Lidocaine Inhibits Rat Prostate Cancer Cell Invasiveness and Voltage-Gated Sodium Channel Expression in Plasma Membrane.

There is increasing evidence, mostly from breast cancer, that use of local anaesthetics during surgery can inhibit disease recurrence by suppressing the motility of the cancer cells dependent on inherent voltage-gated sodium channels (VGSCs). Here, the possibility that lidocaine could affect cellular behaviours associated with metastasis was tested using the Dunning cell model of rat prostate cancer. Mostly, the strongly metastatic (VGSC-expressing) Mat-LyLu cells were used under both normoxic and hypoxic conditions. The weakly metastatic AT-2 cells served for comparison in some experiments. Lidocaine (1-500 µM) had no effect on cell viability or growth but suppressed Matrigel invasion dose dependently in both normoxia and hypoxia. Used as a control, tetrodotoxin produced similar effects. Exposure to hypoxia increased Nav1.7 mRNA expression but VGSCα protein level in plasma membrane was reduced. Lidocaine under both normoxia and hypoxia had no effect on Nav1.7 mRNA expression. VGSCα protein expression was suppressed by lidocaine under normoxia but no effect was seen in hypoxia. It is concluded that lidocaine can suppress prostate cancer invasiveness without effecting cellular growth or viability. Extended to the clinic, the results would suggest that use of lidocaine, and possibly other local anaesthetics, during surgery can suppress any tendency for post-operative progression of prostate cancer.

Prediction of Cell Migration in MDA-MB 231 and MCF-7 Human Breast Cancer Cells Treated with Albizia Lebbeck Methanolic Extract Using Multilinear Regression and Artificial Intelligence-Based Models.

Breast cancer is a common cancer affecting women worldwide, and it progresses from breast tissue to other parts of the body through a process called metastasis. Albizia lebbeck is a valuable plant with medicinal properties due to some active biological macromolecules, and it's cultivated in subtropical and tropical regions of the world. This study reports the phytochemical compositions, the cytotoxic, anti-proliferative and anti-migratory potential of A. lebbeck methanolic (ALM) extract on strongly and weakly metastatic MDA-MB 231 and MCF-7 human breast cancer cells, respectively. Furthermore, we employed and compared an artificial neural network (ANN), an adaptive neuro-fuzzy inference system (ANFIS), and multilinear regression analysis (MLR) to predict cell migration on the treated cancer cells with various concentrations of the extract using our experimental data. Lower concentrations of the ALM extract (10, 5 & 2.5 µg/mL) showed no significant effect. Higher concentrations (25, 50, 100 & 200 µg/mL) revealed a significant effect on the cytotoxicity and proliferation of the cells when compared with the untreated group (p < 0.05; n ≥ 3). Furthermore, the extract revealed a significant decrease in the motility index of the cells with increased extract concentrations (p < 0.05; n ≥ 3). The comparative study of the models observed that both the classical linear MLR and AI-based models could predict metastasis in MDA-MB 231 and MCF-7 cells. Overall, various ALM extract concentrations showed promising an-metastatic potential in both cells, with increased concentration and incubation period. The outcomes of MLR and AI-based models on our data revealed the best performance. They will provide future development in assessing the anti-migratory efficacies of medicinal plants in breast cancer metastasis.

Deep neural network to differentiate brain activity between patients with euthymic bipolar disorders and healthy controls during verbal fluency performance: A multichannel near-infrared spectroscopy study.

In this study, we aimed to differentiate between euthymic bipolar disorder (BD) patients and healthy controls (HC) based on frontal activity measured by fNIRS that were converted to spectrograms with Convolutional Neural Networks (CNN). And also, we investigated brain regions that cause this distinction. In total, 29 BD patients and 28 HCs were recruited. Their brain cortical activities were measured using fNIRS while performing letter versions of VFT. Each one of the 24 fNIRS channels was converted to a 2D spectrogram on which a CNN architecture was designed and utilized for classification. We found that our CNN algorithm using fNIRS activity during a VFT is able to differentiate subjects with BD from healthy controls with 90% accuracy, 80% sensitivity, and 100% specificity. Moreover, validation performance reached an AUC of 94%. From our individual channel analyses, we observed channels corresponding to the left inferior frontal gyrus (left-IFC), medial frontal cortex (MFC), right dorsolateral prefrontal cortex (DLPFC), Broca area, and right premotor have considerable activity variation to distinguish patients from HC. fNIRS activity during VFT can be used as a potential marker to classify euthymic BD patients from HCs. Activity particularly in the MFC, left-IFC, Broca's area, and DLPFC have a considerable variation to distinguish patients from healthy controls.

