WRL-68 Cells Research Articles

Development of Cistanche deserticola Fermented Juice and Its Repair Effect on Ethanol-Induced WRL68 Cell Damage

Cistanche deserticola is a valuable Chinese herb, but traditional dry processing causes the loss of active substances. This study developed Cistanche deserticola fermented juice (CFJ) using lactic acid bacteria and optimized the fermentation process to achieve the maximum active substance content and taste. More interestingly, superoxide dismutase (SOD) activity was increased during fermentation, and CFJ exerted a reparative effect on ethanol-induced cell damage. SOD activity reached 603.26 U/mL when the ratios in the total inoculum volume of Lactobacillus reuteri, Lactococcus pentosus, Streptococcus thermophilus, Bifidobacterium animalis, Lactobacillus casei, and Lactobacillus acidophilus were 31.74%, 15.71%, 17.45%, 11.65%, 9.56%, and 13.89%, respectively. Further, the optimal fermentation conditions for CFJ were determined using a response surface methodology. More importantly, CFJ promoted the proliferation of WRL68 cells, and CFJ exerted an obvious reparative effect on ethanol-treated cells, in which the cell survival rate increased to 120.35 ± 0.77% (p < 0.05). The underlying mechanism might have been that CFJ reduced the MDA content in damaged cells from 1.36 nmol/mg prot to 0.88 nmol/mg prot and increased GSH-Px and SOD activities by 48% and 72%, respectively. This study provides a theoretical basis and reference data for the fermentation of C. deserticola and its hepatoprotective activity.

Pharmacognostic Profile and Screening of Anti-Proliferative Potential of Methanolic Extract of Tripterygium wilfordii Plant on WRL-68 Cell Line and Function of Polycystin-1.

Tripterygium wilfordii is a medicinal plant that plays a crucial role in health care programs, especially in developing countries, and had anti-tumor, anti-inflammatory, anti-fertility, anti-bacterial, and other therapeutic effects. This study was designed to determine the anti-proliferative effects of methanolic extract of T. wilfordii on the WRL-68 cell line and the function of polycystin-1 (PC-1). The half-maximal inhibitory concentration (IC50) values were recorded in WRL-68 and AsPC-1 cell lines as 193 µg/ml and 149.2 µg/ml, respectively, at 2-2.55 and 2-2.2 µg/ml methanolic plant concentrations. The maximum cytotoxic activities of the extract on the growth inhibition of WRL-68 and AsPC-1 were generally observed at 97.64% and 95.94% at extract concentrations of 50 µg/ml and 25 µg/ml, respectively. The pharmacognostic profile of T. wilfordii extract was found to be alkaloids, tannins, terpenoides, flavonoids, glycosides, and phenols. The extracts of T. wilfordii were tested through gas chromatography-mass spectrometry showing four peaks representing mostly of 3-Oxobutanol; ethyl acetate; acetic acid ethyl ester; chlorbromuron; 1-(methylthio)-, (E)-; n-Hexadecanoic acid; tetradecanoic acid; and 9-Octadecenoic acid. Therefore, the results of this study revealed that the methanolic extract of T. wilfordii was more potential in inducing anti-proliferative activity of WRL-68 and AsPC-1 human cell lines than the control. In addition, the current study was the first study that reported the anti-proliferative potential of T. wilfordii in the treatment of human embryonic liver WRL-68 cancer cells.

Molecular characterization of wild Pleurotus ostreatus (MW457626) and evaluation of β-glucans polysaccharide activities

Pleurotus ostreatus is a common cultivated edible mushroom worldwide. The fruiting bodies of P. ostreatus is a rich source of a $\beta$-glucans polysaccharide. The current study aimed to investigate the effectiveness of $\beta$-glucans as a natural polysaccharide produced by P. ostreatus as an antioxidant, antimicrobial, and anticancer. The molecular identification of P. ostreatus isolate was confirmed by Internal Transcribed Spacer (ITS) sequence. The sequence alignment and phylogenetic evolutionary relationship of studied ITS sequence were performed against some deposited sequences in GenBank. The analysis of high-performance liquid chromatography (HPLC) as well as the result of fourier transform infrared spectroscopy (FTIR) has confirmed the presence of $\beta$-glucans polysaccharide in the tested samples. The percentage of antioxidant activity of $\beta$-glucans showed a gradual increase from 8.59% to 12.36, 18.56, 23.69, 44.66 and 80.36% at the concentrations of 31.2, 64.4, 125, 250, 500, and 800 $\mu$g/ml, respectively. In addition, all concentrations of $\beta$-glucans showed higher antioxidant activities when compared with standard antioxidant (Vitamin C). The highest antimicrobial activity of $\beta$-glucans polysaccharide was against P. aeruginosa with a zone of inhibition (45 mm), while the lowest activity was against S. aureus (13 mm) both at 100 mg/mL. The percentage of growth-inhibiting of MCF-7 a humanbreast cancer cell line and normal WRL-68 cell line affected by $\beta$-glucans were determined by 3-(4,5)-dimethylthiazol (-z-y1)-3,5-di-phenytetrazoliumromide (MTT assay).

