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Objective Pancreatic cancer is a leading cause of cancer-related deaths in developed countries, with a 5-year average survival rate of less than 5%, making it a malignant disease. Gemcitabine (GEM), an FDA-approved pyrimidine antimetabolite, is widely used in pancreatic cancer treatment. However, due to its targeting of all dividing cells, severe side effects are frequently observed in patients undergoing GEM treatment for pancreatic cancer. Consequently, meta-analyses have shown that the combination of GEM with other active compounds significantly improves the 1-year survival rate of pancreatic cancer patients. Epigallocatechin3-gallate (EGCG), an active compound found in green tea (Camellia sinensis), has proven anticancer activity in pancreatic cancer. Subsequent studies have demonstrated that EGCG enhances the sensitivity of pancreatic cancer cells to GEM. However, among the studies conducted to date, the impact of the combination of EGCG and GEM on the expression of critical microRNAs, which act as key epigenetic regulators in pancreatic cancer pathology, has not been investigated. This study aims to determine the cytotoxic and apoptotic effects of the combination of GEM and EGCG on PANC1 cells and to examine its effectiveness on the expression levels of microRNAs involved in cancer progression. Material and Method Cytotoxicity of GEM and EGCG in PANC1 cells was assessed using the WST-1 assay, and combination effects were analyzed using isobologram analysis. Apoptosis analysis was performed using the Annexin V method. miRNA isolation was conducted with the miRNeasy Kit, followed by cDNA synthesis using the miScript II Reverse Transcription Kit. Changes in the expression of miRNAs involved in cancer cell proliferation, apoptosis, and metastasis were examined using real-time qRT-PCR analysis. Results The IC50 values for GEM at 24, 48, and 72 hours were determined as 72.85 μM, 26.55 μM, and 9.38 μM, respectively. EGCG's IC50 values at 24, 48, and 72 hours were determined as 64.36 μM, 48.34 μM, and 19.73 μM, respectively. When combined at a 2:3 ratio (GEM: EGCG) at 24 and 72 hours, a synergistic effect was observed, while at 48 hours, a strong synergistic drug interaction was observed. At a concentration of only 26.55 μM, the group treated with GEM showed a 4.2-fold increase in apoptosis compared to the control group receiving fresh medium. In contrast, the combination treatment (EGCG: 4.71 μM, GEM: 3.14 μM) resulted in a remarkable 12.04-fold increase in apoptosis. After combination treatment, the expression of tumor suppressor miRNAs, miR-137, and miR-130a-3p, increased, while the expression of oncogenic miRNAs, including miR-27a-3p, miR-425- 5p, miR-183-5p, miR-187-3p, miR-21-5p, miR-324-5p, and miR-486-5p, decreased. Conclusion EGCG can sensitize pancreatic cancer to GEM through epigenetic mechanisms, shedding light on novel therapeutic approaches. [ABSTRACT FROM AUTHOR]

Purpose: The phosphatidylinositol 3-kinase/AKT/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway is a complex intracellular metabolic pathway that leads to cell growth and tumor proliferation and plays a key role in drug resistance in breast cancer. Therefore, the anti-cancer effects of oleanolic acid (OA), maslinic acid (MA), and their combination were investigated to improve the performance of the treatment strategy. Methods: We investigated the effect of OA and MA on cell viability using the WST-1 method. The synergistic effect of the combination was analyzed by isobologram analysis. In addition, the effects of the two compounds, individually and in combination, on apoptosis, autophagy, and the cell cycle were investigated in MCF7 cells. In addition, changes in the expression of PI3K/AKT/mTOR genes involved in apoptosis, cell cycle and metabolism were determined by quantitative RT-PCR. Results: MA, OA, and a combination of both caused G0/G1 arrest. Apoptosis also increased in all treated groups. The autophagosomal LC3-II formation was induced 1.74-fold in the MAtreated group and 3.25-fold in the MA-OA-treated group. The combination treatment resulted in increased expression of genes such as GSK3B, PTEN, CDKN1B and FOXO3 and decreased expression of IGF1, PRKCB and AKT3 genes. Conclusion: The results showed that the combination of these two substances showed the highest synergistic effect at the lowest dose and using MA-OA caused cancer cells to undergo apoptosis. The use of combination drugs may reduce the resistance of cancer cells to treatment. [ABSTRACT FROM AUTHOR]

