CD1a promotes the proliferation of the lung adenocarcinoma cell line A549 by regulating the expression of CCR4

YAN Xingyu; LIU Hongyan

doi:10.3969/j.issn.1002-7386.2025.07.004

Abstract：Objective To investigate the role of abnormal expression of cluster of differentiation 1a(CD1a)in cell proliferation of lung adenocarcinoma and the underlying mechanism.Methods A549 cells were transfected with lentivirus.A549 cells were transfected with LV-NC(negative lentivirus),LV-CD1a(stable overexpression of CD1a),si-NC,si-CD1a-1 or si-CD1a-2.Moreover,A549 cells transfected with LV-NC or LV-CD1a were co-transfected with si-NC,si-CCR4-1,si-CCR4-2 or si-CCR4-3.The mRNA and protein levels of CD1a and C-C chemokine receptor 4(CCR4)were detected by quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot,respectively.Differentially expressed genes in A549 cells transfected with LV-NC or LV-CD1a were screened by Whole-transcriptome analysis with total RNA sequencing(RNA-Seq).Cell viability at 0h,24h,48h,72h,96h and 120h was assessed by MTT assay.Results Analysis of The Cancer Genome Atlas(TCGA)and Clinical Proteomic Tumor Analysis Consortium(CPTAC)databases showed that CD1a was significantly up-regulated in lung adenocarcinoma specimen(both P＜0.05).Among the 15 pairs of lung cancer tissue samples,up-regulated mRNA level of CD1a was detected in 10 lung adenocarcinoma than that of paired adjacent tissues(all P＜0.05).MTT assay showed that transfection of si-CD1a-2 significantly decreased the proliferation activity at 24h than those transfected with si-NC(P＜0.01).No significant difference in the proliferation activity was detected between A549 cells transfected with si-CD1a-1 and si-NC(P＞0.05).At 48h,72h,96h,and 120h,the proliferation activities of A549 cells transfected with si-CD1a-1 or si-CD1a-2 was significantly lower than those transfected with si-NC(P＜0.05).Meanwhile,the proliferation activity of A549 cells transfected with LV-CD1a was significantly higher than those transfected with LV-NC(P＜0.05).RNA-Seq data showed that CCR4 was differentially expressed in A549 cells transfected with LV-CD1a and LV-NC(t=-3.935,P＜0.05).qRT-PCR validated the significantly higher mRNA level of CCR4 in A549 cells transfected with LV-CD1a than those transfected with LV-NC A(t=-5.063,P＜0.01).Knockdown of CCR4 in A549 cells overexpressing CD1a(A549 cells co-transfected with LV-CD1a and si-CCR4-1,si-CCR4-2 or si-CCR4-3),MTT assay showed significantly reduced proliferation ability than those co-transfected with LV-CD1a and si-NC(all P＜0.05).Conclusion CD1a is overexpressed in lung adenocarcinoma and promotes the proliferation of the lung adenocarcinoma cell line A549 by regulating the expression of CCR4.