Synthesis of New 1,2,4-Triazole Derivatives with Expected Biological Activities

To further study novel N-aceyl, N-thiourea and imidazole derivatives were synthesized. We used a multi-step reaction protocol that began with symmetrical 4-amino- 1,2,4-Triazole [I]. The new Schiff base [II] was derived by condensation of compound [I] with 4-aminoacetophenone in Ethanol and some drops from glacial acetic acid as a catalyst, whereas the Schiff base [III] was synthesized by reacting compound [II] with 4-hydroxyacetophenone in ethanol. N- acyl derivative [IV] was prepared by addition reaction of acetyl chloride to azomethine group of Schiff base [III] in dry benzene. Then, N-thiourea derivative [V] was prepared from reaction of thiourea with N-aceyl derivative [IV] in alkaline solution. The third step involved cyclization reaction of derivative [V] with 2-hydroxy-1,2-diphenylethan-1-one in dimethyl formamide (DMF) to obtain new imidazole derivative [VI]. The synthesized compounds were characterized on the spectroscopic data and their physical properties. Also, we studied the anti-bacterial activity of the prepared compounds against three types of bacteria: Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa, some of which showed best antibacterial activity comparable with Ampicillin, as standard antibiotic. Furthermore, the cytotoxic effect of various concentrations of the prepared compounds [II] and [VI] was investigated against MCF-7 (human breast carcinoma cells), Hep G2 (human liver cancer cell line) and WRL-68 (human hepatic cell line), revealing a moderate activity at 400 μl/ml, which had no effect on the growth of normal WRL-68 cells, and confirmed the safety of using this type of molecules in medications.

Hormetic effect of low doses of rapamycin triggers anti-aging cascades in WRL-68 cells by modulating an mTOR-mitochondria cross-talk.

Rapamycin is hormetic in nature-it demonstrates contrasting effects at high and low doses. It is toxic at moderate/high doses, while it can restrain aging and extend lifespan at low doses. However, it is not fully understood how rapamycin governs cellular aging. On the other hand, aging is putatively correlated to mitochondrial dysregulation. Although previous studies have suggested that hormetic (low) doses of rapamycin can cause partial/incomplete inhibition of mTOR, the actual modus operandi of how such partial mTOR inhibition might modulate the mTOR-mitochondria cross-talk remained to be deciphered in the context of cellular aging. The present study was designed to understand the hormetic effects of rapamycin on cellular factors that govern aging-associated changes in mitochondrial facets, such as functional and metabolic homeostases, sustenance of membrane potential, biogenesis, mitophagy, and oxidative injury to mitochondrial macromolecules. WRL-68 cells treated (24h) with variable doses of rapamycin were studied for estimating their viability, apoptosis, senescence, mitochondrial density and Δψm. Expression levels of key functional proteins were estimated by immunofluorescence/immunoblots. Oxidative damage to mtDNA/mtRNA/proteins was measured in mitochondrial lysates. We demonstrated that hormetic doses (0.1 and 1nM) of rapamycin can alleviate aging-associated mitochondrial dyshomeostasis in WRL-68 cells, such as oxidative injury to mitochondrial nucleic acids and proteins, as well as disequilibrium of mitochondrial density, membrane potential, biogenesis, mitophagy and overall metabolism. We established that low doses of rapamycin can hormetically amend the mTOR-mitochondria cross-talk, and can consequently promote anti-aging outcome in cells.