Mechanisms of Drug Resistance and Use of Nanoparticle Delivery to Overcome Resistance in Breast Cancers.

Breast cancer is the leading cancer type diagnosed among women in the world. Unfortunately, drug resistance to current breast cancer chemotherapeutics remains the main challenge for a higher survival rate. The recent progress in the nanoparticle platforms and distinct features of nanoparticles that enhance the efficacy of therapeutic agents, such as improved delivery efficacy, increased intracellular cytotoxicity, and reduced side effects, hold great promise to overcome the observed drug resistance. Currently, multifaceted investigations are probing the resistance mechanisms associated with clinical drugs, and identifying new breast cancer-associated molecular targets that may lead to improved therapeutic approaches with the nanoparticle platforms. Nanoparticle platforms including siRNA, antibody-specific targeting and the role of nanoparticles in cellular processes and their effect on breast cancer were discussed in this article.

Biosynthesized ZnO Nanoparticles Using Albizia lebbeck Extract Induced Biochemical and Morphological Alterations in Wistar Rats.

Nano-based particles synthesized via green routes have a particular structure that is useful in biomedical applications as they provide cheap, eco-friendly, and non-toxic nanoparticles. In the present study, we reported the effect of various concentrations of Zinc oxide nanoparticles synthesized using A. lebbeck stem bark extract (ZnO NPsAL) as stabilizing agent on rat biochemical profiles and tissue morphology. Adult Wistar rats weighing 170 ± 5 g were randomly classified into eight groups of five rats each; Group A served as a control fed with normal diet and water. Groups B1, B2, C1, C2, D1, D2, and E were treated with 40 mg/kg and 80 mg/kg of the 0.01, 0.05, and 0.1 M biosynthesized ZnO NPsAL and zinc nitrate daily by the gavage method, respectively. The rats were anesthetized 24 h after the last treatment, blood samples, kidney, heart, and liver tissues were collected for biochemical and histopathological analysis. The rats mean body weight, serum alkaline phosphatase, alanine aminotransferase, creatinine, urea, bilirubin, protein, albumin, globulin, total cholesterol, triacylglycerol, and high-density lipoprotein were significantly altered with an increased concentration of biosynthesized ZnO NPsAL when compared with the control group (p < 0.05; n ≥ 5). Furthermore, histopathological analysis of treated rats' kidney, heart, and liver tissue revealed vascular congestion, tubular necrosis, inflammation, and cytoplasmic vacuolation. Biosynthesized ZnO NPsAL showed significant alteration in biochemical parameters and tissue morphology in rats with increasing concentrations of the nanoparticles.

Riluzole: Anti-invasive effects on rat prostate cancer cells under normoxic and hypoxic conditions.

Anti-invasive effects of riluzole and ranolazine, a neuro-protectant and an anti-anginal drug, respectively, on Mat-LyLu rat prostate cancer (PCa) cells were tested in vitro (a) at non-toxic doses and (b) under both normoxic and hypoxic conditions, the latter common to growing tumours. Tetrodotoxin (TTX) was used as a positive control. Hypoxia had no effect on cell viability but reduced growth at 48 hours. Riluzole (5 µmol/L) or ranolazine (20 µmol/L) had no effect on cell viability or growth under normoxia or hypoxia over 24 hours. Matrigel invasion was not affected by hypoxia but inhibited by TTX, ranolazine and riluzole under a range of conditions. The expression of Nav1.7 mRNA, the prevailing, pro-invasive voltage-gated sodium channel α-subunit (VGSCα), was up-regulated by hypoxia. Riluzole had no effect on Nav1.7 mRNA expression in normoxia but significantly reduced it in hypoxia. VGSCα protein expression in plasma membrane was reduced in hypoxia; riluzole increased it but only under hypoxia. It was concluded (a) that riluzole and ranolazine have anti-invasive effects on rat PCa cells and (b) that Nav1.7 mRNA and protein expression can be modulated by riluzole under hypoxia. Overall, therefore, riluzole and ranolazine may ultimately be "repurposed" as anti-metastatic drugs against PCa.

