Significance of tumor-associated protein detection by flow cytometry in ovarian cryopreservation and transplantation in nude mice bearing human epithelial ovarian cancer

Abstract

Objective To investigate screening criteria of paraneoplastic normal ovarian tissue and the feasibility of human ovarian cryopreservation and transplantation in nude mice by detecting tumorassociated protein.Methods Human epithelial ovarian cancer cell line OVCAR3 were grown in subcutaneous tissue and the tumor tissues were orthotopic implanted.The expressions of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 were detected by flow cytometry in tumor tissues,tissues of proximal,median and distal adjacent to tumor,as well as normal ovarian tissues of nude mice.The tissues of group with all negative iudicators,group with negative expressions of CK-7 CA125 Survivin,group with positive expressions of CK-7 CA125 Survivin,cancer tissues and normal ovarian tissues of nude mice were used for ovarian cryopreservation and transplantation.The carcinogenesis rate and CA125 level were analyzed in each group after transplantation.The CA125 level was also detected under different severities after orthotopic implantation in nude mice.Results 46 samples ovarian tissues with normal biopsy were gained from 52 cases of human epithelial ovarian cancer of orthotopic implantation model in nude mice.The CA125 level of only ovarian inoculation group was higher than those of control group (P ＜ 0.01),and lower than those of outside ovarian inoculation group (P ＜ 0.05).The expression rates of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 in tumor tissues were 93.3 ％ (28/30),93.3 ％ (28/30),86.7 ％ (26/30),86.7 ％ (26/30),83.3 ％ (25/30),83.3 ％ (25/30) respectively.They were 70.0 ％ (21/30),70.0 ％ (21/30),63.3 ％ (19/30),63.3 ％ (19/30),60.0 ％ (18/30),56.7 ％ (17/30) in proximal tissues adjacent to tumor respectively,and they were 33.3 ％ (10/30),30.0 ％ (9/30),30.0 ％ (9/30),30.0 ％ (9/30),26.7 ％ (8/30),23.3 ％ (7/30) respectively in median tissues adjacent to tumor.In distal tissues adjacent to tumor were 26.7 ％ (8/30),23.3 ％ (7/30),26.7 ％ (8/30),26.7 ％ (8/30),30.0 ％ (9/30),30.0 ％ (9/30) respectively.The expressions of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 were all negative in 20 samples normal ovarian tissues adjacent to tumor.The expressions of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 in proximal tissues adjacent to tumor were lower than those in tumor tissues (P ＜ 0.05),and higher than those in median and distal tissues adjacent to tumor (P ＜ 0.01).The expressions were no difference between in median and distal tissues (P ＞ 0.05).The strong positive expressions of CK-7,CA125,Survivin were higher than those of p53,MMP-2 and TIMP-2 (P ＜ 0.05).The expressions of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 were all negative in 20 samples normal ovarian tissues of nude mice.If all indicators or the main indicators,like CK-7,CA125 and Surrv ivin were not expressed,the tissues did not develop canceration after transplantation.And the carcinogenesis rate and serum CA125 level were lower than that of the group with positive expressions of CK-7,CA125 and Survivin (P ＜ 0.05).Conclusions The expressions of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 show decreasing trend to non-cancer direction.Negative expressions of all these tumor-associated genes or the main indicators,CK-7,CA125 and Survivin can be organized as the screening criteria of security of ovarian tissues for transplantation.Flow cytometry can be used as the effective detection method.Ovarian cryopreservation and transplantation in nude mice bearing human epithelial ovarian cancer is feasible and safe. Key words: Ovarian neoplasms; Cryopreservation; Transplantation; Genes, neoplasm; Paraneoplasm tissue