| BackgroundLung malignant tumors endanger human health,and statistics on cancer incidence and mortality in China show that lung cancer ranks first among cancers in terms of new cases.Lung cancer is classified into two types based on clinical and pathological characteristics:small cell lung cancer and non-small cell lung cancer.Non small cell lung cancer also includes lung squamous cell carcinoma,lung adenocarcinoma,large cell lung cancer,and others.Lung squamous cell carcinoma is a prevalent histological subtype of lung cancer and a leading cause of cancer fatalities.Conventional treatment approaches include surgery,radiation,and chemotherapy.However,effectiveness for people with advanced lung cancer is insufficient.With the continuous development of molecular targeted therapy in recent years,the survival time of lung cancer patients has been extended.Molecular targeted therapy is very effective for patients with sensitive gene mutations in lung cancer.However,for patients without sensitive mutations,finding new tumor biomarkers and new therapeutic targets is crucial for the diagnosis and treatment of lung cancer.Zinc finger DHHC domain-containing protein 12(ZDHHC12)belongs to the palmitoyl transferase family,which catalyzes protein palmitoylation and is overexpressed in many malignant tumors;however,the role and clinical significance of ZDHHC12 in lung squamous cell carcinoma remain unknown.Thus yet,no studies on ZDHHC12 expression in lung squamous cell carcinoma have been reported.ObjectiveThe goal of this study is to use bioinformatics to examine the association between ZDHHC12 expression,prognosis in lung squamous cell carcinoma,as well as to explore the biological function of ZDHHC12 in lung squamous cell carcinoma through in vitro experiments.Methods1.Download the data of lung squamous cell carcinoma from the TCGA database,and through bioinformatics analysis.The expression of ZDHHC12 in lung squamous cell carcinoma,its relationship with clinical characteristics,prognosis,function,modifications in gene level,and the relationship between tumor infiltrated immune cells were explored using R.2.Quantitative real-time PCR(qPCR)was used to detect the mRNA expression level of ZDHHC12 in lung squamous cell carcinoma.3.Knock down the expression of ZDHHC12 on lung squamous cell carcinoma cell line NCI-H520 and SK-MES-1,and use qPCR to detect cell ZDHHC12 gene.Next,CCK-8 method and EdU method,scratch experiment,and transwell experiment to detect the impact of ZDHHC12 on the proliferation,migration,and invasion.Flow cytometry was applied to detect cell cycle and apoptosis.4.The experiments conducted in the study were repeated three or more times.Flowjo software was used to plot and analyze streaming data,and GraphPad Prism 9.0 software was used for statistical analysis.P<0.05 is considered statistically significant.Results1.The mRNA and protein expression levels of ZDHHC12 in lung squamous cell carcinoma are significantly higher than those in normal lung tissue.2.There is a correlation between the expression of ZDHHC12 and the age of patients with lung squamous cell carcinoma,and the mRNA expression level of ZDHHC12 in patients older than 65 years is significantly reduced.However,there is no correlation between ZDHHC12 and gender,clinical TNM stage in patients with lung squamous cell carcinoma.3.The increased expression level of ZDHHC12 indicates a poor prognosis in patients with lung squamous cell carcinoma.4.The increased expression level of ZDHHC12 is related to the cAMP signaling pathway.5.ZDHHC12 is negatively correlated with B cells,CD8+T cells,and CD4+T cells in lung squamous cell carcinoma.ZDHHC12 is negatively correlated with immune checkpoint related gene CD286.qPCR validation found that ZDHHC12 expression was increased in lung squamous cell carcinoma tissues.7.Compared with the control group,knockdown of ZDHHC12 inhibits the proliferation of lung squamous cell carcinoma cells.8.The cell cycle test results suggest that there is no significant change in cell cycle after knocking down ZDHHC12.The results of cell apoptosis test shows that knocking down ZDHHC12 had no significant effect on the apoptosis level of SK-MES-1 and NCI-H520 cells.9.Compared with the control group,knockdown of ZDHHC12 inhibits the migration,and invasion of lung squamous cell carcinoma cells.10.The expression level of ZDHHC12 mRNA in lung squamous cell carcinoma is negatively correlated with the level of ZDHHC12 methylation.The increased mRNA expression level of ZDHHC12 may be caused by a decrease in DNA methylation levels.Conclusion1.The expression of ZDHHC12 is increased in lung squamous cell carcinoma tissue,and the increased expression of ZDHHC12 is associated with poor prognosis in patients with lung squamous cell carcinoma,which has the potential to become a tumor marker for lung squamous cell carcinoma.2.In vitro experiments have verified that knockdown ZDHHC12 gene inhibits the proliferation,migration,and invasion of lung squamous cell carcinoma cell lines. |
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