| Background Esophageal Carcinoma(EC)is a kind of malignant tumor that threatens human health and life.China is one of the countries with high incidence and mortality in the world.Squamous cell carcinoma is the most common histologic type of esophageal Carcinoma.The prognosis of early esophageal carcinoma patients is fine but the prognosis of advanced patients with surgery,radiotherapy and chemotherapy is very poor.Because of the high incidence and mortality,its etiology has become a research hot spot.Chromosomal helicase/ATPase DNA binding protein 1-like(CHD1L)is a newly discovered oncogene located in chr1q21,which is amplified in many solid tumors.CHD1 L is a member of the SNF-2 family.It has a highly conservative function of helicase,which regulates transcription,reconstitution chromatin and regulates the interaction between DNA and protein.However,the expression of CHD1 L protein and its clinical significance in esophageal carcinoma are uncertain,let alone the lack of the study of the biological function in the esophageal carcinoma cell lines.Objective(1)To detect the expression of CHD1 L in different esophageal carcinoma tissues by immunohistochemistry;To explore the correlation with the occurrence,development and prognosis;To provide a theoretical basis for the evaluation of clinical diagnosis,treatment and prognosis.(2)To detect the expression protein level of CHD1 L in different human esophageal carcinoma cell lines was by Westernblot,so as to prepare for the follow-up biological function research.(3)To explore the impact of CHD1 L on esophageal carcinoma cell migration,proliferation apoptosis and other biological function of CHD1 L gene by knocking down endogenous CHD1 L expression through siRNA transfection;To provide experimental evidence for the biological treatment of human esophageal carcinoma.Material and methods(1)Immunohistochemistry: To detect the expression of CHD1 L in the cancer tissues of patients with different esophageal carcinoma through paraffin section and immunohistochemistry-SP.The experimental subjects were the paraffin blocks of 191 patients with esophageal carcinoma treated by thoracic surgery in the cancer hospital of Fujian province.(2)SPSS 22 statistical software was used to analyze the patients' clinicopathological and follow-up data.Kaplan-Meier method(Log-rank test),univariate and COX multivariate regression models were used to analyze survival of patients with esophageal carcinoma,so as to provide theoretical basis for clinical diagnosis,treatment and prognosis evaluation.(3)Western blot: the expression of protein level of CHD1 L in different esophageal carcinoma cell lines was analyzed and according to the results,we selected the cell lines with high expression of CHD1 L gene.The esophageal carcinoma cell lines were KYSE150 and EC9706.To prepare for further study of its biological function.(4)A siRNA was transfected into the human esophageal carcinoma cell line EC9706.The biological functions of CHD1 L gene in migration,proliferation and apoptosis of human esophageal carcinoma cells were preliminarily studied by wound healing assays,CCK-8 cell proliferation and flow cytometry.Results The immunohistochemical results showed that the expression of CHD1 L in different esophageal carcinoma tissues was different,and its expression increased with the increase of tumor malignancy(P<0.05).The expression of CHD1 L was related to differentiation,clinical stage and metastasis of lymph nodes(P<0.01).There was no correlation between the expression of CHD1 L and the gender,age,size of tumor,location of tumor and gross types(P>0.05),those pairs of above have no significant difference.(1)Univariate analysis showed that the high expression of CHD1 L protein had a very significant effect on the overall survival rate(including 3 year survival rate and 5 year survival rate)of esophageal carcinoma patients(P<0.01).The clinical stage,differentiation degree and lymph node metastasis of esophageal carcinoma have significant impact on the prognosis of patients(P<0.01);The age,gender,gross types,size of tumor and location of tumor have no significant impact on the prognosis of esophageal carcinoma(P>0.05).(2)Multivariate Cox analysis of prognostic factors showed that tumor differentiation,pathological stages and lymph node metastasis are risk factors for survival in patients with esophageal carcinoma(P<0.05).This suggests that they can serve as a reliable indicator for prognosis of esophageal carcinoma.(3)Westernblot results showed that the expression of endogenous CHD1 L gene in EC9706 cell line was significantly higher than that in KYSE150 cell line(P<0.05),and the difference was statistically significant.(4)siRNA transfection assay showed that CHD1L-siRNA#4(5 'ACAAACTCTTGCAGCCATT-3')sequence could effectively knock down the expression level of endogenous CHD1 L protein(P<0.01),and the difference is statistically significant.(5)The results of wound healing assay test showed that knocking down of CHD1 L can effectively reduce the migration ability of esophageal carcinoma cell line(P<0.01)compared with the group without CHD1 L knockdown,and the difference was statistically significant.(6)The results of cell proliferation assay showed that knocking down of CHD1 L could effectively reduce the proliferation ability of esophageal carcinoma cells(P<0.01)compared with the group without CHD1 L knockdown and blank group.The difference was statistically significant.(7)The results of apoptosis assay showed that knocking down CHD1 L significantly increased the apoptosis level of esophageal cancer cells(P<0.01)compared with the group without CHD1 L knockdown.The difference was statistically significant.Conclusion(1)The expression of CHD1 L is associated with esophageal cancer: with the rising level of CHD1 L expression,the tumor malignancy also increases;The expression of CHD1 L has no statistically significant to the gender,age,tumor size,tumor location,and gross types.(2)The high expression of CHD1 L,clinical stage,differentiation and lymph node metastasis are independent risk factors that affect the prognosis of esophageal carcinoma,which can be used as reliable indicators.(3)The prognosis of patients with esophageal carcinoma has no statistically significant with age,sex,tumor size,tumor size and tumor location.(4)The expression of CHD1 L protein in human esophageal carcinoma cell line EC9706 is significantly higher than that of KYSE150 cell line.The EC9706 cell line can be used as a good material for further research.(5)The overexpression of CHD1 L in human esophageal cancer cells can promote the migration and proliferation ability of tumor cells and inhibit the apoptosis level,which provides a meaningful experimental basis for finding biological therapy for human esophageal carcinoma. |
PDF Full Text Request