Abstract
Objective Autophagy has been reported to be involved in the development of various disorders such as neurodegenerative and metabolic diseases and tumors. Autophagy activators and inhibitors are also potential therapeutics for these diseases. However, the mechanism of autophagic involvement in different diseases is not the same, and the role of autophagy in endometriosis (EM) has not yet been elucidated. This research investigated the mechanism by which autophagy acts in EM, with the aim of establishing a theoretical basis for its prevention and treatment through the targeted interference with autophagy. Methods We used an RNA interference fragment targeting ATG5, the autophagy activator rapamycin, and the autophagy inhibitor 3-MA or overexpression of filopodia-related protein fascin-1, in conjunction with clonogenic assays, growth curves, and scratch assay to investigate the influence of autophagy on cellular growth, proliferation, and invasiveness. We collected specimens from 20 clinical cases of EM and investigated the protein expression of the autophagic marker LC3-II, the autophagic substrate p62, and fascin-1. Results Rapamycin was able to inhibit the proliferation and colony formation of the endometriotic cell line CRL-7566, whereas the autophagy inhibitor 3-MA as well as the interference with the autophagy-related gene ATG5 had the opposite effect. More importantly, the autophagy activator rapamycin was able to inhibit the growth of filopodia in the endometriotic cells, and the overexpression of the fascin-1 restored the rapamycin-induced decrease of invasiveness. We found that the expression of the autophagy marker LC3-II was significantly reduced among the clinical EM specimens compared to the control group, while the expressions of fascin-1 and autophagic substrate p62 were increased. Conclusion Our results indicate that the inhibition of autophagy and exogenous expression of fascin-1 may promote the invasiveness of endometrial cells. As a corollary, autophagy represents a potential target for the treatment of EM.
Highlights
Endometriosis (EM) denotes the occurrence, growth, and infiltration of glandular and mesenchymal endometrial tissue outside the endometrium and is often characterized by repeated bleeding or nodule formation [1]
We used two different methods to interfere with the autophagy level of endometrial CRL-7566 cells-treatment with the autophagy inhibitor 3-MA and transfection with an RNA interference fragment targeting the ATG5 gene in order to repress autophagy
The results of immunoblotting have shown that the expression level of ATG5 was obviously reduced upon RNA interference (Figure 1(b))
Summary
Endometriosis (EM) denotes the occurrence, growth, and infiltration of glandular and mesenchymal endometrial tissue outside the endometrium and is often characterized by repeated bleeding or nodule formation [1]. It is a frequently encountered gynecological endocrine disease and has a high incidence [1]. Autophagy is one of the major ways in which eukaryotic cells remove abnormal proteins and damaged organelles through lysosomal degradation [6]. It plays a crucial role in maintaining cellular homeostasis
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