Correlation Research Of CCL1 Expression In Cervical Cancer And Treg Cells Infiltration And The Clinical Pathological Characters

Objective:To observe the expression of chemokine CCL1 in cervical carcinoma cells and analyze its correlation with the number of CCR8+FOXP3+Treg cells,and to investigate the significance of CCL1-CCR8 in promoting the infiltration of Treg cells; To analyze the correlation of CCL1 expression with clinical pathological characters; to detect the level of CCL1 gene expression in different subtypes of HPV cervical cancer cell lines using real-time PCR.Methods:The expression of chemokine CCL1 in cervical carcinoma, HSIL, and normal cervical tissue was detected by immunohistochemistry. Immunofluorescent double-staining was used to detect the expression of chemokine receptor CCR8 in FOXP3+ Treg cells in cervical carcinoma tissue. Statistical analysis was used to determine the correlation between CCL1 expression in cancer cells and the number of infiltrating CCR8+FOXP3+ Treg cells in the stroma of cervical cancer. The relationship between CCL1 expression of cervical cancer with the depth of invasion, metastasis of lymph nodes, the degree of differentiation, the type of pathology, and the age of patients was analyzed. Culture HPV-positive(Hela cell, SiHa cell) and HPV-negative(C33A cell)cervical carcinoma cell line in vitro, and real-time PCR was used to detect and compare the differences of the expression of CCL1 gene between HPV-positive and HPV-negative cervical cancer cell line.Results:Normal cervical tissue did not express chemokine CCL1; a small part of HSIL weakly expressed CCL1, and the positive rate was 10%(2/20); the positive rate of CCL1 in cervical carcinoma was 45%(27/60), which showed significant difference compared with normal cervical tissue and HSIL(p<0.01,p<0.01). Immunofluorescent double-staining showed that FOXP3+T cells expressed CCR8. In the CCL1+ cervical cancer group, the number of infiltrating CCR8+FOXP3+Treg cells was significantly greater than that in the CCL1- cervical cancer group. The expression of CCL1 was positively correlated with the number of infiltrating FOXP3+ Treg cells(p<0.01). Statistical correlation analysis showed that the CCL1 expression of the cancer was closely related to their lymph node metastasis(p<0.05); however, it was not associated with depth of invasion, the degree of differentiation, the type of pathology, and the age of patients. Real-time PCR showed that the level of CCL1 gene expression in HPV-16 positive(SiHa)and HPV-negative cell line(C33A) was greatly higher than that in HPV-18 positive cell line( Hela)(p<0.01,p<0.05).Conclusion:1. The expression of CCL1 in cervical carcinoma was greater than that in normal cervical tissue and HSIL.2. The expression of CCL1 was associated with the increasing number of infiltrating FOXP3+CCR8+T cells, which showed that CCL1-CCR8 axis might play a role in the promotion of Treg infiltration and inhibition of antitumor immunity. CCL1-CCR8 may become a new target for the therapy of cervical cancer.3. The expression of CCL1 in cervical carcinoma cells was related to lymph node metastasis of cervical cancer.4. The level of CCL1 gene expression in cervical cancer cell lines was: SiHa> C33A>Hela, and HPV16 positive cell line may be screened as a cell line which highly expressed CCL1 gene in the next phase of the experiment.