Understanding Benign Prostatic Hyperplasia (BPH)
E-mail address: sachikoma aol. Use the link below to share a full-text version of this article with your friends and colleagues. Learn more. A high recurrence rate after medical treatment is a major clinical problem for patients with endometriosis.
Combined treatment with MK and chloroquine markedly reduced cell growth and regrowth after discontinuation of treatment in endometriotic stromal cells compared BPH Chirurgie Recovery-Tour cells treated with either drug alone.
Furthermore, combined treatment reduced the size of endometriotic implants, whereas no effects on endometriotic implants treated with either drug alone were observed in a mouse xenograft model of endometriosis. The present findings suggest that a novel strategy for treatment of endometriosis may involve decreasing the number of endometriotic cells that can survive treatment and then preventing regrowth by autophagy inhibition.
A high recurrence rate after medical treatment, with or without surgery, is a major clinical problem for BPH Chirurgie Recovery-Tour with endometriosis Guo, Establishment of a complete cure for patients with endometriosis depends on new targets and strategies. Until now, numerous studies have evaluated whether candidate molecules can decrease the number of cells that survive treatment in in vitro cell culture systems Soares et al. However, it is BPH Chirurgie Recovery-Tour critical to evaluate whether candidate molecules for the treatment of endometriosis can prevent relapse of the disease after treatment is discontinued Matsuzaki et al.
However, a high proliferation rate of DES cells was observed after drug discontinuation Matsuzaki et al. Our previous findings suggested that MK may induce autophagy, which may inhibit cell death. In our previous study, we further evaluated the effects of chloroquinean antimalarial drug and an autophagy inhibitor Matsuzaki et al.
Both chloroquine and hydroxychloroquinethe hydroxylated analogue of chloroquineare the only autophagy inhibitors approved by the Food and Drug Administration Yang et al. Autophagy plays a dual role in various BPH Chirurgie Recovery-Tour by promoting both cell survival and cell death Codogno and Meijer, ; Das et al.
Our previous study suggested that autophagy induced by MK may play a cytoprotective role in endometriosis Matsuzaki et al. To date, many studies have shown that various types of chemotherapy and radiation therapy induce autophagic cytoprotective functions in tumour cells, and inhibition of autophagy BPH Chirurgie Recovery-Tour chloroquine or hydroxychloroquine is effective for chemosensitization and radiosensitization Janku et al.
Several studies have shown that chloroquine or hydroxychloroquine sensitizes cells to the actions of MK, with or without conventional chemotherapeutic agents in various cancer cell types Cheng et al. In addition, studies have shown that treatment with chloroquine or hydroxychloroquine alone is effective in different cancers through targeting of basal autophagy Kimmelman, ; BPH Chirurgie Recovery-Tour et al.
A BPH Chirurgie Recovery-Tour study showed that treatment with hydroxychloroquine alone decreased lesion numbers and disrupted lesion histopathology in an autologous mouse model of endometriosis Ruiz BPH Chirurgie Recovery-Tour al.
These previous findings along with those of our group Matsuzaki et al. Therefore, we first performed a comparative in vitro study prior to validation in animal experiments.
Currently available pharmacological inhibitors of autophagy are not entirely specific Yang et al. BPH Chirurgie Recovery-Tour studies should be combined with genetic approaches to more specifically inhibit the autophagy pathway Barth et al.
All BPH Chirurgie Recovery-Tour were performed in accordance with the approved guidelines and regulations. Informed written consent was obtained from each patient prior to tissue collection. Endometrial and endometriotic samples from 60 patients who had histological evidence of rectovaginal deep infiltrating endometriosis DIE were used for the present analysis. In addition, endometrial tissues from 15 patients without BPH Chirurgie Recovery-Tour tubal infertility were used BPH Chirurgie Recovery-Tour controls.
All tissues were derived from the BPH Chirurgie Recovery-Tour phase of the menstrual cycle. Cells at passage 1 were used for experiments. The polyacrylamide gel was then polymerized and dried onto the flexible plastic support. Then, one polyacrylamide gel was placed in each well.
The polyacrylamide gel was coated with type I collagen 0. Then, the gels were BPH Chirurgie Recovery-Tour in complete medium overnight to hydrate and equilibrate. Per cent cell proliferation was calculated as per cent of vehicle control.
Both early apoptotic Annexin V positive and PI negative and late Annexin V positive and PI positive apoptotic cells were included in cell death determinations, as previously described Matsuzaki et al. Blots were processed as described in the SNAP i. The Western blot bands were quantified using ImageJ software version 1.
Results were corrected by actin to normalize the loading. The number of LC3 or p62 puncta per cell was quantified using ImageJ software version 1. The average number of LC3 or p62 puncta per cell was quantified in at least cells for each group using ImageJ software version 1. The experiment was conducted under a licence from the French Ministry of Agriculture.
Mice had free access to food and water. After completion of the experiment, all mice were killed with an anaesthetic overdose. BPH Chirurgie Recovery-Tour nude mouse model of endometriosis was used as previously described Matsuzaki and Darcha, b Nude mice were implanted s.
All procedures were performed under isoflurane anaesthesia. On day 14, mice were randomized into four experimental groups.
Endometriosis is characterized by dense fibrotic tissues Giudice and Kao, Intraperitoneal injections were given under isoflurane anaesthesia. Treatment regimens were determined during preliminary experiments Supplementary methods.
