Abstract
The peritoneal fluid (PF) of women with endometriosis contains protease(s) activity able to hydrolyze insulin-like growth factor-binding protein 3 (IGFBP-3), increasing the bioavailability of insulin-like growth factor-1 (IGF-1) locally. Therefore, we characterized the effects of IGF-1 on KLE endometrial-like cells in vitro. The mitogenic effect of IGF-1 was assessed by the analysis of the DNA content and cell count. Apoptosis was triggered experimentally by the 48 hr exposure of KLE cells to 100 nM of adriamycin in the presence and absence of IGF-1 (50 ng/ml). Adriamycin apoptosis of KLE cells was determined by the number of dead KLE cells using trypan blue exclusion and by the DNA fragmentation on simple agarose gel and flow cytometry of propidium iodide and HOECHST 33342-stained KLE cells using an EPICS 753 pulse cytometer. IGF-1 stimulated the growth of KLE cells in a dose-dependent manner (optimal dose of 50 ng/ml) and protected KLE cells from adriamycin (100 nM)-induced apoptosis. These data suggest that IGF-1 is a survival factor for KLE cells. Conceivably, increased IGF-1 activity in the PF can optimize both the survival and ectopic growth of endometrial cells in the peritoneal cavity.
Highlights
Endometriosis is defined as the ectopic growth of endometrial-like tissue on the peritoneum and the organs of the pelvic cavity [1]
KLE endometrial-like cells were characterized in our laboratory previously when we studied their response to growth factors and peritoneal fluid mitogens in parallel experiments using the primary cultures of epithelial cells enzymatically prepared from biopsies of endometrial tissue and endometriotic lesions as control [4,5]
The optimal dose of insulin-like growth factor1 (IGF-1) was 50 ng/ml, which was capable of increasing by 35% the expected DNA content and number of KLE cells (p Ͻ 0.001)
Summary
Endometriosis is defined as the ectopic growth of endometrial-like tissue on the peritoneum and the organs of the pelvic cavity [1]. Recent studies documented that peritoneal fluid (PF) of women with and without laparoscopic evidence of endometriosis contained growth factor activity for endometrial-like cells, which is, at least in part, associated with the production of N-terminal proteolytic fragments of IGF-binding protein 3 (IGFBP-3) [4,5]. Anti-IGF-1 therapy reintroduced an objective clinical response to androgen ablation therapy of prostate cancer patients who had progressed to the hormone refractory stage of the disease [17] These data indicated that exogenous IGF-1 and/or endogenously increased IGF-1 bioavailability in host tissues can exert a novel survival factor activity, rescuing metastatic cancer cells from apoptosis in vitro [18]. Increased IGF-1 activity in the PF can optimize both the survival and ectopic growth of endometrial cells in the peritoneal cavity
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