Dysregulation of the Wnt signaling pathway contributes to the development of many cancer types. Natural compounds produced with biotechnological systems have been the focus of research for being a new drug candidate both with unlimited resources and cost-effective production. In this study, it was aimed to reveal the effects of isopropylchaetominine on cytotoxic, cytostatic, apoptotic and Wnt signaling pathways in brain, pancreatic and prostate cancer. The IC 50 values of isopropylchaetominine in U-87 MG, PANC1, PC3 and LNCaP cells were calculated as 91.94 μM, 41.68 μM, 54.54 μM and 7.86 μM in 72nd h, respectively. The metabolite arrests the cell cycle in G 0 /G 1 phase in each cancer cells. Iso-propylchaetominine induced a 4.3-fold and 1.9-fold increase in apoptosis in PC3 and PANC1 cells, respectively. The toxicity of isopropylchaetominine in healthy fibroblast cells was assessed using the annexin V method, and no significant apoptotic activity was observed between the groups treated with the active substance and untreated. In U-87 MG, PANC1, PC3, and LNCaP cells under treatment with isopropylchaetominin, the expression levels of DKK3, TLE1, AES, DKK1, FRZB, DAB2, AXIN1/2, PPARD, SFRP4, APC and SOX1 7 tumor suppressor genes increased significantly. Decreases in expression of Wnt1, Wnt2, Wnt3, Wnt4, Wnt5, Wnt6, Wnt10, Wnt11, FRZ2, FRZ3, FRZ7, TCF7L1, BCL9, PYGO, CCND2, c-MYC, WISP1 and CTNNB1 oncogenic genes were detected. All these result shows that isopropylchaetominine can present promising new treatment strategy in different cancer types. [Display omitted] • Iso-propylchaetominine exhibits anticancer activity in in vitro models of brain, pancreatic and prostate cancer. • Iso-propylchaetominin arrests the cell cycle in the G0/G1 phase. • Iso-propylchaetominine regulates the Wnt signaling pathway. • Iso-propylchaetominine causes cell cycle arrest by regulating the Wnt pathway. [ABSTRACT FROM AUTHOR]

Objective(s): STATs are one of the initial targets of emerging anti-cancer agents due to their regulatory roles in survival, apoptosis, drug response, and cellular metabolism in CML. Aberrant STAT3 activity promotes malignancy, and acts as a metabolic switcher in cancer cell metabolism, contributing to resistance to TKI nilotinib. To investigate the possible therapeutic effects of targeting STAT3 to overcome nilotinib resistance by evaluating various cellular responses in both sensitive and nilotinib resistant CML cells and to test the hypothesis that energy metabolism modulation could be a mechanism for re-sensitization to nilotinib in resistant cells. Materials and Methods: By using RNAi-mediated STAT3 gene silencing, cell viability and proliferation assays, apoptotic analysis, expressional regulations of STAT mRNA transcripts, STAT3 total, pTyr705, pSer727 protein expression levels, and metabolic activity as energy metabolism was determined in CML model K562 cells, in vitro. Results: Targeting STAT3 sensitized both parental and especially nilotinib resistant cells by decreasing leukemic cell survival; inducing leukemic cell apoptosis, and decreasing STAT3 mRNA and protein expression levels. Besides, cell energy phenotype was modulated by switching energy metabolism from aerobic glycolysis to mitochondrial respiration in resistant cells. RNAi-mediated STAT3 silencing accelerated the sensitization of leukemia cells to nilotinib treatment, and STAT3-dependent energy metabolism regulation could be another underlying mechanism for regaining nilotinib response. Conclusion: Targeting STAT3 is an efficient strategy for improving the development of novel CML therapeutics for regaining nilotinib response, and re-sensitization of resistant cells could be mediated by induced apoptosis and regulation in energy metabolism. [ABSTRACT FROM AUTHOR]

