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

doi:10.3969/j.issn.1006-5725.2024.01.002

摘要/Abstract

摘要：

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。

关键词: 非小细胞肺癌, C-MET, 免疫微环境, 表达谱测序

Abstract:

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.

Key words: non-small cell lung cancer, C-MET, immune microenvironment, expression profile sequencing

中图分类号:

R734.2

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.

Yue XU,Yanbin ZHANG,Shan. SU. RNA sequencing⁃based research on the immune regulation mechanism of C⁃MET in lung cancer[J]. The Journal of Practical Medicine, 2024, 40(1): 7-12.

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参考文献 21

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基因名称	log₂FC	P值
DLX4	1.405	< 0.05
DDIT3	1.087	< 0.001
IRF5	1.145	< 0.01
EGR1	-2.280	< 0.001
RORC	1.196	< 0.001
ELF3	1.332	< 0.001
AFF1	1.311	< 0.001
ESX1	-1.622	< 0.01
ETV1	-1.915	< 0.001
ETV4	-1.585	< 0.001

基因名称	log₂FC	P值
PROX2	6.993	< 0.001
ZSCAN4	6.716	< 0.001
ZFP2	5.731	< 0.001
ZEB2	5.716	< 0.01
TBX1	5.261	< 0.01
ZNF880	5.100	< 0.01
TUB	4.898	< 0.001
SIM1	4.836	< 0.05
GATA1	4.814	< 0.05
ZNF410	4.708	< 0.01

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