江苏大学学报（医学版）2025，Vol.35Issue(5)：392-400,428,10.DOI:10.13312/j.issn.1671-7783.y240105
小鼠胰腺癌放疗联合免疫治疗的转录组测序分析及关键基因的筛选和验证
Transcriptome sequencing analysis of radiotherapy combined with immunotherapy for pancreatic cancer and screening and validation of key genes in mice
摘要
Abstract
Objective:To investigate the changes in gene expression in pancreatic cancer during radiotherapy combined with immunotherapy and to screen and validate key genes involved in mice.Methods:Twenty C57BL/6J male mice aged 6-8 weeks were randomly divided into 4 groups:control group,radiotherapy group,immunotherapy group and radiotherapy combined with immunotherapy group,with 5 mice in each group.The tumor-bearing mouse model was established by subcutaneous injection of pancreatic cancer KPC cells,and the radiotherapy group was irradiated with 8 Gy radiation,the immunotherapy group received a single intraperitoneal injection of PD-1 monoclonal antibody(200 μg/mouse),the radiotherapy combined with immunotherapy group received a single dose of 8 Gy radiation followed by intraperitoneal injection of PD-1 monoclonal antibody(200 μg/mouse),and the control group did not receive any treatment.The tumor tissues were harvested 7 days later,and the transcriptome sequencing was performed.The differentially expressed genes(DEGs)with significant expression were analyzed by GO,KEGG and Reactome functional annotation to explore the related signaling pathways.Venn analysis was performed to identify the intersection of DEGs,and survival analysis was used to identify key genes.The expression levels of these key genes were analyzed,and their correlation with immune cell infiltration and immune checkpoint expression was examined.A protein-protein interaction(PPI)network was constructed to analyze the interactions of these key genes.Two subcutaneous tumor-bearing models of male mice with pancreatic cancer KPC and PANC02 cell lines C57BL/6J were constructed.The combined radiotherapy and immunotherapy treatment was administered on the following day and maintained for 3 days.Quantitative reverse transcription PCR(qRT-PCR)was used to assess the relative mRNA expression of key genes in tumor tissues.Results:The results of transcriptome sequencing analysis showed that compared with the radiotherapy group and the immunotherapy group,the radiotherapy combined with immunotherapy group had the richest DEGs and mainly participated in the signal pathways related to metabolism,the immune system and signal transduction.Venn analysis of upregulated genes in the radiotherapy,immunotherapy,and combined radiotherapy and immunotherapy groups identified 11 common genes.Survival analysis revealed that myxovirus resistance 1(MX1)was a key gene,which was highly expressed in pancreatic cancer and significantly negatively correlated with the prognosis of pancreatic cancer patients(P<0.05).The MX1 mRNA expression in pancreatic cancer was positively correlated with the infiltration levels of neutrophils and dendritic cells(r=0.413,0.333,both P<0.05),and was also positively correlated with the expression of immune checkpoints CD80,CD274,and CD86(all P<0.05).The gene cluster interacting with MX1 in pancreatic cancer included OASL1,RSAD2,IFIT1,IFIT2 and IFIT3.qRT-PCR results showed that in the tumor-bearing mouse models of pancreatic cancer KPC and PANC02 cells,compared with the control group,the relative expression level of MX1 mRNA in the radiotherapy combined with immunotherapy group increased significantly(P<0.05).Conclusion:The relative expression of MX1 mRNA significantly increased in radiotherapy combined with immunotherapy for pancreatic cancer,and it may be one of the key genes.关键词
MX1/转录组测序/胰腺癌/放疗联合免疫治疗Key words
MX1/transcriptome sequencing/pancreatic cancer/radiotherapy combined with immunotherapy分类
医药卫生引用本文复制引用
陈双伟,任永飞,王玥,蔡华忠..小鼠胰腺癌放疗联合免疫治疗的转录组测序分析及关键基因的筛选和验证[J].江苏大学学报（医学版）,2025,35(5):392-400,428,10.基金项目
镇江市重点研发计划(社会发展)项目(SH2022042) （社会发展）
