Study On Identification,Validation And Associated Functions Of Cuproptosis-related LncRNA Signatures As A Novel Prognostic Model For Head And Neck Squamous Cell Cancer

Objective(s)Head and neck squamous cell cancer(HNSCC)is a common malignant cancer.We aimed to explore cuproptosis-related lncRNAs(CRLs)and establish CRL-related prognostic risk models for HNSCC.Method(s)Part I: The transcriptome profiles and clinical data were obtained from the TCGA database,and 19-cuproptosis-related genes(CRGs)were acquired from previous studies.Then,the prognostic model based on seven CRLs was established.We analyzed the correlation between the CRL-related prognostic risk models and the prognosis,drug sensitivity,and tumor immune functions of patients with HNSCC.Immunohistochemistry was applied to test the expression levels of the tumor microenvironment molecules(PD-1,PD-L1,PD-L2,CD3,CD4 and CD8)in nasopharyngeal squamous cell carcinoma.Finally,we used quantitative reverse transcription polymerase chain reaction(q RT-PCR)to validate the RNA expression levels of the seven CRLs.Part Ⅱ: We targeted WDFY3-AS2 for the gene loss-and-gain function cell experiment.q RT-PCR was used to verify the knockdown or overexpression efficiency of WDFY3-AS2 in CNE1 and Fa Du cells respectively;then,CCK-8assay was applied to test cell proliferation activity,Transwell chamber experiment was performed to detect cells migration and invasion ability.Western blot was used to detect the protein expression levels of E-cadherin,N-cadherin and Vimentin.Result(s)Part I:1.We established a 7-CRL signature.Kaplan-Meier(K–M)curve analysis demonstrated a significantly better prognosis in the low-risk group.Multivariate Cox regression analysis revealed that the risk score could serve as an independent prognostic factor.Nomogram,ROC curve,and principal component analysis indicated that the signature presented significant predictive capability.Moreover,most of the high-risk group showed lower levels of IC50 for certain chemotherapy drugs,such as cisplatin,cytarabine,docetaxel,doxorubicin,etoposide,gemcitabine,methotrexate,paclitaxel,and dasatinib.2.Weak response to immune checkpoint inhibitor was observed in patients with Nasopharyngeal squamous cell carcinoma,which may be partly attributable to its cold tumor microenvironment.3.The RNA expression levels of AP001372.2,MIR9-3HG,AL160314.2,POLH-AS1,and AL109936.2 were upregulated,while AC090587.1 and WDFY3-AS2 were downregulated in HNSCC cell lines compared with normal cell lines.Part Ⅱ:After the expression level of WDFY3-AS2 was inhibited,the proliferation efficacy of CNE1 and Fa Du cells were significantly promoted,and the ability of cell immigration and invasion was significantly enhanced;the protein expression level of E-cadherin was significantly decreased,and the protein expression levels of N-cadherin and Vimentin were significantly increased.On the contrary,after WDFY3-AS2 was overexpressed,the proliferation efficacy,immigration and invasion ability of HNSCC cell lines were significantly declined,and the protein expression levels of N-cadherin and Vimentin were significantly decreased,additionally,the protein expression level of E-cadherin was significantly increased.Conclusion(s)The prognosis risk model for HNSCC established by cuproptosis-related lncRNAs showed highly prognostic efficacy,and could be used as a molecular biomarker for diagnosis,treatment and prognosis of HNSCC,so as to provide new insights for the treatment and prognosis of HNSCC.