Objective. To determine the effects of Mullerian inhibiting substance (MIS) treatment on endometriosis cells through study of apoptosis and autophagy. Design. Experimental in vitro study. Setting. University research laboratory. Cell Line. CRL-7566 endometriosis cell line. This line was established from a benign ovarian cyst taken from a patient with endometriosis. Interventions. In vitro treatment with MIS. Main Outcome Measures. The main outcome measures were cellular viability, proliferation, cell-cycle arrest, and induction of apoptosis and autophagy in endometriotic cells. Results. MIS treatment inhibited proliferation of endometriosis cells and induced apoptosis, as indicated by Annexin V staining, and induced caspase-9 cleavage and cell-cycle arrest, as evidenced by increased expression of p27 CDK-inhibitor. MIS treatment also induced autophagy in endometriosis cells as demonstrated by a significant increase in LC3-II induction, a hallmark of autophagy. Conclusions. MIS inhibits cell growth and induces autophagy, as well as apoptosis, in ectopic endometrial cell lines. Our results suggest that MIS may have a potential as a novel approach for medical treatment of endometriosis. Further studies may be needed to test the efficacy of MIS treatment in animal models and to develop MIS treatment specifically targeted to the endometriosis. 1. Introduction Endometriosis, one of the most common and debilitating diseases in women, is characterized by endometrial-like tissue outside the uterus. As a major cause of pelvic pain and infertility, it affects 6% to 10% of women of reproductive age, 50% to 60% of women and teenage girls with pelvic pain, and up to 50% of women with infertility [1]. Endometriosis also has a great impact on healthcare system. Annual healthcare costs and costs of productivity loss associated with endometriosis were estimated at $22 billion in 2002 [2]. Current treatments for endometriosis have their shortcomings. Long-term treatment of patients with endometriosis involves repeated courses of medical therapy, surgical therapy, or both. Medical modalities include nonsteroidal anti-inflammatory drugs (NSAIDs), combined contraceptive pills, oral progestogens, depot medroxyprogesterone acetate, danazol, and GnRH agonists. In addition, aromatase inhibitors have been reported to be efficacious in some observational trials, but data are insufficient to recommend their routine use. Mainstays of surgical treatment include laparoscopic ablation of endometriotic lesions, surgical excision of large endometriomas, and unilateral or bilateral
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