Flavopiridol Induces Apoptosis via Mitochondrial Pathway in B16F10 Murine Melanoma Cells and a Subcutaneous Melanoma Tumor Model

Flavopiridol is a cyclin-dependent kinase (CDK) inhibitor that promotes cell cycle arrest. We aimed to examine the anti-proliferative effects of the flavopiridol and oxaliplatin combination on p16INK4A deficient melanoma cells B16F10 and also its apoptotic effects on a subcutaneously injected B16F10 allograft melanoma tumor model. Flavopiridol and oxaliplatin treated B16F10 cell viability was determined by MTT assay. C57BL6 mice were injected with B16F10 cells and treated with flavopiridol after tumor implantation. BRAF and BCL2L1 mRNA expression levels were measured using reverse transcription-polymerase chain reaction (RT-PCR). Caspase 9 and caspase 3/7 activity were determined by activity assay kits. Proliferating cell nuclear antigen (PCNA) and B-cell lymphoma 2 (BCL-2) protein expression levels were analyzed immunohistochemically. Flavopiridol and oxaliplatin decreased cell death. Flavopiridol enhanced caspase 3/7 and caspase 9 activities in vitro and in vivo in a dose dependent manner via the mitochondrial apoptotic pathway. Even though there was a significant increase in Bcl-2 staining, PCNA staining was decreased in flavopiridol-administered mice. Decreased PCNA expression showed antiproliferative effects of flavopiridol which might be the result of cell-cycle arrest. Flavopiridol can be used as a cell cycle inhibitor, which induced apoptosis through the mitochondrial pathway, independently from BCL2 in B16F10 cells and B16F10 injected C57BL6 allografts. </p

Screening of postpartum depression among new mothers in Istanbul: a psychometric evaluation of the Turkish Edinburgh Postnatal Depression Scale

OBJECTIVE:This study is the first concerted effort to ascertain factor structure of EPDS using evidence based analytical techniques. It is the most widely used scale for assessing postpartum depression in Turkey, and yet no investigations have been conducted to assess it factor structure. This study was conducted from April 2012 to April 2018 at the Marmara University Hospital operating under the name of Marmara University Pendik Training and Research Hospital in Istanbul Turkey. RESULTS:A total of 1700 women were included in this study, who responded to the EPDS, in addition to demographic characteristics and well-being of their offspring. A total of 1615 mothers provided adequate data for inclusion in analysis. Standardized Chronbach's alpha for EPDS was 0.81 with corrected item-total correlations ranging from 0.35 to 0.62. Parallel analysis, MAP Velicer Test and Hull's method dictated retaining of one factor structure. All the items revealed adequate communalities (> 0.20) except item 2 (enjoyment) and item 10 (self-harm). Their communalities were 0.16 and 0.19, however, these items were not dropped. All of the items yielded moderate to strong factor loadings. Minimum factor loading was for item 2 (0.40) and highest for item 8 (0.71)

Alcohol use disorder and emotional abuse: the mediating role of early maladaptive schemas

WOS: 000466903900002Objective: Studies have shown a relationship between the development of alcohol use disorder (AUD) and the existence of childhood traumas (CT). In this study, in order to gain a better understanding of the association between CT and AUD, the relationship between CT and AUD was analyzed. Further, it was investigated whether Early Maladaptive Schemas (EMS) play a mediating role in the relationship between emotional abuse (EA) and AUD. Methods: The participants were consecutively admitted male alcohol-dependent inpatients (n=220) and healthy controls (n=108). The participants were investigated with the Childhood Trauma Questionnaire (CTQ-28), Young Schema Questionnaire-Short Form (YSQ-SF) and Michigan Alcoholism Screening Test (MAST). Mediation analyses were conducted in order to evaluate the mediating effect of EMS between the relationship of AUD and CT. Results: The difference between CT total score and the EA score between the control group and the AUD group were statistically significant. An analysis of different types of traumas in the correlation with AUD revealed EA as the main predictor of AUD both directly and through its mediating effect with the EMS. In addition, it was determined that in the relationship between EA and AUD, EMS such as Enmeshment, Emotional Inhibition, Dependence and Insufficient Self-Control had mediating effect. Discussion: CT, especially EA are quite common in male inpatients with AUD and the negative effects of EA may play an important role in the development of AUD by causing EMS. Our findings imply that those with EA history in childhood may be particularly vulnerable to the development of AUD in adulthood. Therefore, in the prevention of the development and in the treatment of AUD, it may be useful to take into consideration EA experiences, to screen EMS that may mediate such relationship through scales and to conduct interventions in this regard

Flavopiridol's antiproliferative effects in glioblastoma multiforme.