For sample analysis, each animal was BPH Chirurgie Recovery-Tour on the tail using nontoxic permanent markers, and researchers were blinded to treatment groups. Volumes of endometriotic implants were measured by direct measurement BPH Chirurgie Recovery-Tour calipers.
Then, volume change after treatment percentage of volume on day 21 relative to that on day 15 within the same mice was calculated for each mouse. Immunohistochemical staining BPH Chirurgie Recovery-Tour performed on paraffin sections with a rabbit monoclonal antibody directed against pAktCell Signaling Technologya rabbit polyclonal antibody directed against LC3MBL or a rabbit polyclonal antibody directed against p62MBL.
The data and statistical analysis comply with the recommendations on experimental design and analysis in pharmacology Curtis et al. For ANOVA, the post hoc test was run only if F achieved statistical significance and no significant variance inhomogeneity was observed. Previous studies including ours showed that it is critical to model in vivo tissue compliance conditions in vitro to fully investigate cell responses to drugs Zustiak et al. Therefore, we evaluated the effects of substrates of varying stiffness on inhibition of cell proliferation.
During the experimental period, all mice survived, and no significant differences in body weight were observed among the four experimental groups. All of BPH Chirurgie Recovery-Tour mice in the present study developed histologically confirmed endometriotic lesions with glandular structures and stroma.
Numerous studies have shown that endometrium of patients with endometriosis may differ biochemically from that of patients without endometriosis Giudice and Kao, In the present study, all of the mice developed histologically confirmed endometriotic lesions with glandular structures and stroma.
Because the duration of the combined treatment in vivo was longer than that in vitroa potential explanation for the discrepancy between the in vitro and in vivo findings may be that a longer duration of combined treatment might significantly induce apoptosis in BPH Chirurgie Recovery-Tour. In contrast to the present BPH Chirurgie Recovery-Tour, hydroxychloroquine treatment alone before implantation of uterine fragments onto peritoneum disrupts endometriotic epithelial cells but does not significantly affect BPH Chirurgie Recovery-Tour size of the endometriotic implants Ruiz et al.
A limitation of the present mouse model is that it is not possible to investigate whether endometriotic implants can regrow BPH Chirurgie Recovery-Tour whether autophagy is required for regrowth after discontinuation of treatment.
Currently, no small animal models of endometriosis are available to investigate recurrence after medical treatment Vanhie et al. Development of such models that closely mimic the clinical scenario of endometriosis could greatly contribute to the development of novel therapeutic strategies in patients with endometriosis.
The present clonogenic assay showed that chloroquine alone BPH Chirurgie Recovery-Tour significantly affected survival of EES cells, compared with that of DES cells from the same patient. In addition, chloroquine alone had no significant effect on the growth inhibition of endometriotic implants in the present mouse model of endometriosis, when treatment was started after development of endometriotic implants.
These findings do not support the use of chloroquine or hydroxychloroquine alone for the treatment of patients who have already had endometriosis. However, hydroxychloroquine alone before implantation of uterine fragments onto peritoneum decreased lesion numbers in BPH Chirurgie Recovery-Tour mouse model of endometriosis Ruiz et al. Further studies are thus required to BPH Chirurgie Recovery-Tour whether chloroquine or hydroxychloroquine alone could BPH Chirurgie Recovery-Tour an effective novel strategy for prevention of the development of endometriosis.
Autophagy can be also blocked upstream of autophagosome formation, resulting in a limited number of autophagosomes and reduced autophagic activity Galluzzi et al. The present results revealed no significant change in ATG12 protein expression after treatment with either chloroquine alone or MK alone. Canonical autophagy may maintain BPH Chirurgie Recovery-Tour EES cell proliferation after discontinuation of drug treatment.
In DES cells, activation of canonical autophagy may be required for cell regrowth, when the number of cells that can survive BPH Chirurgie Recovery-Tour treatment is sufficiently decreased. Studies have shown that the role of autophagy may differ between cancer stem BPH Chirurgie Recovery-Tour CSCs and the bulk cancer cell population Gong et al.
Endometriosis, although a benign disease, shares many aspects BPH Chirurgie Recovery-Tour cancer. A growing body of evidence suggests that endometriosis originates from endometrial stem or progenitor cells Gargett et al.
Autophagy may be required for maintenance of endometriotic stem cells that can BPH Chirurgie Recovery-Tour drug treatment. The present results suggest that a novel treatment strategy for patients with endometriosis could involve decreasing the number of cells that can survive treatment and then preventing regrowth by autophagy inhibition.
However, autophagy is critical for BPH Chirurgie Recovery-Tour physiological and pathological processes Mizushima, ; Mizushima et al. The present study showed that autophagy inhibition enhanced regrowth of EES cells after discontinuation of drug treatment. Inhibition of autophagy may increase the risk of healthy endometrial tissues undergoing malignant transformation.
In addition, autophagy is also critical for optimal immune function Levine and Kroemer, As recently proposed by Galluzzi et al. However, to date, only a few studies have investigated the role of autophagy in endometriosis, as well as in the endometrium, and the results remain controversial Choi et al. We are most grateful to all the patients who participated in the present study.
All authors read and BPH Chirurgie Recovery-Tour the final version of the paper. This Declaration acknowledges that this paper adheres BPH Chirurgie Recovery-Tour the principles for transparent reporting and scientific rigour of preclinical research recommended by funding agencies, publishers and other organisations engaged with supporting research.
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