Anti-cancer effects of alpha lipoic acid, cisplatin and paclitaxel combination in the OVCAR-3 ovarian adenocarcinoma cell line.

BACKGROUND: Ovarian cancer is the leading cause of gynecological cancer deaths. One of the major challenges in treating ovarian cancer with chemotherapy is managing the resistance developed by cancer cells to drugs, while also minimizing the side effects caused by these agents In the present study, we aimed to examine the effects of a combination of alpha lipoic acid (ALA), with cisplatin and paclitaxel in ovarian cancer(OVCAR-3). METHODS: The cytotoxic effects of ALA, cisplatin and paclitaxel on OVCAR-3 cells were determined. Four groups were formed: Control, ALA, Cisplatin + Paclitaxel, ALA + Cisplatin + Paclitaxel. The effects of single and combined therapy on cell migration, invasion and colony formation were analyzed. Changes in the expression of genes related to apoptosis, cell adhesion and cell cycle were analyzed with Real-time polymerase chain reaction(RT-PCR). The oxidative stress index and The Annexin V test were performed. RESULTS: The reduction in rapamycin-insensitive companion of mTOR(RICTOR) expression in the ALA + Cisplatin + Paclitaxel group was found statistically significant(p < 0.05). The decrease in MMP-9 and - 11 expressions the ALA + Cisplatin + Paclitaxel group was statistically significant(p < 0.05). The lowest values for mitogen-activated protein kinase(MAPK) proteins were found in the ALA + Cisplatin + Paclitaxel group. No colony formation was observed in the Cisplatin + Paclitaxel and ALA + Cisplatin + Paclitaxel groups. The lowest wound healing at 24 h was seen in the ALA + Cisplatin + Paclitaxel group. CONCLUSIONS: This study is the first one to investigate the combined treatment of ALA, Cisplatin, Paclitaxel on OVCAR-3. While ALA alone was not effective, combined therapy with ALA, has been found to reduce cell invasion, especially wound healing in the first 24 h, along with tumor cell adhesion.

Relationship between embryo development and apoptotic gene expression of cumulus cells in poor responders and polycystic ovary syndrome.

RESEARCH QUESTION: Is there a relationship between embryo quality, pregnancy rates and apoptotic gene expression in cumulus cells of oocytes collected from patients with poor ovarian response and polycystic ovary syndrome? DESIGN: Fifty infertile couples who underwent assisted reproductive technology treatment were included in the study (Approval date 4 February 2020, number 03). The patients were divided into four group: control (n = 9; 90 oocytes), unexplained infertility (n = 8; 86 oocytes), polycystic ovary syndrome (PCOS) (n = 6; 137 oocytes) and poor ovarian response (POR) (n = 27; 124 oocytes). Cumulus cells were isolated individually from 437 oocytes obtained. Intracytoplasmic sperm injection was undertaken on 365 mature oocytes. The embryos were monitored. Caspase-3, Bax and Bcl-2 gene expressions of the cumulus cells were measured by real-time polymerase chain reaction. RESULTS: A significant and negative correlation was found between Bax and Bcl-2 expressions of the cumulus cells of poor-quality embryos. The increase in Caspase-3 gene expression in the POR group statistically decreases the pregnancy rates. Fertilization and good-quality embryo development of 365 oocytes whose cumulus cells were examined, however, were not associated with apoptotic gene expression. The Bax/Bcl-2 ratio was found to be significantly lower in cumulus cells of mature oocytes. CONCLUSIONS: Our results demonstrated no significant associations between fertilization, quality embryo development and apoptotic gene expression. Bax expression and the Bax/Bcl-2 ratio are high in immature oocyte cumulus cells has shown us that the apoptotic process may begin when the cumulus-oocyte connection exists.

Effects of different doses of melatonin on rat adipose derived mesenchymal stem cells.

OBJECTIVES: Stem cell treatment is based on Melatonin which is crucial for lots of pathological and physiological pathways. Our aim is determining the most appropriate dose of melatonin affecting the rat adipose tissue mesenchymal stem cells. METHODS: Stem cells were isolated from male rat adipose tissue. Differentiation and characterization experiments were performed. Cell viability analyses in stem cells were used the XTT [2,3-Bis-(2-methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide] assay. After 24 h incubation, different concentrations (0.5, 1, 5, 10, 50 µM) of extract were treated to the stem cells for 24 h, 48 and 72 h considering time and dose dependent manner. Total antioxidant status (TAS) and the total oxidant status (TOS) in control cells and melatonin treated cells (5, 10 µM) were determined Rel Assay commercial kits. RESULTS: In 24 h, melatonin increased cell viability in all groups. When we evaluate the effect of melatonin in 48 h, the most proliferation increase was seen at 5, 10 µM doses. When the total oxidant activity melatonin was found to be significantly lower in 5 and 10 µM dose groups of melatonin. CONCLUSIONS: Melatonin increases the survivor of stem cells and the most effective dose is 5 and 10 µM. The reduction of the oxidative stress index as a result of treating melatonin to mesenchymal stem cells showed that melatonin is a powerful antioxidant for stem cells.

Boric acid as a promising agent in the treatment of ovarian cancer: Molecular mechanisms.