Glioblastoma multiforme (GBM) is largely refractory to surgical operation, radiotherapy, and chemotherapy in use today. Remaining lifetime accounting for the GBM-affected patients varies between 12 and 16 months generally. The most frequently altered genes in GBM are p53, epidermal growth factor receptor, PTEN, and cyclin-dependent kinase inhibitor 2A. Our aim is to investigate the antiproliferative and apoptotic effects of flavopiridol, a cyclin-dependent kinases and specific phosphokinase inhibitor, on glioblastoma cell lines having different genetic profiles: U87MG, U118MG, and T98G

Effects of thiostrepton alone or in combination with selumetinib on triple-negative breast cancer metastasis

Objective FoxM1 transcription factor contributes to tumor metastasis and poor prognosis in many cancers including triple-negative breast cancer (TNBC). In this study, we examined the effects of FoxM1 inhibitor Thiostrepton (THIO) alone or in combination with MEK inhibitor Selumetinib (SEL) on metastatic parameters in vitro and in vivo. Methods Cell viability was determined by MTT assay. Immunoblotting and immunohistochemistry was used to assess metastasis-related protein expressions in 4T1 cells and its allograft tumor model in BALB/c mice. In vivo uPA activity was determined by enzymatic methods. Results Both inhibitors were effective on the expressions of FoxM1, ERK, p-ERK, Twist, E-cadherin, and Vimentin alone or in combination in vitro. THIO significantly decreased 4T1 cell migration and changed the cell morphology from mesenchymal-like to epithelial-like structure. THIO was more effective than in combination with SEL in terms of metastatic protein expressions in vivo. THIO alone significantly inhibited mean tumor growth, decreased lung metastasis rate and tumor foci, however, no significant changes in these parameters were observed in the combined group. Immunohistochemically, FoxM1 expression intensity was decreased with THIO and its combination with SEL in the tumors. Conclusions This study suggests that inhibiting FoxM1 as a single target is more effective than combined treatment with MEK in theTNBC allograft model. The therapeutic efficacy of THIO should be investigated with further studies on appropriate drug delivery systems.Scientific and Technological Research Council of Turkey, (TUBITAK) [1002116S162]; Teaching Staff Training Program of TurkeyThis work was supported by the Scientific and Technological Research Council of Turkey, (TUBITAK) under Grant (number 1002116S162) and Teaching Staff Training Program of Turkey

Relationship between hyperandrogenism, obesity, inflammation and polycystic ovary syndrome

This prospective study aimed to determine the status of circulating levels of C-reactive protein (CRP), tumor necrosis factor alpha (TNF-alpha), IL-27, IL-35, IL-37, alpha-1 acid glycoprotein in patients with polycystic ovary syndrome (PCOS) compared with controls and to evaluate their relation with hyperandrogenism and obesity. Forty-eight patients with PCOS (29 obese, 19 lean) and 40 healthy controls (20 obese, 20 lean) were enrolled. CRP, TNF-alpha, IL-27, IL-35, IL-37, alpha-1 acid glycoprotein, sex hormone-binding globulin (SHBG), dehydroepiandrosterone sulfate (DHEA-S) levels were measured. Levels of total testosterone, A4, DHEA-S were significantly higher in patients with PCOS than in controls both in the obese and lean groups, while levels of SHBG were significantly lower in all patients with PCOS than in all (p < 0.05). Free androgen index (FAI) values were significantly higher in all patients with PCOS than in all controls (all p < 0.05). Levels of CRP, TNF-alpha, alpha-1 acid glycoprotein were significantly increased in all patients with PCOS compared with all controls (all p < 0.001). FAI had a positive correlation with CRP, TNF-alpha, alpha-1 acid glycoprotein, a negative correlation with IL-27, IL-25, IL-37 (all p50.01). Body mass index had a negative correlation with IL-27, IL-35, IL-37, a positive correlation with alpha-1 acid glycoprotein, FAI (p < 0.05). The findings confirm the proinflammatory state of PCOS. Moreover, obesity along with PCOS significantly elevates the inflammatory status and hyperandrogenism

Flavopiridol's antiproliferative effects in glioblastoma multiforme

Aim of Study: Glioblastoma multiforme (GBM) is largely refractory to surgical operation, radiotherapy, and chemotherapy in use today. Remaining lifetime accounting for the GBM-affected patients varies between 12 and 16 months generally. The most frequently altered genes in GBM are p53, epidermal growth factor receptor, PTEN, and cyclin-dependent kinase inhibitor 2A. Our aim is to investigate the antiproliferative and apoptotic effects of flavopiridol, a cyclin-dependent kinases and specific phosphokinase inhibitor, on glioblastoma cell lines having different genetic profiles: U87MG, U118MG, and T98G. Materials and Methods: Cell viability and IC50 values were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, protein expressions were determined by Western blot and caspase activities were analyzed by activity kit. Results: Western blot analysis showed down-regulation of the cyclin D1, c-Myc, and p53 protein activities, and up-regulation of p27(KIP1) activity after flavopiridol treatment. Additionally, flavopiridol diminished p-Akt protein levels generally which induces inhibition of proliferation. Conclusion: The present study demonstrated that flavopiridol did not induce caspase-3/7 activation, BIM, and BAX pro-apoptotic proteins but it leads to the expression changes of various proteins that inhibit proliferation and eternity in glioblastoma cell lines which have different genetic alterations