Biosynthesis of zinc oxide nanoparticles using Albizia lebbeck stem bark, and evaluation of its antimicrobial, antioxidant, and cytotoxic activities on human breast cancer cell lines.

BACKGROUND: Biocompatibility and stability of zinc oxide nanoparticles (ZnO NPs) synthesized using plants is an interesting research area of study in nanotechnology, due to its wide applications in biomedical, industrial, cell imaging, and biosensor fields. The present study reports the novel green synthesis of stable ZnO NPs using various concentrations of zinc nitrate (0.01M, 0.05M, 0.1M) and Albizia lebbeck stem bark extracts as an efficient chelating agent. Antimicrobial, antioxidant, cytotoxic, and antiproliferative activities of the synthesized NPs on human breast cancer cell lines were evaluated using different assays. METHODS: Characterization of the synthesized ZnO NPs were carried out using various spectroscopic and microscopic techniques. Antimicrobial activity evaluation using disc diffusion method, antioxidant activity using hydrogen peroxide (H2O2) free radical scavenging assay and cytotoxic activity on MDA-MB 231 and MCF-7 using tryphan blue dye exclusion and MTT assay. RESULTS: The UV-vis spectroscopy result revealed an absorption peak in the range of 370 nm. The involvements of A. lebbeck bioactive compounds in the stabilization of the ZnO NPs were confirmed by X-ray diffraction and Fourier transform infrared analysis. Zeta sizer studies showed an average size of 66.25 nm with a polydisparity index of 0.262. Scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX) analyses results revealed irregular spherical morphology and the presence of primarily Zn, C, O, Na, P, and K, respectively. The biosynthesized ZnO NPs revealed strong antimicrobial potentials against various gram-negative and gram-positive bacterial pathogens. Antioxidant activities carried out using H2O2 free radical scavenging assay revealed higher IC50 values of 48.5, 48.7, and 60.2 µg/mL for 0.1M, 0.05M, and 0.01M ZnO NPs, respectively. Moreover, the biosynthesized ZnO NPs showed significant cytotoxic effects on MDA-MB 231 and MCF-7 breast cancer cell lines (P< 0.001, n≥3) in a concentration-dependent manner. CONCLUSION: Overall, various concentrations of ZnO NPs were synthesized through a stable, simple, and eco-friendly green route via the use of A. lebbeck stem bark extract. The biosynthesized ZnO NPs showed strong antimicrobial, antioxidant and cytotoxic activity against strongly and weakly metastatic breast cancer cell lines.

Anti-proliferative and cytotoxic activities of Allium autumnale P. H. Davis (Amaryllidaceae) on human breast cancer cell lines MCF-7 and MDA-MB-231.

BACKGROUND: Natural products obtained from plants can be potent sources for developing a variety of pharmaceutical products. Allium species have been widely studied for their anti-cancer effects and presented promising results as potential anti-cancer agents. Breast cancer (BCa) is one of the most commonly diagnosed types of cancer in women. In this study, we aimed to investigate the anti-proliferative, cytotoxic and anti-metastatic effects of bulb and stem extracts from Allium autumnale P. H. Davis (Amaryllidaceae), an endemic Allium species to the island of Cyprus, in a comparative approach to weakly metastatic MCF-7 and strongly metastatic MDA-MB-231 breast cancer (BCa) cell lines. METHODS: Possible cytotoxic, anti-proliferative and anti-metastatic effects of the Allium extracts on MCF-7 and MDA-MB-231 cells were tested using trypan blue exclusion, MTT and wound heal assays, respectively. Gas Chromatography Mass Spectroscopy (GC-MS) analysis was performed to determine the prominent medically important compounds in Allium autumnale bulb (AAB) and Allium autumnale stem (AAS) extracts. Student unpaired t-test or ANOVA followed by Newman-Keuls post hoc analysis (INSTAT Software) was used where appropriate. RESULTS: Our results demonstrate that AAB extract (24, 48 and 72 h) exerts significant anti-proliferative effect on both MCF-7 and MDA-MB-231 cells where this effect for AAS extract was observed only at high (5000 and 10,000 µg/mL) concentrations. Cell viability experiments revealed that AAB extract incubations caused more cytotoxicity on both BCa cell lines compared to the AAS. In contrast, there was no effect on lateral motilities of either cell line. CONCLUSION: Overall, our studies demonstrated the anti-cancer activities associated with Allium autumnale, revealing it's cytotoxic and anti-proliferative potential to be further utilized in in vivo studies.

