Investigation of T-cell receptor rearrangement in tumor tissues in ovarian cancers by means of multiplex PCR method

Object: In this study, clonal selection in patients will be investigated by searching the T-cell receptor rearrangement in over carcinomas. Through the future immunologic studies, the treatment to be applied will be directed depending on the clonal status of patients and a treatment type specific to each patient will be developed. Method: 24 patients with malign adnexal mass and 23 patients with benign adnexal mass who have applied to Ege University Medical Faculty Department of Obstetrics and Gynecology between the dates of November 2010 and November 2012 were included in the study. Fresh tissue samples were taken from primary ovary tumor tissue of the patients diagnosed with over carcinoma (n=24) during the operation. However, beta, gamma and delta genomic DNA was isolated from fresh tissue samples taken from non-neoplastic ovary which have been operated due to the causes related with uterus and underwent prophylactic oophorectomy (n=23) by means of appropriate commercial kits for the cell type. The amount of the isolated genomic DNA samples was measured at a wavelength of 260 nm. The selected high quality DNA samples were determined by PCR method by using Biomed-2 primaries in order to specify T-cell gene clonality. After PRC, heterodublex analysis was made on all samples in order to prevent false positivity and loaded to Polyacrylamide gel electrophoresis system. After the samples were subjected to Polyacrylamide gel electrophoresis with previously optimized voltage and periods, gel dying with EtBr and distilled water wash operations were carried out. Then, the gel was displayed in the gel imaging system and its photo was taken. Size of bands in the photo and analysis ranges were examined and T-cell clonality was evaluated for each patient. Relationship between T cell rearrangement and prognostic variables such as age of patient, stage, histopathological type of tumor, tumor grade, tumor diameter etc. was evaluated. Findings: TCRB-A positivity ratio showed no significant difference (p˃0,05) in the study and control groups. TCRB-B positivity ratio was found to be significantly higher in the study group compared to the control group (p<0,05). However, TCRB-C showed no significant difference in control and study groups (p˃0,05). In the control and study groups, TCRG-A positivity ratio showed no significant difference (p ˃ 0,05). In the control and study groups, TCRG-B positivity ratio showed no significant difference (p ˃ 0,05). TCR-D positivity ratio was found to be significantly higher in the study group compared to the control group (p<0,05). Result: Sub-groups of TCR were shown in the ovary cancer case series. The significant difference in TCRB-B and TCR-D sub-groups will provide an insight to immunotherapy.