Histone Lactylation Affects Tumorigenesis By Regulating TRIM29 Expression In Lung Squamous Carcinoma

Purpose:Lung cancer is the most common malignant tumor in China in terms of incidence and mortality.Among them,Lung squamous cell carcinoma(LUSC)is the second most common tissue type of lung cancer.The major driver gene of LUSC has not been identified,and there is still no molecularly targeted drug with definite clinical efficacy.Previous studies have demonstrated that TRIM29 is an oncogene in lung squamous carcinoma,but studies of TRIM29 in lung squamous carcinoma are lacking and the upstream regulatory mechanism of TRIM29 is unclear.Our group used bioinformatics,cell biology,and molecular biology to investigate the mechanism of histone lactonization through TRIM29 to regulate the biological behavior of lung squamous carcinoma cells.This study provides a new potential target for molecularly targeted therapy of lung squamous carcinoma.Methods:1.Bioinformatics,clinical lung squamous carcinoma paraffin-embedded tissue sections to analyze the expression of TRIM29 in cancer and paracancerous normal tissues,and analyze the clinical characteristics and prognostic relevance of TRIM29 to patients with lung squamous carcinoma.2.RT-qPCR,Western blot,and cellular immunofluorescence were used to verify the expression of TRIM29 in lung squamous carcinoma cell lines and the effect of siRNA transfection targeting TRIM29.CCK8 proliferation assay,cell scratching,and other experiments confirmed the effect of TRIM29 expression changes on the biological behavior of lung squamous carcinoma cells.3.Bioinformatics analysis of LDHA expression in cancer and normal tissues adjacent to cancer in lung squamous carcinoma;immunohistochemical analysis of cellular expression localization of histone lactonized Pan Kla and H3K181a in paraffin-embedded tissues of lung squamous carcinoma;RT-qPCR,Western blot,and cellular immunofluorescence to verify the expression of Pan Kla and H3K181a in lung squamous carcinoma cell lines.4.CCK8 proliferation assay,cell scratching,cell migration and invasion assay,clone formation assay to analyze the effect of glycolysis inhibitors on the biological behavior of lung squamous carcinoma cells;Western blot,RT-qPCR to verify the changes of histone lactonized Pan Kla,H3K181a levels and TRIM29 expression in lung squamous carcinoma cell lines after the intervention of glycolysis inhibitors.Results:1.By downloading and analyzing the gene expression and clinicopathological information of lung squamous carcinoma from the TCGA database,the results showed that TRIM29 was highly expressed in lung squamous carcinoma compared with the corresponding normal lung tissue(P<0.05);TRIM29 expression did not correlate with clinical characteristics and survival time of patients;immunohistochemical results showed that TRIM29 was localized in the cytoplasm.2.TRIM29 expression was higher in both lung squamous carcinoma cell lines NCI-H520 and NCI-H226 compared with normal bronchial epithelial cells BEAS-2B;TRIM29 siRNA could effectively down-regulate the expression level of TRIM29 protein;the proliferation and migration ability of lung squamous carcinoma cells decreased after down-regulating TRIM29 expression.3.By downloading and analyzing the gene expression and clinicopathological information of lung squamous carcinoma from the TCGA database,the results showed that LDHA was highly expressed in lung squamous carcinoma compared with the corresponding normal lung tissue;immunohistochemistry showed that both Pan Kla and H3K181a were localized in the nucleus;compared with normal bronchial epithelial cells BEAS-2B,lung squamous carcinoma cell lines NCI-H520 and NCI Histone lactonized Pan Kla and H3K181a levels were elevated in H226 compared with normal bronchial epithelial cells.4.The results of CCK8 proliferation assay,cell scratching,cell migration and invasion assay,and clone formation assay showed that glycolysis inhibitors could effectively inhibit the proliferation,migration,and clone formation ability of lung squamous carcinoma cells;and glycolysis inhibitors could inhibit histone lactylation Pan Kla and H3K181a modification,and the mRNA and protein expression levels of TRIM29 were decreased.Conclusion:1.TRIM29 is an important pro-oncogene in lung squamous carcinoma and can be used as an important biomarker for the diagnosis of lung squamous carcinoma disease.2.Reduced levels of histone lactylation inhibit the progression of lung squamous carcinoma.3.Histone lactylation regulates the expression of TRIM29 to influence lung squamous carcinoma progression.