The Expression Of WAVE3 In Esophageal Squamous Cell Carcinoma And Its Effect On Cell Ability Of Migration And Invasion

Esophageal cancer is a common malignant tumor of the digestive system,ranking fourth in the world for malignant tumors.The incidence of esophageal cancer in China ranks fifth of the all malignant tumors,and the mortality rate ranks fourth.Although with the development of medical diagnosis and treatment technology,the current treatment of the disease has made great progress,but the five-year survival rate of patients is still relatively low,the main reason is that the early symptoms of esophageal cancer are not typical,the patient had a late stage of cancer progression and lost the opportunity for radical surgery.In addition,most of the patients had postoperative metastasis and recurrence.It can be seen that early detection of tumors and reduction of tumor metastasis are key issues for improving the survival rate of esophageal cancer.The WASP family(Wiskott-Aldrich syndrome protein family,WASP family)is a novel actin regulatory protein.It has been found that its deletion causes cells to exhibit abnormalities in structure,migration and invasion.As one of the members of WASP,WAVE3 plays an important role in the migration and invasion of tumor cells.ObjectiveTo detect the expression of WAVE3 in cancer tissues of patients with esophageal squamous cell carcinoma,and to investigate the significance of WAVE3 expression in the development of esophageal squamous cell carcinoma,and to further test its expression in the serum of patients with tumor,combined with in vitro experiments.To study its effect on the migration ability of esophageal cancer cells,and provide new ideas for early diagnosis,clinical stage and treatment of esophageal squamous cell carcinoma.Methods1.Immunohistochemical method to detect the expression of WAVE3 in esophageal squamous cell carcinomaImmunohistochemical method was used to detect the expression of WAVE3 in 62 cases of esophageal squamous cell carcinoma and corresponding adjacent tissues.Combined with relevant clinical medical records,the expression of WAVE3 was correlated with the occurrence and development of esophageal squamous cell carcinoma.2.Real-time PCR method for detecting the expression levels of WAVE3 mRNA and miRNA200 b in serum of patients with esophageal squamous cell carcinoma and normal subjectsReal-time PCR was used to detect the relative levels of WAVE3 mRNA and miRNA200 b in serum of 80 patients with esophageal squamous cell carcinoma and 30 healthy subjects.The expression levels of WAVE3 mRNA and miRNA200 b and the correlation between the two groups were compared.The clinical data were used to analyze whether the expression level of WAVE3 mRNA in serum was correlated with the tumor stage and metastasis of esophageal squamous cell carcinoma.3.Esophageal squamous carcinoma cell line EC109 was cultured in vitro to explore the effect of WAVE3 on its migration ability1)Esophageal squamous cell carcinoma cell line EC109 was cultured in vitro,and WAVE3 expression was inhibited by transfection of miRNA200 b.The transfection efficiency was detected by real-time PCR and the inhibition rate of WAVE3 protein Objective expression was detected by Western Blot.2)Transwell assay detects the changes of EC109 migration ability after inhibition of WAVE3 protein expression.Results1.The results of immunohistochemistry showed that the positive expression rate of WAVE3 protein in esophageal squamous cell carcinoma(20.9%)was higher than that in adjacent tissues(4.8%),and the difference was statistically significant(χ2=11.317,P <0.05).Analysis with clinicopathological data,the positive rate of WAVE3 expression was correlated with tumor TNM stage,tumor invasion depth and metastasis.The positive rate of TNM stage III/IVWAVE3 expression(36.4%)was higher than that of TNM stage I/II(12.5).%),the positive rate of WAVE3 expression in the deep infiltration/deep layer of the tumor(24%)was higher than that in the submucosa/superficial layer(8.3%),and the positive rate of WAVE3 expression in patients with lymph node metastasis(33.3)%)was higher than patients who did not have metastases(14.6%).2.The results of real-time PCR showed that the level of WAVE3 mRNA in serum of the patients with esophageal squamous cell carcinoma was higher than that of normal subjects(5.325(4.683,7.123)vs 3.180(2.130,4.020)),while the expression level of miRNA200 b was lower than that of normal subjects(2.440(1.880,3.002)vs 3.001(2.130,4.100)),the difference was statistically significant,and the expression level of WAVE3 mRNA was negatively correlated with the expression of miRNA200 b.Analysis with clinical pathological data,the expression level of serum WAVE3 mRNA in esophageal squamous cell carcinoma patients was associated with TNM staging and lymph node metastasis of esophageal carcinoma.The high expression was expressed in patients with stage TNM stage III/IV and lymph node metastasis.3.The results of real-time PCR showed that the expression level of miRNA200 b in esophageal squamous cell carcinoma cell line EC109 transfected with miRNA200 b positive group was higher than the blank control group and the negative control group(19.065±8.980 vs 1.000±0.000,0.964±0.997).Subsequent WB results showed that the level of WAVE3 protein in the positive experimental group decreased after the expression of miRNA200 b was increased.The results of Transwell assay showed that the number of cell migration(51.33±6.03)in the positive experimental group of EC109 was lower than the blank control group(114.67±4.73)and the negative control group(117.00±3.61).Conclusions1.The expression of WAVE3 protein is elevated in esophageal squamous cell carcinoma,and its high expression may play a crucial role in the progression of ESCC and is associated with poor prognosis.2.The expression of WAVE3 protein in serum of patients with ESCC may be associated with decreased expression of miRNA200 b,and its high expression is associated with the course of esophageal squamous cell carcinoma,and is expected to become a new biological marker.3.Exogenous miRNA200 b can inhibit the expression of WAVE3 protein in esophageal squamous carcinoma cells,thereby inhibiting the migration ability of cancer cells,and providing new therapeutic targets and strategies for the treatment of esophageal squamous cell carcinoma.