PURPOSE: The aim of this study is to determine the therapeutic effects of boric acid cell proliferation, invasion, migration, colony formation, cell cycle and apoptosis mechanisms in ovarian cancer cell line under in vitro conditions. METHODS: MDAH-2774 ovarian cancer cells were employed. Real-time PCR test was used to investigate changes in genes and proteins of cell cycle and apoptosis and identified miRNAs under the addition of boric acid. The apoptosis rates were calculated by TUNEL assay. Matrigel invasion, colony formation and Wound healing tests were used to determine invasion and migration. Oxidative stress index value was calculated for oxidative stress. RESULTS: Boric acid inhibited cell proliferation, invasion, migration and colony formation, but induces apoptosis and oxidative stress. Also, the expression of miRNA-21, miRNA-200a, miRNA-130a and mi-RNA-224 (which are indicators of poor prognosis of ovarian cancer) decreased significantly. CONCLUSION: The potential of boric acid as a natural molecule may supports its effectiveness in reducing adverse effects arising from conventional ovarian cancer treatments.

The effect of adipose-derived mesenchymal stem cell treatment on mTOR and p-mTOR expression in ovarian damage due to cyclophosphomide.

Our aim is to investigate the effect of the Mesenchymal stem cell (MSC) administration on the release of Mammalian Target of Rapamycin (mTOR) and Phosphorylated- mTOR(p-mTOR) in Cyclophosphomide (CTX) induced ovarian damage. Rats divided into three groups. The first group was categorized as the control(C group;n = 6), the second group as CTX-administered group (CTX group;n = 6), and the third group as CTX and MSC-administered group (CTX + SC group;n = 6). CTX was injected intraperitoneally at 50 mg/kg on the first day and at 8 mg/kg during the following 13 days. In Group 3, adipose-derived MSCs (5 × 104) were injected locally into the ovary. Both ovaries were removed at the end of the 8th week. The follicle count was made. The expression of mTOR and p-mTOR was analyzed immunohistochemically. The follicles in the ovary of Group C were observed in normal structures. Degeneration was evident in the CTX group. In the CTX + SC group, the degenerative appearance monitored in the CTX group vanished in most areas, and fibrosis was greatly reduced. The number of follicles in the CTX group was lower than that of both C and CTX + SC groups (p < 005). In the C group, mTOR showed strong positive staining while mTOR and p-mTOR expression was negative in all follicles in the CTX group. Both mTOR and p-mTOR revealed moderate positive expression in the CTX + SC group. MSC therapy rescued the damage ovarian function created by CTX, reducing follicle loss. MSCs were shown to inhibit the loss of mTOR and p-mTOR signaling, which is key to meiosis in oocytes.

Autogenous Tooth Bone Graft and Simvastatin Combination Effect on Bone Healing.

OBJECTIVE: Autogenous tooth bone grafts (ATGM) are materials prepared from extracted teeth and have been used for bone augmentation. These graft materials are known to have similar structures and components to bone grafts. In this sense, this study aimed to evaluate all the tooth layers mixed with simvastatin without any demineralization process effect on bone formation. METHODS: In 60 Wistar albino rats, a standardized 6.0 m-diameter critical size bone defect was created in their calvarium. The study consists of 1 control and 4 experimental groups. In the control group (12 rats), the defects were left empty. The defects were grafted only with ATGM in Group 1, with ATGM mixed with simvastatin in Group 2, autogenous bone graft mixed with simvastatin in Group 3, and with xenogenic bone graft mixed with simvastatin in Group 4. The animals were sacrificed at the 7th and 28th days after operation. RESULTS: PCR, micro CT and histological results show that bone formation was enhanced in the experimental groups in comparison to the control group. Group 1 and Group 2 had similar bone formation rate when compared to Group 3 and Group 4 at the 28th day after operation. CONCLUSION: This study concludes that mineralized teeth may be used for defect reconstruction without any demineralization process. Autogenous mineralized tooth bone graft should be mixed with simvastatin for bone regeneration like other grafts.

Adipose derived mesenchymal stem cell treatment in experimental asherman syndrome induced rats.

Asherman syndrome (AS) occurs due to fibrosis or uterine adhesions as a result of damage to the basal layer of the endometrium. The aim of this study is investigating the effects of adipose tissue-derived mesenchymal stem cell (ADMSC) application on the expression of vascular endothelial growth factor (VEGF), insulin-like growth factor (IGF-1), miRNA-98, miRNA199a in endometrial tissue in rats with AS. Study groups were designed as, control (C), Asherman syndrome (AS), AS + oral estrogen (ASO), AS + ADMSC (ASSC), AS + oral estrogen + ADMSC (ASSCO) with 7 samples in each group. Characterization and differentiation experiments were performed in ADMSC obtained. Two weeks after the development of the AS, ADMSC therapy was applied. BrdU (5-bromo-2'-deoxyuridine) labeling was performed to show the presence of ADMSC in the tissues. Rats were sacrificed after 8 weeks and bilateral uterine horn resection was performed. Tissues were fixed in formaldehyde. After routine tissue follow-up, sections were taken and evaluated with hematoxylin eosin staining. VEGF1 and IGF1 expressions were evaluated by immunohistochemical staining and western blot analysis. Expression changes of miR-98 and miR-199a were detected by RT-PCR. Our results showed that stem cells and estrogen giving together reduced inflammation and fibrosis in the endometrium. Immunohistochemistry and western blot results suggested that this effect was achieved especially through IGF-1. In our study, decreased miR-98 and miR-199a expressions were determined in Asherman syndrome. Furthermore, no changes of miRNA expressions were observed in treatment groups.