An Experimental Study On Expression Of HLA-G In Ovarian Cancer Cell Line SKOV-3

Objective:To investigate the expression of human leucocyte antigen-G (HLA-G)in ovarian cancer cell line SKOV-3.Methods:The expression of HLA-G in choriocarcinoma cell lines JEG-3 as the positive control,we used Rt-PCR,immunocytochemical method and flow cytometry to detect the expression of HLA-G in ovarian cancer cell line SKOV-3.(1)When the cells of ovarian cancer cell line SKOV-3 grow to 5×10⁴/ml,collect cells,then extract RNA from cells using Trizol,detect the expression of HLA-G mRNA using Rt-PCR. (2)Inoculate the cells of ovarian cancer cell line SKOV-3 into a six-hole plate with germ-free coverslips,take out of coverslips after being cultured in a 37℃,5% CO₂-humidified incubator with 48 hours,detect the expression of HLA-G using immunocytochemical method.(3)Collect the cells of ovarian cancer cell line SKOV-3 and make them into monoplast suspension,detect the expression of HLA-G using flow cytometry.Results:(1)HLA-G were found at the membrane and the cytoplasm by immunocytochemical method.(2)HLA-G mRNA were found in ovarian cancer cell line SKOV-3 by Rt-PCR.HLA-G1:764bp HLA-G2:488bp HLA-G3:215bp HLA-G4:491bp HLA-G5:886bp HLA-G6:610bp HLA-G7:360bp(3)The results of flow cytometry showed the rate of the HLA-G-positive tumor cells is 33.77%.Conclusions:The expression of HLA-G mRNA and HLA-G protein in ovarian cancer cell line SKOV-3 were positive,HLA-G is involved in the immune escape of ovarian cancer tumor cells and play an important role in maintaining the malignant biological characteristics of SKOV-3.