基于转录组测序探究C-MET表达在非小细胞肺癌中的免疫调控机制
收稿日期: 2023-06-23
网络出版日期: 2024-01-24
基金资助
国家自然科学青年基金项目(8200113358);广东省自然科学基金(2022A1515012400);广州市科技计划项目(202206010134)
RNA sequencing⁃based research on the immune regulation mechanism of C⁃MET in lung cancer
Received date: 2023-06-23
Online published: 2024-01-24
目的 通过转录组测序技术分析C-MET表达在非小细胞肺癌中的免疫调控机制。 方法 使用siRNA分子干扰技术将C-MET高表达肺腺癌细胞株（H1993）、肺鳞癌细胞株（EBC-1）的C-MET表达沉默，利用转录组测序技术检测C-MET沉默前后细胞差异表达的基因（DEGs），通过生物信息学分析挖掘出C-MET可能参与调控的免疫微环境信号通路及相关基因。最后使用人免疫细胞与H1993、EBC-1共培养技术验证C-MET对免疫因子（INF-γ、INF-β、CXCL-10）的影响。 结果 通过转录组测序技术共检测到505个DEGs，其中H1993的C-MET调控前后表达差异组的表达差异基因共有38个，上调的差异表达基因有24个，下调的差异表达基因有14个。EBC-1的C-MET调控前后表达差异组的表达差异基因共有467个，上调的差异表达基因有347个，下调的差异表达基因121个。差异基因的KEGG分析表明，C-MET表达可能通过白细胞介素（IL-17）信号通路、白细胞分化、细胞因子受体活性、细胞周期、细胞因子-细胞因子受体相互作用参与免疫细胞调节因子的调控。使用肺癌细胞与人免疫细胞共同培养技术验证C-MET对免疫因子分泌的影响，Rt-qPCR检测结果提示：与C-MET高表达组共培养的PBMC中干扰素（INF-γ）的mRNA转录水平是低表达组的77倍、CXCL-10的mRNA转录水平是低表达组的1.6倍，INF-β的mRNA转录水平是低表达组的2倍。 结论 C-MET表达可能通过IL-17信号通路、白细胞分化、细胞因子受体活性通路参与肿瘤周围免疫微环境调控。
徐越 , 张言斌 , 苏珊 . 基于转录组测序探究C-MET表达在非小细胞肺癌中的免疫调控机制[J]. 实用医学杂志, 2024 , 40(1) : 7 -12 . DOI: 10.3969/j.issn.1006-5725.2024.01.002
Objective To analyze the immune regulation mechanism of C-MET expression in non-small cell lung cancer by transcriptome sequencing technology. Methods The C-MET expression of lung adenocarcinoma cell line （H1993） and lung squamous cell carcinoma cell line （EBC-1） with high C-MET expression was silenced using siRNA molecular interference technology. The differentially expressed genes （DEGs） before and after C-MET silencing were detected using transcriptome sequencing technology. The signal pathways and related genes of the immune microenvironment in which C-MET may participate in regulation were excavated through bioinformatics analysis. Finally， the co-culture technique of human immune cells with H1993 and EBC-1 was used to verify the effect of C-MET on immune factors such as INF-γ， INF-β and CXCL-10. Results We detected 505 DEGs in total using transcriptome sequencing.There were 38 differentially expressed genes in the C-MET regulation group before and after H1993， 24 upregulated differentially expressed genes， and 14 downregulated differentially expressed genes， respectively. There are a total of 467 differentially expressed genes in the C-MET regulation group of EBC-1， 347 upregulated differentially expressed genes， and 121 downregulated differentially expressed genes， respectively. KEGG analysis of differential genes suggested that C-MET expression might participate in the regulation of immune cell regulatory factors through the IL-17 signaling pathway， white blood cell differentiation， cytokine receptor activity， cell cycle， cytokine receptor activity， and cytokine-cytokine receptor interaction. The effect of C-MET on immune factor secretion was verified using the co-culture technique of lung cancer cells and human immune cells， and the results of Rt-qPCR assay suggested，the mRNA transcriptional level of INF-γ in PBMC co-cultured with the C-MET high expression group was 77 times that of the low expression group， and the mRNA transcriptional level of CXCL-10 was 1.6 times that of the low expression group. The mRNA transcriptional level of INF--β was twice as high as that of the low expression group. Conclusion C-MET expression may participate in the regulation of tumor surrounding immune microenvironment through IL-17 signaling pathway， leukocyte differentiation， and cytokine receptor activity pathway.
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