基于转录组测序探究有氧运动改善结直肠癌相关性肌少症的潜在干预机制

Exploring the potential intervention mechanism of aerobic exercise in improving colorectal cancer-associated sar⁃copenia based on RNA-sequence

目的基于转录组测序(RNA-sequence,RNA-seq)挖掘有氧运动对结直肠癌相关性肌少症改善作用的潜在干预机制。方法将20只5周龄BALB/C雄性裸鼠随机分为运动组与对照组。运动组跑步适应3 d后进行皮下瘤接种并进行为期5周的跑步训练,对照组不进行相关跑步训练,其他因素均同运动组,干预结束后采集裸鼠体重、瘤体数据,并进行腓肠肌组织RNA-seq分析。结果运动组的瘤重(P<0.01)、瘤体积(P<0.01)均显著低于对照组,而体质量显著高于对照组(P<0.05)。RNA-seq结果显示,运动组与对照组相比,差异基因共25个,其中24个为上调基因。上调基因蛋白质互作网络分析结果显示,Cxcl12、Ctla4、Flt3l等基因可能是关键靶标基因。基因本体论富集分析结果显示,上调条目共9个,其中“cytokine activity”条目的基因数量以及Log10(P)排名第一,相关基因包括Ccl3、Cxcl12、Cxcl2等。京都基因与基因组百科全书富集分析结果显示,上调条目共3个,“cytokine-cytokine receptor interaction”条目的基因数量以及Log10(P)排名第一,相关基因有Tnfsf18、Tnfrsf9、Tnfrsf11b等。RNA-seq上调差异基因与结直肠癌相关性肌少症靶标基因的交集基因共3个,包括Cxcl12、Ar、Mfge8。结论有氧运动可以抑制结直肠癌模型裸鼠的肿瘤生长,并且维持裸鼠体重增长,有效地防止荷瘤裸鼠肌少症以及恶病质情况的发生,其干预机制可能是多途径的,Cxcl12、Ar等均可能是有氧运动防治结直肠癌相关性肌少症的靶标基因。

Objective To explore the potential intervention mechanisms of aerobic exercise in improving colorectal cancer(CRC)-associated sarcopenia based on RNA-sequence(RNA-seq).Methods Twenty 5-week-old BALB/C male nude mice were randomly divided into an exercise group and a control group.After adapting to running for 3 days,the exercise group received subcutaneous tumor inoculation and underwent a 5-week running training.The control group did not receive any running training,while the rest were the same as the exercise group.After the end,the body weight and tumor data of nude mice were collected,and RNA-seq analysis of gastrocnemius muscle tissue was performed.Results The tumor weight(P<0.01)and tumor volume(P<0.01)of the exercise group were lower than those of the control group,while the body weight was higher(P<0.05).The RNA-seq showed that there was a total of 25 differentially expressed genes in the exercise group compared to the control group,of which 24 were upregulated genes.The analysis of upregulated gene protein-protein interaction(PPI)showed that genes such as Cxcl12,Ctla4,and Flt3l may be key target genes.The results of Gene Ontology(GO)enrichment analysis showed that there was a total of 9 upregulated entries,among which the number of genes and Log10(P)in the"cytokine activity"entry ranked first,and related genes included Ccl3,Cxcl12,Cxcl2,etc.The Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis results showed that there was a total of 3 upregulated entries,among which the number of genes and Log10(P)in the"cytokine-cytokine receptor interaction"entry ranked first,and related genes included Tnfsf18,Tnfrsf9,Tnfrsf11b,etc.There were three intersection genes between RNA-seq upregulated differentially expressed genes and CRC-associated sarcopenia target genes,including Cxcl12,Ar,and Mfge8.Conclusions Aerobic exercise can inhibit tumor growth in CRC model nude mice,maintain weight gain,and effectively prevent the occurrence of sarcopenia and cachexia in tumor bearing nude mice.The intervention mechanism may be multifaceted,and Cxcl12,Ar,etc.may be target genes for aerobic exercise to prevent CRC-associated sarcopenia.