Study On The Function Of FZD6 In Esophageal Squamous Carcinoma

Objective 1 Exploring the expression specificity of Frizzled6(FZD6)in esophageal squamous cell carcinoma.2 Exploring the influence of FZD6 on the migration,invasion,proliferation,cloning and other phenotypes of esophageal squamous cell carcinoma.3Exploring whether FZD6 is related to transcription factors TP63 and SOX2.Methods According to public database analysis,compared with normal tissues,FZD6 expression in a variety of tumors,in the specific histological classification of esophageal cancer,whether FZD6 expression differences are biased.Collect matched tissue samples from 64 patients with esophageal cancer(samples are from Cancer Hospital of Chinese Academy of Medical Sciences and Beijing 301 Hospital),and the tissue samples were subjected to TRLzol grinding to extract total RNA,and RT-q PCR experiment was performed to detect FZD6 expression in cancer and paracancerous tissues Analyze the situation.Esophageal squamous cell carcinoma and normal cell lines were used to extract total RNA with TRLzol lysate,and RT-q PCR experiments were performed to analyze the difference in FZD6 expression in the cell lines.The immunohistochemical staining experiment was used to stain the esophageal cancer tissue chip to analyze the expression of FZD6.The RNAi method was used to perform knockdown experiments on cell lines.Plasmid transfection was used to perform overexpression experiments on the cell line.The efficiency of knockdown and overexpression was verified by RT-q PCR experiment and Western bloting.The CCK-8 experimental method was used to detect the effect of overexpression of FZD6 on cell proliferation.Transwell experiment was used to detect the effect of FZD6 overexpression and knockdown on the invasion and migration ability of esophageal squamous cell carcinoma cells.The plate cloning experiment was used to detect the effect of knockdown of FZD6 on the cloning ability of esophageal squamous cell carcinoma.The RNAi method was used to knock down the transcription factors TP63 and SOX2 in the esophageal squamous cell carcinoma cell line,and the RT-q PCR experiment was used to detect the knockdown efficiency of transcription factors and the changes of FZD6 after knockdown.Plasmid transfection was used to perform overexpression experiments on TP63 and SOX2 in the cell line,and RT-q PCR experiments were used to detect the overexpression efficiency of transcription factors and the changes of FZD6 after overexpression.Results 1 Database analysis showed that the expression of FZD6 in esophageal cancer was significantly higher than that in normal tissues,and the expression of FZD6 in esophageal squamous cell carcinoma was significantly higher than that in esophageal adenocarcinoma and normal tissues,while the expression of FZD6 in esophageal adenocarcinoma was similar to that in normal tissues.The expression level of the difference is not obvious.2Immunohistochemical analysis showed that the expression of FZD6 in cancer tissues was significantly higher than that in adjacent tissues.3 In the RT-q PCR experiment,FZD6 expression in esophageal squamous cell carcinoma cell lines was significantly higher than that of normal esophageal epithelium.4 After RNAi interference,the expression of FZD6 was significantly lower than that of the control group,and the expression of FZD6 after plasmid transfection was significantly higher than that of the control group.5 In the CCK8 experiment,the cell proliferation of the FZD6 knockdown group was significantly lower than that of the control group.6 In the Transwell experiment,the invasion and migration ability of the cells in the FZD6 knockdown group was significantly lower than that of the control group,and the invasion and migration ability of the cells in the FZD6 overexpression group was significantly higher than that of the control group.7 In the plate cloning experiment,the cloning ability of the cells in the FZD6 knockdown group was significantly lower than that of the control group.8 After the RNAi method interfered with the cell line,the expression levels of TP63 and SOX2 were significantly lower than those of the control group,and the expression levels of TP63 and SOX2 were significantly higher than the control group after plasmid transfection.9 After knockdown of TP63 and SOX2,the expression of FZD6 was lower than that of the control group.After overexpression of TP63 and SOX2,the expression of FZD6 was higher than that of the control group.Conclusion 1 FZD6 is specifically and highly expressed in esophageal squamous cell carcinoma.2 FZD6 can promote the invasion,migration,proliferation and cloning ability of esophageal squamous cell carcinoma.3 The expression of FZD6 is regulated by transcription factors TP63 and SOX2.Figure 15;Table 1;Reference 149...