| Hepatocellular carcinoma（HCC）is a primary liver cancer,accounting for 95%of primary liver cancers.Globally,the incidence and lethality of hepatocellular carcinoma are increasing year by year.The symptoms of hepatocellular carcinoma are not obvious in the early stage,and most patients are found at an advanced stage,and there is a risk of recurrence and metastasis even after cure.The search for new biomarkers and molecular mechanisms of hepatocellular carcinoma is urgently needed.More and more studies have proved that m6 A methylation modification plays an important role in the occurrence and development of hepatocellular carcinoma.Exploring the molecular mechanism of m6 A methylation affecting the progression of hepatocellular carcinoma is of great significance for finding new prognostic markers and therapeutic targets for hepatocellular carcinoma.YTH domain family proteins are the recognition proteins of m6 A methylation modification.By recognizing RNAs modified by m6 A methylation,they participate in various metabolic processes of RNA,and affect the occurrence and development of tumors.However,the roles and molecular mechanisms of YTH domain family proteins in tumors need to be further explored.In this study,the role of YTH domain family proteins in pan-cancer was first explored,and it was found that YTH domain family proteins had an important correlation with hepatocellular carcinoma.Then,the role and mechanism of YTH domain in hepatocellular carcinoma were further explored.The result is as follows:1.Through bioinformatics research,it was found that YTH domain family proteins have important correlation with hepatocellular carcinoma.YTH domain family proteins were highly expressed in cholangiocarcinoma（CHOL）,gastric adenocarcinoma（STAD）and hepatocellular carcinoma（LIHC）.Comprehensive analysis of Kaplan-Meier survival analysis and univariate COX regression analysis showed that YTHDF1 was significantly associated with overall survival in hepatocellular carcinoma（LIHC）and uveal melanoma（UVM）.YTHDF2 was associated with survival in low-grade glioma（LGG）,hepatocellular carcinoma（LIHC）,clear cell renal cell carcinoma（KIRC）,and sarcoma（SARC）,and YTHDF3 was associated with survival in invasive breast cancer（BRCA）,clear cell renal cell carcinoma（KIRC）,acute myeloid leukemia（LAML）,and thyroid cancer（THCA）,YTHDC1 was associated with survival in bladder urothelial carcinoma（BLCA）,clear cell renal cell carcinoma（KIRC）and low-grade glioma（LGG）.YTHDC2 was associated with survival in low-grade glioma（LGG）,head and neck squamous cell carcinoma（HNSC）,malignant melanoma（SKCM）,and rectal adenocarcinoma（READ）.Combined with differential expression and survival analysis,it was concluded that YTH domain family proteins may play an important role in the occurrence and development of hepatocellular carcinoma.Other biological characteristics analysis results showed that YTH domain family proteins have important correlations with P13K/AKT signaling pathway and TSC/mTOR signaling pathway.YTH domain family proteins had strong sensitivity to anticancer drugs:nelalabine,chelerythrine and PX-316.2.YTHDF1 in YTH domain family protein could be used as an independent prognostic factor of hepatocellular carcinomaFirst,by analyzing the differential expression of 24 m6 A methylation related genes in hepatocellular carcinoma,we found that except for ZC3H13 and METTL14（P > 0.05）,the other 22 m6 A methylation related genes were highly expressed in hepatocellular carcinoma.The results of univariate COX regression survival analysis and multivariate COX regression survival analysis revealed that YTHDF1,YTHDF2 and KIAA1429 could construct a risk model,and the risk model could serve as an independent prognostic factor for hepatocellular carcinoma.In the GEPIA database,3 risk model genes were analyzed for overall survival and disease-free survival,and the results showed that YTHDF1 could serve as a prognostic core gene for hepatocellular carcinoma.3.YTHDF1 was highly expressed in hepatocellular carcinoma and could promote the proliferation of hepatocellular carcinoma.The normal liver cell line LO2 and liver cancer cell lines including HepG2,SMMC7721 and HUH7 were selected as research subjects.QRT-PCR and Western blot results proved that YTHDF1 was highly expressed in HepG2 cell line and SMMC7721 cell line.CCK8 assay and clonogenic assay illustrated that YTHDF1 could promote the proliferation and colony formation abilities of HCC.4.By analyzing the RNA-seq sequencing results,YHDF1 was found to promote the proliferation of hepatocellular carcinoma possibly through activating the Wnt signaling pathway.Knockdown of YTHDF1 in SMMC7721 cell line,the results of RNA-seq sequencing showed that compared with the control group,there were a total of 602 differential genes,including 384 downregulated genes and 218 upregulated genes.Genes without annotated gene names were then removed,resulting in a total of 490 differential genes with 310 downregulated and 180 upregulated genes.It was found that the differential genes were mainly enriched in PI3K/Akt signaling pathway,calcium signaling pathway,Ras signaling pathway,MAPK signaling pathway and Wnt signaling pathway.The MCODE algorithm was used to screen the top three key subnetworks with the score,and the results showed that the first subnetwork with the highest score was composed of 8 genes,including RASGRP1,CACNA1 E,CACNA1G,SCN3 B,RASGRP2,CACNA1 H,STAC3,and CACNB2.The second key subnetwork consisted of 10 genes,including TIMP3,CCL2,IGFBP3,WNT1,TGFB2,NOG,WNT11,WNT10 B,LEF1,and HNF4 A.The third key subnetwork consists of 9 genes,including GNG7,GNAT2,GNG4,GNB3,ARRHGEF6,PTGER3,PTGER4,VIP,and CHRM4.We found that the second key subnetwork contained 4 key genes,including Wnt1,Wnt10 b,Wnt11 and LEF1,illustrating that YTHDF1 is likely to promote the proliferation of hepatocellular carcinoma by activating the Wnt signaling pathway.In the present study,YTH domain family proteins were found to have important relevance to the development of hepatocellular carcinoma by analyzing their differential expression and survival correlation in Pan cancer.Further studies proved that YTHDF1 could serve as an independent prognostic factor in hepatocellular carcinoma and played an important role in the Wnt signaling pathway,MAPK signaling pathway,Ras signaling pathway and G protein coupled receptor signaling pathway.YTHDF1 may promote the proliferation of hepatocellular carcinoma by activating Wnt signaling pathway.Taken together,YTH domain family protein YTHDF1,as a recognition protein for m6 A methylation,plays an important role in the development of hepatocellular carcinoma. |
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