Expression Of The C5aR1 In Esophageal Squamous Cell Carcinoma And Preliminary Study On The Molecular Mechanisms Of C5aR1 Impact Esophagus Carcinogenesis On The Malignant Biological

Objective: Esophageal cancer(EC)is one of the most familiar malignant neoplasms that seriously threaten human health and life.China has the largest number of new cases of esophageal cancer in the world,and there are more than half of the esophageal cancer patients in the world.Esophageal squamous cell carcinoma(ESCC)accounts for more than 90% of all esophageal cancer cases in China.Although there are many treatments for esophageal cancer,the survival prognosis is still poor.Therefore,it is essential to investigate new biological targets.C5aR1(C5a allertoxin chemoattractant receptor,C5a-R or CD88)is a classical G protein-coupled receptor that plays a headstone function in the inherent immune response.Recent studies have discovered that the expression level of C5aR1 is increased in various tumors,which plays a role in promoting the occurrence and progression of tumors.However,the expression of C5aR1 in ESCC and whether C5aR1 has a cancer-promoting effect and the possible molecular mechanism remain unclear.The aim of this study is to investigate the expression of C5aR1 in ESCC and its relationship with clinicopathological parameters and prognosis,and to explore the effect and mechanism of C5aR1 on the biological behavior of human ESCC cell lines KYSE410 and KYSE150 in vitro.Method: GEPIA database was used for bioinformatics analysis of C5aR1expression in ESCC and control tissues and its relationship with prognosis.The expression of C5aR1 protein in ESCC and para-carcinoma tissues was detected by immunohistochemistry.The relationship between C5aR1 protein expression and clinicopathological parameters were analyzed,and the effect of C5AR1 protein expression on the prognosis of patients with ESCC was analyzed.Western Blot was used to detect the expression of C5aR1 protein in KYSE410 and KYSE150 cells.Transient transfection was used to construct C5aR1knockdown group and control group in KYSE410 and KYSE150 cell lines,respectively.q RT-PCR and Western Blot were used to verify the interference efficiency.CCK8 assay,colony formation assay,wound healing assay and Transwell assay were used to detect the effect of C5aR1 knockdown on the biological behavior of esophageal cancer cells.EMT protein detection: Western Blot was used to detect E-cadherin and Vimentin protein in esophageal cancer cell line C5aR1 knockdown group and control group.Exploration of possible molecular mechanism: Western Blot was used to detect the expression of nuclear protein β-catenin in C5aR1 knockdown group and control group.Result: The results of bioinformatics analysis showed that C5aR1 was highly expressed in esophageal cancer,and a high expression of C5aR1 was connected with a poor prognosis.The results of immunohistochemistry showed that the expression of C5aR1 protein was high in ESCC tissues and low in paracarcinoma tissues(c2=12.481,P < 0.001).C5aR1 protein was associated to the grade of delineation(c 2=7.066,P=0.029)and the recurrence of tumors(c 2=3.998,P=0.046).The intensity and range of C5aR1 protein expression were stronger and wider in high-grade tumors and recurrent cases.However,there was no suggestively association with age,gender,tumor size,lymph node metastasis and T stage(P >0.05).Survival analysis presented that the overall survival time of C5aR1 high expression group was worse than that of C5aR1low expression group(c2=4.066,P=0.044),and C5aR1 was a factor affecting the survival of patients.The impact of C5aR1 knockdown on the tumor biology of esophageal cancer cells in vitro: the proliferation,migration and invasion ability of C5aR1 knockdown group were weakened when compared to the control group in vitro.Impact of C5aR1 knockdown on EMT: compared with the control group,the quantity of E-cadherin protein was up-regulated and Vimentin protein was down-regulated in the C5aR1 knockdown group.Potential mechanism of action: the expression of β-catenin in nuclear protein was down-regulated in C5aR1 knockdown group.Conclusion : C5aR1 may be involved in the occurrence and development of ESCC,and high expression of C5aR1 protein indicates poor prognosis of patients with ESCC.Knockdown of C5aR1 impaired the proliferation,migration and invasion of esophageal cancer cells in vitro.Knockdown of C5aR1 inhibited the EMT process of esophageal cancer.C5aR1 may affect the progression of esophageal cancer by regulating β-catenin.