Study On The Correlation Between FZD Family And Lung Adenocarcinoma

Lung cancer is one of the most common malignancies worldwide.The histological classification of lung cancer can be divided into squamous cell carcinoma,adenocarcinoma,adenosquamous carcinoma,small cell carcinoma,large cell carcinoma and sarcomatoid carcinoma.The incidence of lung adenocarcinoma is second only to squamous cell carcinoma,and its incidence has an obvious increasing trend.In some areas,the incidence of lung adenocarcinoma and squamous cell carcinoma are almost equal,and female patients with lung adenocarcinoma are relatively common,accounting for more than half.The clinical treatment effect and prognosis of lung adenocarcinoma is inferior to that of squamous cell carcinoma,and the survival rate is less than 10% 5 years after surgical resection.In recent years,immunotherapy and molecular targeted therapy of lung adenocarcinoma have gradually become a research hotspot.In the study of lung adenocarcinoma,bioinformatics database data were used to screen and analyze genes related to the occurrence and development of lung adenocarcinoma,to search for potential therapeutic target genes,and drugs were used to block relevant signal transduction pathways,thus affecting the growth and apoptosis of tumor cells.The Wnt signaling pathway is involved in the development of a variety of human tumors,including lung cancer,liver cancer,colorectal cancer,ovarian cancer,breast cancer,prostate cancer,osteosarcoma,and nephroblast.Wnt signaling pathways are mainly composed of 4 pathways.Wnt cell plane polarization pathway,intracellular pathway regulating asymmetrical cell division in spindle direction,Wnt/Ca2+ pathway,Wnt / β-catenin pathway(classical Wnt signaling pathway).Wnt / β-catenin signaling pathway is an open signaling pathway with multiple links and multiple action sites.After activation,Wnt ligand combines with Frizzled Fzd to transmit signals from the cell surface to the nucleus,affecting the growth,proliferation,metastasis and differentiation of cells,and related to the proliferation and metastasis of tumors and other biological functions.The FZD(Frizzled)protein family is the transmembrane receptor of the Wnt signaling pathway.It is found that the Frizzled protein family includes 10 Frizzled proteins and one Smoothened protein in mammals.The motif composition of different proteins is different,but all contain a conserved KTXXXW motif at the C end.Current studies have found that members of the FZD family are associated with a variety of human tumors,such as increased FZD10 expression in gastric cancer tissue,positive correlation between FZD5 expression and malignant degree of glioma,and FZD3 may play a role as a potential oncogene in the occurrence and development of colorectal cancer.Carrera W et al.found that mutations in the FZD4 gene were associated with familial exudative vitreoretinopathy.In this study,the gene expression of FZD family genes(FZD1-FZD10)in normal tissues and lung adenocarcinoma tissues was analyzed sequentially by bioassay,and the genes related to the family and lung adenocarcinoma were searched for.To study the gene expression of FZD family genes in TNM stage and clinical stage of lung adenocarcinoma.The relationship between FZD family gene expression and the survival time of lung adenocarcinoma;Relationship between FZD family gene expression level and tumor microenvironment and immune cells in lung adenocarcinoma.Finally,cell biology experiments were conducted to detect the effect of high expression of related genes in the FZD family on lung adenocarcinoma cells(NCI-H1299 cells)through cell transfection technology,quantitative real-time fluorescence PCR(qRT-PCR),scratch test,Transwell invasion test,flow cytometry and other methods.Methods:1.The expression of FZD family genes in normal samples and lung adenocarcinoma samples were analyzed by GEO,TCGA and TCGA+ GTEX database data to find the differentially expressed family members.The gene expression of FZD family genes in TNM stage and clinical stage of lung adenocarcinoma were analyzed.The expression of FZD4 gene was significantly decreased in lung adenocarcinoma tissues.2.Kaplan-Meier was used to analyze the relationship between FZD4 gene expression level and survival time of lung adenocarcinoma.Estimate was used to obtain the immune and matrix scores of lung adenocarcinoma,and the expression level of FZD4 was analyzed and compared.The score data of six types of immunoinfiltrating cells in lung adenocarcinoma were downloaded from the TIMER database,and the correlation between FZD4 gene expression and immune cells was analyzed by R language packet.3.Lentivirus transfection technique was used to high express FZD4 gene in human lung adenocarcinoma cell NCI-H1299.Cell transfection technique,quantitative real-time fluorescence PCR(qRT-PCR)was used to test the transfection rate of FZD4 gene.Flow cytometry,scratch assay and Transwell invasion assay were used to detect the effects of high FZD4 expression on differentiation,apoptosis,migration and invasion ability of NCI-H1299 cells.Results:1.The expression of FZD4 gene in FZD family was significantly decreased by combining GEO,TCGA and TCGA+ GTEX database data.There was no difference in FZD gene expression between TNM stage and clinical stage of lung adenocarcinoma.2.Lung adenocarcinoma patients with high FZD4 expression have a longer survival time;The level of FZD4 expression is positively correlated with lung adenocarcinoma immune score and stromal score;The expression level of FZD4 is positively correlated with the infiltrating immune cells of lung adenocarcinoma B lymphocytes,CD4+ T lymphocytes,CD8+ T lymphocytes,dendritic cells,macrophages,and neutrophils.3.Compared with the control group,the ratio of cells in G1/G0 phase was increased in the FZD4 overexpression vector group,while the ratio of cells in S phase and G2/M phase was decreased;FZD4 overexpression vector could significantly induce apoptosis of lung adenocarcinoma cells;Transfected with FZD4 overexpression vector,the migration and invasion ability of lung adenocarcinoma NCI-H1299 cells were significantly reduced.Conclusion:The low expression of FZD4 gene is associated with the occurrence of lung adenocarcinoma;patients with high FZD4 gene expression in lung adenocarcinoma have a longer survival time than those with low expression;FZD4 can block lung adenocarcinoma NCI-H1299 cells in the G1/G0 phase,induce the apoptosis of NCI-H1299 cells,and reduce the migration and invasion ability of NCI-H1299 cells.FZD4 gene can be used as a tumor suppressor gene in lung adenocarcinoma.