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An Experimental Study on Expression of HLA-G in Ovarian Cancer Cell Line SKOV-3

Author: WangLin
Tutor: DongJianChun
School: Shandong University
Course: Obstetrics and Gynaecology
Keywords: Human leucocyte antigen-G SKOV-3 cell line JEG-3 cell line ovarian neoplasms
CLC: R737.31
Type: Master's thesis
Year: 2008
Downloads: 34
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Abstract


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.

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CLC: > Medicine, health > Oncology > Genitourinary tumors > Female genital tumors > Ovarian tumors
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