| Objective To explore the expression of genes related to hepatocellular carcinoma(HCC)stem cells and their prognostic correlation.Methods(1)Firstly,the transcriptome sequencing(RNA-seq)and clinical data of hepatocellular carcinoma were downloaded from the public database TCGA,and the mRNA expression-based stiffness index(mRNAsi)table of cancer stem cells was downloaded and sorted out to analyze the relationship between mRNAsi and pathological grade and prognosis of liver cancer.The mRNAsi of HCC was download and the prognostic value of mRNAsi was discussed.(2)Use the weighted gene coexpression network to screen the key modules related to LSCS from the differential genes of liver cancer,intercept the hub genes of the key modules.Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes were used for the functional and pathway enrichment analysis for hub genes.Through the online database STRING Construction of a hub gene protein-protein interaction(PPI)network by STRING and screening of key genes.(3)The key genes were analyzed for expression differences and expression correlations.(4)The online database The Human Protein Atlas was used to find and count the immunohistochemical staining results of key gene-encoded proteins in HCC tumor tissues and normal liver tissues to verify the key gene expression differences.(5)The k-m plotter database survival analysis and HCCDB data set validation results showed that 10 key genes were correlated with the prognosis of liver cancer patients.The overall survival time(Overall Survival,OS)in the high expression group was significantly lower than that in the low expression group(P <0.05).(6)A total of 44 HCC tissue samples were collected for immunohistochemical staining to further verify the expression of the key gene CCNB1 in HCC and adjacent tissues and its correlation with clinical pathological factors.Results(1)mRNAsi was significantly upregulated in cancer tissues(P <0.001),and increased with the increase of pathological grade of hepatocellular carcinoma(P=0.006).The mortality rate of the higher mRNAsi group was higher than that of the lower mRNAsi group(P=0.006).(2)The WGCNA network screened the blue module as the LCSC-related module,from which 24 hub genes were intercepted.Gene Ontology analysis found that hub genes were mainly involved in biological processes such as mitotic sister chromatid segregation?nuclear chromosome segregation nuclear division,and DNA conformation change and KEGG showed that hub genes were enriched in cell cycle,DNA mismatch repair,oocyte meiosis and other signaling pathways.PPI results showed that there was a strong interaction between the proteins encoded by the hub gene(P <0.001).(3)We screened 10?key genes(CCNB1?CDC20?CDCA8?NDC80?KIF20A?TTK?CDC45?KIF15?MCM2?NCAPG)related to mRNAsi of HCC based on WGCNA.The key genes were highly expressed in the tumor samples compared to the normal samples.In addition,there is a strong interaction between proteins of these key genes and a strong co-expression relationship at the transcriptional level(correlation coefficients were all?0.70).(4)The Human Protein Atlas identifies key genes that are cancer-related genes and are highly expressed in HCC tissues(lack of NCD80 data)(5)The k-m plotter database survival analysis and HCCDB data set validation results showed that 10 key genes were correlated with the prognosis of liver cancer patients.The overall survival time(Overall Survival,OS)in the high expression group was significantly lower than that in the low expression group(P<0.05).(6)Immunohistochemical results confirmed that CCNB1 was highly expressed in HCC and was closely correlated with tumor size,tumor number,presence or absence of cirrhosis,and Ki67 expression(P<0.05),and had no significant correlation with other pathological indicators(AFP,differentiation degree,TNM stage,presence or absence of portal vein tumor thrombi,etc.)(P>0.05).Conclusions(1)mRNAsi plays an important role in the occurrence and development of HCC.Based on WGCNA,10 key genes related to HCC-mRNAsi(CCNB1,CDC20,CDCA8,NDC80,KIF20 A,CDC45,KIF15,MCM2,TTK and NCAPG)are screened in LSCS.They may play an important role in the maintenance of stemness,and are expected to be therapeutic targets and prognostic markers for inhibiting the characteristics of HCC stem cells.(2)The high expression of CCNB1 in HCC tissues is related to the tumor size,number of tumors,presence or absence of cirrhosis,and Ki67 expression percentage of HCC,suggesting that CCNB1 may play an important role in the progression and metastasis of hepatocellular carcinoma,and CCNB1 may become an important molecule in HCC Markers and potential targets for the treatment of hepatocellular carcinoma. |
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