Intracellular calcium oscillations in strongly metastatic human breast and prostate cancer cells: control by voltage-gated sodium channel activity.

The possible association of intracellular Ca2+ with metastasis in human cancer cells is poorly understood. We have studied Ca2+ signaling in human prostate and breast cancer cell lines of strongly versus weakly metastatic potential in a comparative approach. Intracellular free Ca2+ was measured using a membrane-permeant fluorescent Ca2+-indicator dye (Fluo-4 AM) and confocal microscopy. Spontaneous Ca2+ oscillations were observed in a proportion of strongly metastatic human prostate and breast cancer cells (PC-3M and MDA-MB-231, respectively). In contrast, no such oscillations were observed in weakly/non metastatic LNCaP and MCF-7 cells, although a rise in the resting Ca2+ level could be induced by applying a high-K+ solution. Various parameters of the oscillations depended on extracellular Ca2+ and voltage-gated Na+ channel activity. Treatment with either tetrodotoxin (a general blocker of voltage-gated Na+ channels) or ranolazine (a blocker of the persistent component of the channel current) suppressed the Ca2+ oscillations. It is concluded that the functional voltage-gated Na+ channel expression in strongly metastatic cancer cells makes a significant contribution to generation of oscillatory intracellular Ca2+ activity. Possible mechanisms and consequences of the Ca2+ oscillations are discussed.

Oxidative stress causes plasma protein modification.

We investigated the effect of oxidative systems on plasma proteins using Chloramine-T, a source of free radicals. Plasma specimens from 10 healthy volunteers were treated with 40 mmol/L Chloramine-T (1:1 v/v). Total protein and plasma carbonyl levels were evaluated spectrophotometrically. Identification of plasma proteins modifications was performed by SDS-PAGE, protein and lipid electrophoresis. Protein fragmentation was evaluated by HPLC. Total protein levels of oxidised plasmas were significantly lower (4.08 ± 0.12 g/dL) than control (7.86 ± 0.03 g/dL) (P < 0.01). Plasma carbonyl levels were higher (1.94 ± 0.38 nmol/mg protein) in oxidised plasma than that of control (0.03 ± 0.01 nmol/mg protein) (P < 0.01). Plasma oxidation had no significant effect on the levels of proteins and lipids. Protein fragmentations were detected in oxidised groups compared to those of the control. We conclude that protein modifications have direct effect on the protein functions, which are related to stress agent, its treatment period(s), and the methodology used for evaluating such experimental results.

Molecular origin of plasma membrane citrate transporter in human prostate epithelial cells.

The prostate is a highly specialized mammalian organ that produces and releases large amounts of citrate. However, the citrate release mechanism is not known. Here, we present the results of molecular cloning of a citrate transporter from human normal prostate epithelial PNT2-C2 cells shown previously to express such a mechanism. By using rapid amplification of cDNA ends PCR, we determined that the prostatic carrier is an isoform of the mitochondrial transporter SLC25A1 with a different first exon. We confirmed the functionality of the clone by expressing it in human embryonic kidney cells and performing radiotracer experiments and whole-cell patch-clamp recordings. By using short interfering RNAs targeting different parts of the sequence, we confirmed that the cloned protein is the main prostatic transporter responsible for citrate release. We also produced a specific antibody and localized the cloned transporter protein to the plasma membrane of the cells. By using the same antibody, we have shown that the cloned transporter is expressed in non-malignant human tissues.

Citrate transport and metabolism in mammalian cells: prostate epithelial cells and prostate cancer.

Citrate, an organic trivalent anion, is a major substrate for generation of energy in most cells. It is produced in mitochondria and used either in the Krebs' cycle or released into cytoplasm through a specific mitochondrial carriers. Citrate can also be taken up from blood through different plasma membrane transporters. In the cytoplasm, citrate can be used ultimately for fatty acid synthesis, which is increased in cancer cells. Here, we review the ways in which citrate can be transported and discuss the changes in transport and metabolism that occur in cancer cells. The primary focus is on the prostate gland, which is known to produce and release large amounts of citrate during its normal secretory function. The significant changes that occur in citrate-related metabolism and transport in prostate cancer are the second focus. This review strives to relate these mechanisms to molecular biology on the one hand and to clinical applications on the other.