Zhai Congying, Liu Baoliang, Kan Fanggong, Zhai Shuhui, Zhang Ronghua
Department of Pulmonary and Critical Care Medicine, Zibo First Hospital, Zibo, Shandong 255200, P.R. China.
Department of Oncology, Zibo First Hospital, Zibo, Shandong 255200, P.R. China.
Oncol Lett. 2023 Sep 28;26(5):492. doi: 10.3892/ol.2023.14079. eCollection 2023 Nov.
The present study aimed to investigate microRNA (miRNA)-27a-3p expression in the pulmonary macrophages and peripheral blood of patients with early non-small cell lung carcinoma (NSCLC) and its regulatory effect on the infiltration of pulmonary macrophages into cancer tissues and invasion of NSCLC cells. Blood specimens were withdrawn from 36 patients with NSCLC and 29 healthy subjects. NSCLC tissues and cancer-adjacent tissues were both obtained from patients with NSCLC; furthermore, certain tissue samples were used to extract macrophages. The levels of miRNA-27a-3p and C-X-C motif ligand chemokine 2 (CXCL2) mRNA were detected by reverse transcription-quantitative PCR and the levels of CXCL2 protein were measured by ELISA and western blot analysis. A dual-luciferase reporter assay was performed to determine the interactions between miRNA and mRNA. An MTT assay was employed to examine the viability of transfected cells and macrophages and a Transwell assay was performed to assess chemotaxis. The differential expression of miRNA-27a-3p in NSCLC tissues, pulmonary macrophages and peripheral blood indicated that miRNA-27a-3p exerted different roles in these specimens. CXCL2 was upregulated in NSCLC tissues at both transcriptional and translational levels. In addition, the untranslated region of CXCL2 was confirmed to be directly targeted by miRNA-27a-3p prior to its transcriptional activation. Furthermore, miRNA-27a-3p regulated CXCL2 expression, thereby affecting the proliferation of human pulmonary macrophages. The present study highlights that miRNA-27a-3p expression in the pulmonary macrophages and peripheral blood of patients with NSCLC is downregulated, while its target gene CXCL2 is upregulated. miRNA-27a-3p may regulate the viability and chemotaxis of macrophages in tumor tissues of patients with NSCLC through CXCL2 and is expected to become a genetic marker of this disease.
本研究旨在探讨微小RNA（miRNA）-27a-3p在早期非小细胞肺癌（NSCLC）患者肺巨噬细胞和外周血中的表达及其对肺巨噬细胞浸润至癌组织和NSCLC细胞侵袭的调节作用。从36例NSCLC患者和29名健康受试者中采集血样。NSCLC组织和癌旁组织均取自NSCLC患者；此外，取部分组织样本提取巨噬细胞。采用逆转录定量PCR检测miRNA-27a-3p和C-X-C基序配体趋化因子2（CXCL2）mRNA水平，采用酶联免疫吸附测定（ELISA）和蛋白质印迹分析检测CXCL2蛋白水平。进行双荧光素酶报告基因检测以确定miRNA与mRNA之间的相互作用。采用MTT法检测转染细胞和巨噬细胞的活力，采用Transwell法评估趋化性。miRNA-27a-3p在NSCLC组织、肺巨噬细胞和外周血中的差异表达表明其在这些样本中发挥不同作用。CXCL2在NSCLC组织的转录和翻译水平均上调。此外，在转录激活前，证实CXCL2的非翻译区是miRNA-27a-3p的直接靶标。此外，miRNA-27a-3p调节CXCL2表达，从而影响人肺巨噬细胞的增殖。本研究强调，NSCLC患者肺巨噬细胞和外周血中miRNA-27a-3p表达下调，而其靶基因CXCL2上调。miRNA-27a-3p可能通过CXCL2调节NSCLC患者肿瘤组织中巨噬细胞的活力和趋化性，有望成为该疾病的遗传标志物。
