子宫颈癌淋巴结转移全转录组表达谱的分析与验证  

Analysis and verification of the whole Transcriptome expression profile of lymph node metastasis in Cervical cancer

作　　者：许星月 郭依琳 王璐[1] 李瑞[1] 胡桂明[2] 赵虎[1] XU Xingyue;GUO Yilin;WANG Lu;LI Rui;HU Guiming;ZHAO Hu(Department of Obstetrics and Gynecology,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China;Department of Pathology,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China)

机构地区：[1]郑州大学第二附属医院妇产科,河南郑州450014 [2]郑州大学第二附属医院病理科,河南郑州450014

出　　处：《肿瘤基础与临床》2024年第3期256-261,共6页journal of basic and clinical oncology

基　　金：河南省高等学校重点科研项目(24B320029)。

摘　　要：目的通过高通量测序及生信分析,挖掘子宫颈癌淋巴结转移调控机制。方法收集3例淋巴结转移、3例无淋巴结转移的子宫颈癌组织,全转录组高通量测序筛选差异表达信使RNA(mRNA)、微小RNA(miRNA)和长链非编码RNA(lncRNA)(|log 2FC|≥1、P<0.05),构建竞争内源性RNA(ceRNA)网络,进行基因本体(GO)和京都基因与基因组百科全书(KEGG)通路分析。癌症基因组图谱(TCGA)数据集转移相关mRNA与ceRNA网络中mRNA取交集,检测交集mRNA表达情况。结果高通量测序筛选出差异表达的118个mRNA(47个上调,71个下调)、5个miRNA(1个上调,4个下调)和64个lncRNA(35个上调,29个下调),构建22个mRNA、5个miRNA、13个lncRNA的ceRNA网络。GO分析发现22个mRNA主要富集在趋化因子调节、tRNA甲基转移酶活性等;KEGG通路分析发现22个mRNA主要富集于Wnt、Rap1、MAPK、TNF等信号通路。TCGA数据集筛选出1404个转移相关的mRNA,与22个mRNA取交集后得到TRMT9B、FRAS1、BEND7、SLC35G1,实时定量聚合酶链反应结果显示,相比于无淋巴结转移的子宫颈癌患者,淋巴结转移的子宫颈癌患者BEND7(Z=3.628,P<0.001)、FRAS1(Z=2.570,P=0.010)和TRMT9B(Z=3.024,P=0.002)mRNA相对表达量更低,SLC35G1(Z=1.965,P=0.049)mRNA相对表达量更高。免疫组化结果显示,BEND7表达与国际妇产科联合会分期、淋巴脉管间隙浸润和淋巴结转移有关(χ^(2)=17.500,P<0.001;χ^(2)=4.351,P=0.037;χ^(2)=17.500,P<0.001)。结论本研究描绘子宫颈癌淋巴结转移表达谱,构建ceRNA网络,筛选并验证4个子宫颈癌淋巴结转移相关分子,为深入研究子宫颈癌淋巴结转移分子机制提供了思路。Objective To explore the regulatory mechanism of cervical cancer lymph node metastasis through high-throughput sequencing and bioinformatics analysis.Methods The 3 cases of cervical cancer tissues with lymph node metastasis and 3 cases without lymph node metastasis were collected,high-throughput whole transcriptome sequencing was used to screen for differentially expressed messenger RNA(mRNA),microRNA(miRNA),and long chain non coding RNA(lncRNA)(|log 2FC|≥1,P<0.05),a competitive endogenous RNA(ceRNA)network was constructed,and gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis was used.the intersection of metastasis related mRNA in the Cancer Genome Atlas(TCGA)dataset and mRNA in the ceRNA network was taken,and the expression of the intersecting mRNA was detected.Results The 118 differentially expressed mRNA(47 upregulated,71 downregulated),5 miRNAs(1 upregulated,4 downregulated)and 64 lncRNAs(35 upregulated,29 downregulated)were screened by high throughput sequencing,and a ceRNA network of 22 mRNA,5 miRNAs and 13 lncRNAs was constructed.GO analysis revealed that 22 mRNA were mainly enriched in chemokine regulation and tRNA methyltransferase activity;KEGG pathway analysis revealed that 22 mRNA were mainly enriched in signaling pathways such as Wnt,Rap1,MAPK,and TNF.The 1404 transfer related mRNA were screened from the TCGA dataset and intersected with 22 mRNA to obtain TRMT9B,FRAS1,BEND7,SLC35G1.Real time quantitative polymerase chain reaction results showed that compared to cervical cancer patients without lymph node metastasis,BEND7(Z=3.628,P<0.001),FRAS1(Z=2.570,P=0.010)and TRMT9B(Z=3.024,P=0.002)had lower mRNA expression levels,while SLC35G1(Z=1.965,P=0.049)had higher mRNA expression levels in the cervical cancer patients with lymph node metastasis.The immunohistochemical results showed that BEND7 expression was associated with International Federation of Obstetrics and Gynecology staging,lymphatic vessel interstitial infiltration and lymph node metastasis(χ^(2)=17.500,P<0.0

关 键 词：子宫颈癌 全转录组高通量测序 淋巴结转移 竞争内源性RNA 表达谱 

分 类 号：R737.33[医药卫生—肿瘤]