The Expression And Application Of Human Leucocyte Antigen-G (HLA-G) In Cervical Cancer And Intraepithelial Neoplasia

Objective:To detect the expression of the membrane-bound HLA-G in the tissue or the solube HLA-G in the serum of patients with cervical cancers compared with normal controls. Detect the expression of IL-10 in the serum of the patients with cervical cancer. Explore its effect and significance in the development of cervical cancer. Discuss its possible clinical application in diagnosis of cervical cancer compared with SCC or CA125.Methods:1. Immunohistochemistry S-P method was used to detect the expression of membrane-bound HLA-G Immunohistochemistry S-P method was used to detect the expression of membrane-bound HLA-G in 54 cases of cervical cancer tissues and 17 cases of normal cervix tissues, and its relationship to clinical pathological characteristics of cervical cancer was also analyzed.2. Western blot method was used to confirm the expression of membrane-bound HLA-G protein in cervical cancer 9 specimens of patients with cervical cancer and 7 specimens form non-tumorous controls were detected using Western blot method to confirm the expression of membrane-bound HLA-G protein and to illustrate the difference from normal controls.3. Using enzyme-labeled immunosorbent assay (ELISA) method to detect soluble HLA-G protein levels ELISA was used to detect plasma of 152 cervical lesions (including 80 cervical cancer,15 CINⅠ,22 CINⅡand 35 CINⅢ) and 20 normal controls, compare the differences among them and analyze its possible role in tumor formation.4. Using ELISA to detect IL-10, SCC and CA125 levels in cervical cancer Through detection of IL-10, SCC and CA125 levels in plasma of cervical lesions and normal controls by ELISA, to initially explore the mechanism that HLA-G help tumor cells escape from host immune system.5. Using ROC curve to analyze the early-stage diagnosis values of sHLA-G, SCC and CA125, respectively. Compared all results, to evaluate the possible diagnosis values of sHLA-G in lesions before formation of cervical carcinomaResult:1. The positive expression rate of HLA-G was 66.7% (36/54) in cervical cancer, while no positive expression (0/17) was found in normal cervix tissues (P＜0.05). The positive expression rate of HLA-G was closely related with clinical stage and differentiation degree (P＜0.05), but no relationship to age, histological classification, and lymph node metastasis (P＞0.05)2. To confirm the existence of HLA-G in human cervical carcinoma, we also detected the HLA-G protein by Western blot. Evidence for HLA-G antigens, indicated by the occurrence of a discrete band with a molecular weight of about 39kDa, was detected in HLA-G positive and in none of adjacent tumor-negative cervical tissues.2. Soluble HLA-G levels in the carcinoma group(191.4U/ml), CIN3 group(109.60U/ml), CIN2 group(89.69U/ml) and CIN1 group(46.24U/ml) were significantly higher than normal controls(45.18U/ml) (P＜0.05). These results suggested the important role of HLA-G in immune escape. Soluble HLA-G levels were significantly correlation with differentiation parametrial invasion and lymph-node metastasis, suggested that sHLA-G may have asosiation with development of cervical carcinoma and be used as a prognosis factor. 3. The mean serum IL-10 levels were 3.10pg/ml in the normal group,9.16 pg/ml in the CIN1 group,15.09pg/ml in the CIN2 group,17.04pg/ml in the CIN3 group, and 26.45pg/ml in the carcinoma group. The results could established a poor positive correlation (r= 0.311, P＜0.05) of IL-10 and sHLA-G levels, which indicated that IL-10 could up-regulate the expression of HLA-G, thus correlation between the two was evaluated in the current study.4. Analysis of ROC curve demonstrated that sHLA-G (AUC:0.710) significantly superior performance to SCC (0.643) and CA 125(0.588) (P＜0.05). The ROC analysis revealed that sHLA-G level at 108.20U/ml was the clear cut-off value for which the ability to distinguish cervical carcinoma patients from nontumorous controls was maximized. With this cut-off level, the sensitivity and specificity of sHLA-G for cervical carcinoma were 73.30% and 65.71%, respectively. From the results, sHLA-G, as a diagnosis indicator in early-stage, had better values than SCC and CA125.Conclusion:1. Expression of HLA-G in cervical tumors is significantly higher than that in normal tissues, and HLA-G also plays an important role in the development of cervical cancer.2. A positive correlation is established between soluble HLA-G expression and levels of IL-10, which indicates they may enhancement down-regulation immune function and promote the growth of tumour cells.3. Compared with SCC and CA125, sHLA-G has ascendancy in the diagnosis of early-stage in cervical cancer, and can investigate further to clinical application.