Effect of GnRH analogues and octreotide treatment on apoptosis and the cell proliferation of endometrium adenocarcinoma cell lines.

OBJECTIVE The aim of this study was to compare apoptotic and antiproliferative effects of gonadotropin-releasing hormone analogues and their combination with octeotide on endometrioid endometrial cancer cell lines. MATERIAL AND METHOD Women diagnosed with endometrioid adenocarcinoma at the department of Gynecology and Obstetric of Kocaeli University Medical School were included in this research. Endometrium cancer cell lines obtained from three patients were used for this study. After trypsinization in 0.5% in calcium magnesium, free phosphate buffer solution (CMFPBS) cells were seeded on glass slides in 24-well plates containing DMEM-F12 medium and 10% fetal calf serum as culture medium. Cells were incubated for 24 hours at 37ºC in 5% CO2. GnRH agonist leuprolide (Lucrin 1 μmol/L), GnRH antagonist ganirelix (Orgalutran 1 μmol/L), leuprolide with octreotide (Sandostatin 10-6 mol/L), ganirelix with octreotide and no drug were added to the wells. Apoptosis and cells proliferations were evaluated after 12, 24, 48 and 72(th) hours of incubation. The percentage of apoptotic cells was evaluated by TdT mediated biotin-dUTP nick-end labeling (TUNEL) method; cell proliferation was assessed by bromodeoxyuridine (BrdU) incorporation. RESULTS Apoptotic index in grade I EEC cell line among ganirelixoctreotide treated cells and leuprolide-octreotide combination therapy were respectively higher than the untreated control (p<0.001, p=0.001). The number of apoptotic cells in grade II EEC cell line among leuprolide-octreotide and leuprolide were significantly (p<0.001, p<0.001) higher than in controls. In grade III EEC cell line, the number of TUNEL positive cells among leuprolide, ganirelix and ganirelixoctreotide therapiy groups were significantly higher than in untreated control. Time dependent antiproliferative effect was obtained with leuprolide and leuprolide-octreotide in grade I EEC (p<0.001, p<0.001). Grade II EEC cell line is not influenced by hormonotherapies. However, the antiproliferative effect was obtained with ganirelix, leuprolide and leuprolide-octreotide in grade III cell line. CONCLUSION GnRH analogues appears to have a direct effect, enhancing the apoptotic index and decreasing the cell proliferation in endometrial adenocancer cell lines.