| Objects: Objective to screen and identify the key genes that can regulate the drug resistance and tumor immunity of head and neck squamous cell carcinoma(HNSCC).Methods: Cisplatin resistant Fa Du cells were screened by drug sensitivity test combined with CRISPR/Cas9 library technology,and then DNA was extracted and sequenced to obtain cisplatin resistant genes.Based on the bioinformatics methods,we downloaded the m RNA data and clinical data of HNSCC samples from the TCGA database to screen for immune-related different expression genes(IRDEGs)in HNSCC patients.Univariate and multivariate Cox regression analysis,Kaplan Meier survival analysis and ROC curve analysis were performed on these data.Least Absolute Shrinkage And Selection Operator(LASSO)was used to further screen and establish a risk score model that can predict the survival and prognosis of HNSCC patients,and GSE65858 HNSCC sample data was used to verify this model we established.Finally,IRDEGs and cisplatin resistance related genes that can simultaneously regulate the survival and prognosis of HNSCC patients were screened by using Veen diagram,and the preliminary functional verification was carried out.Results:1.The lowest lethal concentration of cisplatin in Fa Du cells was 30μM.2.Through CRISPR / Cas9 library technology,11504 key genes that may be involved in Fa Du cisplatin resistance were screened.3.There are 31 survival related IRDEGs in HNSCC samples,among which 9 key prognostic factors,including IGSF5,NKX2-3,HLF,ALDH2,IFIT2,FXYD5,CTSL1,IFNAR1 and RNF216,can jointly construct a risk score model for survival and prognosis of HNSCC patients.4.In univariate cox regression analysis,clinical characteristics such as N stage,T stage,stage,age and constructed prognostic model were significantly correlated with survival.In multivariate cox regression analysis,N stage,age and constructed prognostic model were independent prognostic factors.According to the risk score model,the prognosis of the high-risk group was worse than that of the low-risk group,the immune score of the high-risk group was significantly higher than that of the low-risk group,and the proportion of cells naive,plasma cells,T cells CD4 memory resting and other 10 kinds of cells in the high-risk and low-risk samples was significantly different(P < 0.05).5.IGSF5 is an immune-related gene with significantly low expression in HNSCC,which may be involved in the regulation of tumor chemotherapy resistance.Overexpression of IGSF5 can inhibit the proliferation of HNSCC.Conclusions: The cisplatin resistance related genes screened by CRISPR/Cas9 system have high reliability.IRDEGs such as IGSF5,which are related to the prognosis of HNSCC,can be used as a prognostic risk assessment model for HNSCC,which is helpful to systematically assess the survival and prognosis of HNSCC patients.IGSF5,an immune-related gene significantly low expressed in HNSCC,may be involved in regulating the cisplatin resistance of HNSCC. |