Photodynamic therapy (PDT) is a safe treatment method for target-specific applications. In this study, the efficacy of PDT on triple-negative breast cancer cells after activation of phthalocyanine derivative CoPc-COOH with RED light was investigated. A combination of CoPc-COOH and 40 J/cm2 showed the highest synergistic effect. [Display omitted] Triple-negative breast cancer (TNBC) has the worst prognosis among breast cancers. The deficiency of hormone receptors (ER-/PR-/HER-2-) limits treatment options. Photodynamic therapy (PDT) is a safe treatment method for target-specific applications and has no toxic side effects. In this study, the efficacy of PDT on TNBC cells after activation of phthalocyanine derivative CoPc-COOH with RED light was investigated. The cytotoxicity of CoPc-COOH in MDA-MB-231 cells was evaluated at 24, 48, and 72 h. To detect phototoxic activity, cells were exposed simultaneously to a dose range of CoPc-COOH at 48th hours and 20, 30, 40, and 50 J/cm2 RED light in a laser source with an output fluorescence of 12 mW/cm2. The apoptotic effect of single CoPc-COOH, RED light, and their combinations were examined by using the Annexin V method and the ROS generation potential using the carboxy-H2-DCFH-DA test in flow cytometry. DCF fluorescence intensity of control and dose groups were taken under the fluorescence microscope. The IC 50 values of CoPc-COOH in MDA-MB-231 cells at the 24th, 48th, and 72nd hours were calculated as 131.04 µM, 66.75 µM, and 59.74 µM, respectively. In the phototoxicity test, 50 J/cm2 application increased the IC 50 dose of CoPc-COOH to 73.73 µM; 20, 30, 40 J/cm2 application reduced the doses to 2.31 µM, 0.20 µM, and 28.14 µM, respectively. Compared to the untreated group, it was determined that ROS formation and apoptotic stimulation were triggered significantly in treatment groups. A combination of CoPc-COOH and 40 J/cm2 showed the highest synergistic effect. [ABSTRACT FROM AUTHOR]

PI3K/Akt/mTOR pathway activation causes relapse and resistance after radiotherapy in breast cancer (BC). We aimed to radiosensitize BC cell lines to irradiation (IR) by PKI-402, a dual PI3K/mTOR inhibitor. We performed cytotoxicity, clonogenicity, hanging drop, apoptosis and double-strand break detection, and phosphorylation of 16 essential proteins involved in the PI3K/mTOR pathway. Our findings showed that PKI-402 has cytotoxic efficiency in all cell lines. Clonogenic assay results showed that PKI-402 plus IR inhibited the colony formation ability of MCF-7 and breast cancer stem cell lines. Results showed that PKI-402 plus IR causes more apoptotic cell death than IR alone in the MCF-7 cells but did not cause significant changes in the MDA-MB-231. γ-H2AX levels were increased in MDA-MB-231 in PKI-402 plus IR groups, whereas we did not observe any apoptotic and γ-H2AX induction in BCSCs and MCF-10A cells in all treatment groups. Some pivotal phosphorylated proteins of the PI3K/AKT pathway decreased, several proteins increased and others did not change. In conclusion, if the combined use of PKI-402 with radiation is supported by in vivo studies, it can contribute to the treatment options and the course of the disease. [ABSTRACT FROM AUTHOR]

MicroRNAs are small RNAs with ability to attach to the large number of RNA that regulate gene expression on post-transcriptional level via inhibition or degradation of specific mRNAs. MiRNAs in cells are the primary regulators of functions such as cell growth, differentiation, and apoptosis and considerably influence cell function. The expression levels of microRNAs change in human diseases, including cancer. These changes highlight their essential role in cancer pathogenesis. Ubiquitous irregular expression profiles of miRNAs have been detected in various human cancers using genome-wide identification techniques, which are emerging as novel diagnostic and prognostic cancer biomarkers of high specificity and sensitivity. The measurable miRNAs with enhanced stability in blood, tissues, and other body fluids provide a comprehensive source of miRNA-dependent biomarkers for human cancers. The leading role of miRNAs as potential biomarkers in human cancers is discussed in this article. In addition, the interests and difficulties of miRNAs as biomarkers have been explored. [ABSTRACT FROM AUTHOR]

A novel gene editing tool, the Cas system, associated with the CRISPR system, is emerging as a potential method for genome modification. This simple method, based on the adaptive immune defense system of prokaryotes, has been developed and used in human cancer research. These technologies have tremendous therapeutic potential, especially in gene therapy, where a patient-specific mutation is genetically corrected to cure diseases that cannot be cured with conventional treatments. However, translating CRISPR/Cas9 into the clinic will be challenging, as we still need to improve the efficiency, specificity, and application of the technology. In this review, we will explain how CRISPR-Cas9 technology can treat cancer at the molecular level, focusing on ordination and the epigenome. We will also focus on the promise and shortcomings of this system to ensure its application in the treatment and prevention of cancer. [ABSTRACT FROM AUTHOR]

