背景:肿瘤相关巨噬细胞(TAMs)显著影响进展,转移,和食管鳞状细胞癌(ESCC)的复发。ESCC中长的非编码RNA(lncRNAs)的异常表达已经建立,然而,在ESCC进展期间,lncRNAs在TAM重编程中的作用仍未被研究.
方法:通过将差异表达的lncRNAs与免疫相关的lncRNAs交叉并进行免疫细胞浸润分析来鉴定ESCCTAM相关的lncRNAs。使用TCGA数据库和临床样品检查LINC00330的表达谱和临床相关性。构建LINC00330过表达和干扰序列以评估LINC00330对ESCC进展的影响。单细胞测序数据,CIBERSORTx,和GEPIA用于分析ESCC肿瘤微环境中的免疫细胞浸润,并评估LINC00330和TAM浸润之间的相关性。进行ESCC-巨噬细胞共培养实验以研究LINC00330对TAM重编程的影响及其对ESCC进展的后续影响。通过转录组测序证实了LINC00330与C-C基序配体2(CCL2)的相互作用,亚细胞定位分析,RNA下拉,银染,RNA免疫沉淀,和其他实验。
结果:LINC00330在ESCC组织中显著下调,与患者预后不良密切相关。LINC00330过表达抑制ESCC进展,包括扩散,入侵,上皮-间质转化,和体内致瘤性。LINC00330促进TAM重新编程,和LINC00330介导的TAM重编程抑制ESCC进展。LINC00330结合CCL2蛋白并抑制CCL2和下游信号通路的表达。CCL2对于LINC00330介导的TAM重编程和ESCC进展至关重要。
结论:LINC00330通过以自分泌方式破坏CCL2/CCR2轴及其下游信号通路,并以旁分泌方式阻碍CCL2介导的TAM重编程,从而抑制ESCC进展。由LINC00330/CCL2轴介导的TAM重编程新机制可能为ESCC患者的靶向和免疫联合治疗提供潜在策略。
BACKGROUND: Tumor-associated macrophages (TAMs) significantly influence the progression, metastasis, and recurrence of esophageal squamous cell carcinoma (ESCC). The aberrant expression of long noncoding RNAs (lncRNAs) in ESCC has been established, yet the role of lncRNAs in TAM reprogramming during ESCC progression remains largely unexplored.
METHODS: ESCC TAM-related lncRNAs were identified by intersecting differentially expressed lncRNAs with immune-related lncRNAs and performing immune cell infiltration analysis. The expression profile and clinical relevance of LINC00330 were examined using the TCGA database and clinical samples. The LINC00330 overexpression and interference sequences were constructed to evaluate the effect of LINC00330 on ESCC progression. Single-cell sequencing data, CIBERSORTx, and GEPIA were utilized to analyze immune cell infiltration within the ESCC tumor microenvironment and to assess the correlation between LINC00330 and TAM infiltration. ESCC-macrophage coculture experiments were conducted to investigate the influence of LINC00330 on TAM reprogramming and its subsequent effect on ESCC progression. The interaction between LINC00330 and C-C motif ligand 2 (CCL2) was confirmed through transcriptomic sequencing, subcellular localization analysis, RNA pulldown, silver staining, RNA immunoprecipitation, and other experiments.
RESULTS: LINC00330 is significantly downregulated in ESCC tissues and strongly associated with poor patient outcomes. Overexpression of LINC00330 inhibits ESCC progression, including proliferation, invasion, epithelial-mesenchymal transition, and tumorigenicity in vivo. LINC00330 promotes TAM reprogramming, and LINC00330-mediated TAM reprogramming inhibits ESCC progression. LINC00330 binds to the CCL2 protein and inhibits the expression of CCL2 and downstream signaling pathways. CCL2 is critical for LINC00330-mediated TAM reprogramming and ESCC progression.
CONCLUSIONS: LINC00330 inhibited ESCC progression by disrupting the CCL2/CCR2 axis and its downstream signaling pathways in an autocrine fashion; and by impeding CCL2-mediated TAM reprogramming in a paracrine manner. The new mechanism of TAM reprogramming mediated by the LINC00330/CCL2 axis may provide potential strategies for targeted and immunocombination therapies for patients with ESCC.