Analysis and verification of the whole Transcriptome expression profile of lymph node metastasis in Cervical cancer
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 me-tastasis 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)(|log2 FC|≥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 me-tastasis 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 1 404 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 immunohisto-chemical results showed that BEND7 expression was associated with International Federation of Obstetrics and Gynecology staging,lymphatic vessel interstitial infiltration and lymph node metastasis(χ2=17.500,P<0.001;χ2=4.351,P=0.037;χ2=17.500,P<0.001).Conclusion This study describes the expression profile of cervical cancer lymph node metastasis,constructs a ceRNA network,screens and validates four cervical cancer lymph node metastasis related mole-cules,provides ideas for in-depth research on the molecular mechanism of cervical cancer lymph node metastasis.
万方数据
维普