Background: Anorexia nervosa (AN) is a serious psychosomatic disorder with unclear pathomechanisms. Metabolic dysregulation is associated with disruption of redox homeostasis that might play a pivotal role in the development of AN. The aim of our study was to assess oxidative status and carbonyl stress in plasma, urine and saliva of patients with AN and healthy controls. Methods: Plasma, spot urine, and saliva were collected from 111 girls with AN (aged from 10 to 18 years) and from 29 age-matched controls. Markers of oxidative stress and antioxidant status were measured using spectrophotometric and fluorometric methods. Results: Plasma advanced oxidation protein products (AOPP) and advanced glycation end products (AGEs) were significantly higher in patients with AN than in healthy controls (by 96, and 82%, respectively). Accordingly, urinary concentrations of AOPP and fructosamines and salivary concentrations of AGEs were higher in girls with AN compared with controls (by 250, and 41% in urine; by 92% in saliva, respectively). Concentrations of thiobarbituric acid reactive substances (TBARS) in saliva were 3-times higher in the patients with AN than in the controls. Overall antioxidants were lower in plasma of girls with AN compared to the controls, as shown by total antioxidant capacity and ratio of reduced and oxidized glutathione (by 43, and 31%, respectively). Conclusions: This is the first study assessing wide range of markers of oxidative status in plasma, urine and saliva of the patients with AN. We showed that both, higher levels of markers of oxidative stress and lower antioxidants play a role in redox disruption. Restoration of redox homeostasis might be of the clinical relevance [ABSTRACT FROM AUTHOR]

Ponatinib is used for advanced treatment of chronic myeloid leukemia (CML), although low doses to prevent side effects do not suppress survival pathways and eradicate leukemia stem cells (LSCs). We evaluated the potential of ponatinib and PI3K/mTOR dual-inhibitor VS-5584 combination (PoVS) therapy to increase the anti-leukemic effects of ponatinib and investigated the underlying mechanisms at the molecular level. We measured the cytotoxicities of ponatinib, VS-5584, and PoVS (CCK-8 assay), and used the median-effect equation for combination analyses. We investigated the effects of inhibitory concentrations on apoptosis, cell viability and cell-cycle regulation (flow cytometry), protein levels (ELISA, Western blot), transcriptional activities (dual-luciferase reporter assay), gene expressions (qRT-PCR). VS-5584 exerted selective cytotoxic effects against CML and LSC cell lines. VS-5584 inhibited the PI3K/Akt/mTOR pathway, resulting in reduced cell viability, slightly induced caspase-independent apoptosis, prominent G 0 /G 1 cell-cycle blockade that is not a consequence of quiescence. Normal hematopoietic stem cell line was the least affected. Moreover, ponatinib and VS-5584 mediated synergistic anti-leukemic effects on leukemic cells. VS-5584 reduced the ponatinib dose required to target leukemic cells. PoVS treatment inhibited PI3K/Akt/mTOR pathway more consistently than either of the two agents alone through reducing p-Akt, p-mTOR, p-S6K, p-PRAS40, p-S6. The subsequent downstream effects were an increase in C/EBP transcriptional activity and decreases in activities of E2F/DP1, Myc/Max, CREB, STAT3, NFκB, AP-1, Elk-1/SRF. Transcriptional regulation resulted in alterations in the expression levels of target mRNAs. Our results highlight PoVS can be a promising treatment strategy for eliminating CML cells and LSCs selectively, with the reduced ponatinib doses. [Display omitted] • Ponatinib and VS-5584 mediate synergistic anti-leukemic effects on leukemic cells. • The combination inhibits PI3K/mTOR pathway stronger than either of the agents alone. • VS-5574 led LSCs to prominent G 0 /G 1 cell-cycle blockade, but not quiescence. • VS-5584 elevates anti-leukemic effects of ponatinib by regulating transcription. • Unlike ponatinib, the combination affects normal hematopoietic stem cells the least. [ABSTRACT FROM AUTHOR]