR-582-5p and miR-582-5p on COL5A1 in cells.Results: Compared with the control group, the expression of COL5A1 mRNA, cell viability and clone formation rate, weight and volume of nude mouse transplanted tumors, expression of cyclin D1 and PCNA proteins, numbers of cell migration and invasion, and expression of Vimentin and MMP2 proteins in cells and nude mouse transplanted tumor tissues reduced in the si-NEAT1 group and miR-582-5p mimics group (P<0.05), the expression of miR-582-5p and E-cadherin protein in cells increased (P<0.05); there was no great difference in all indicators in the si-NC+miR-582-5p-NC group (P>0.05). Compared with the si-NEAT1 group, the expression of COL5A1 mRNA, cell viability and clone formation rate, weight and volume of nude mouse transplanted tumors, expression of cyclin D1 and PCNA proteins, numbers of cell migration and invasion, and expression of Vimentin and MMP2 proteins in cells and nude mouse transplanted tumor tissues increased in the si-NEAT1+miR-582-5p inhibitor group (P<0.05), the expression of miR-582-5p and E-cadherin protein in cells decreased (P<0.05). LncRNA NEAT1 was able to target downregulation of miR-582-5p in A549 cells, and miR-582-5p was able to target downregulation of COL5A1 in A549 cells (P<0.05).Conclusion: Knocking down LncRNA NEAT1 can reduce COL5A1 expression by upregulating miR-582-5p expression, thereby inhibiting in vitro and in vivo growths of lung cancer A549 cells and weakening their invasion and migration activities.
陈志辉, 赵蒙, 屈鑫. LncRNA NEAT1靶向调控miR-582-5p/COL5A1信号通路对肺癌A549细胞生物学功能的影响[J]. 河北医学, 2024, 30(11): 1784-1791.
CHEN Zhihui, ZHAO Meng, QU Xin. Effect of LncRNA NEAT1 on the Biological Function of Lung Cancer A549 Cell through Targeted Regulation of the miR-582-5p/COL5A1 Signaling Pathway. HeBei Med, 2024, 30(11): 1784-1791.
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2024, Vol. 30 
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