Physicochemical Properties, Antioxidant Activity and Protective Effect on Oxidative Stress Cells of Vernonia amygdalina Delile Leaf Jiaosu during Fermentation

Vernonia amygdalina Delile leaf was used as raw material to prepare Jiaosu. The change of basic physicochemical indexes were monitored during fermentation, and the antioxidant capacity was determined by the scavenging ability against ABTS+ radical and hydroxyl radical, and its protective effect to human normal hepatocytes (WRL-68) on oxidative stress was investigated by cell experiment. Results: Compared with the jiaosu on day 2 and day 55, the pH decreased from 4.24±0.01 to 3.46±0.07, and the content of soluble protein, total sugar, total phenol and total flavonoids decreased from 0.95±0.02, 244.94±4.89, 6.05±0.03, 0.37±0.01 mg/mL to 0.71±0.02, 34.26±0.57, 3.42±0.21 and 0.08±0.01 mg/mL, respectively. The total acid content increased from 0.15±0.01 to 0.95±0.01 mg/mL. The content of ascorbic acid increased from 25.36±0.52 μg/ml to 28.44±0.01 μg/mL from 2 to 28 days, and then decreased. The ABTS+ radical scavenging ability was the strongest on the 18th day 73.72%±0.86%, and the hydroxyl radical scavenging capacity was the highest on the 28th day 54.14%±0.43%. There was a significant positive correlation between ABTS+ radical, hydroxyl radical scavenging ability and total acid content (P<0.01). After H2O2-injured WRL-68 cell, in cellular ROS was significant decreased, and the activities of antioxidant enzymes SOD, CAT and GSH-Px were increased in the Jiaosu treated group. In summary, the Jiaosu of Vernonia amygdalina Delile leaf had strong ability to scavenge free radicals in the middle stage of fermentation, and the ability of WRL-68 cells to resist oxidative stress was improved after pretreatment with it.

FIRST REPORT OF THE ANTICANCER EFFICIENCY OF AGARICUS BRASILIENSIS MUSHROOM ON HUMAN EMBRYONIC LIVER WRL68 AND HUMAN PANCREATIC ASPC-1 CANCER CELLS THERAPY

Mushrooms are food traditionally consumed in Asia, Europe and America. They are being studied for medicinal benefits. Extensive studies have shown that Agaricus brasiliensis mushroom used as a medical productto combat cancers. Our data reveal that the determined inhibitory concentration fifty(IC50) values were observed maximum dose responses (IC50) of WRL68 andAsPC-1 cancer cells reported of 172.6 µg/ml and 158.2 µg/ml respectively at 2.23 and 2.1 µg/ml ethanolic mushroom concentrations. The highly cytotoxic activity of the extract on growth inhibition AsPC-1 and WRL68were generally observed 97.9% and 95% at extract concentrations of 25 µg/ml and 50µg/ml respectively. Finally, Phytochemical profile of Agaricus brasiliensis mushroom extract found to beflavonoids, glycosides, saponins, phenols,alkaloids, tannins.The extracts of Agaricus brasiliensiswas tested throughGas Chromatography-Mass(GC-MS). There were five different compounds analyzed from the extracts of Agaricus brasiliensis. The compounds in the ethanolic extract of A. brasiliensis mushroom were comprised mostly of Acetic acid ethyl ester(38.39%),followed by(3-Methyl-oxiran-2-yl)-methanol (34.71%), Chlorbromuron (22.86%), n-Hexadecanoic acid (3.85%) and Heptane, 1-(1-butenyloxy) (0.19%). However, no studies were done using Agaricus brasiliensismushroom extract against WRL68 and AsPC-1 human cell lines. Therefore, the anticancer efficiency of Agaricus brasiliensis mushroom on human embryonic liver WRL68 andhuman pancreatic AsPC-1 cancer cellstherapy is being reported for the first time in the current study.

Dexmedetomidine hydrochloride inhibits hepatocyte apoptosis and inflammation by activating the lncRNA TUG1/miR-194/SIRT1 signaling pathway

BackgroundLiver injury seriously threatens the health of people. Meanwhile, dexmedetomidine hydrochloride (DEX) can protect against liver injury. However, the mechanism by which Dex mediates the progression of liver injury remains unclear. Thus, this study aimed to investigate the function of DEX in oxygen and glucose deprivation (OGD)-treated hepatocytes and its underlying mechanism.MethodsIn order to investigate the function of DEX in liver injury, WRL-68 cells were treated with OGD. Cell viability was measured by MTT assay. Cell apoptosis was detected by flow cytometry. Inflammatory cytokines levels were measured by ELISA assay. The interaction between miR-194 and TUG1 or SIRT1 was detected by dual-luciferase reporter. Gene and protein levels were measured by qPCR or western blotting.ResultsDEX notably reversed OGD-induced inflammation and apoptosis in WRL-68 cell. Meanwhile, the effect of OGD on TUG1, SIRT1 and miR-194 expression in WRL-68 cells was reversed by DEX treatment. However, TUG1 knockdown or miR-194 overexpression reversed the function of DEX in OGD-treated WRL-68 cells. Moreover, TUG1 could promote the expression of SIRT1 by sponging miR-194. Furthermore, knockdown of TUG1 promoted OGD-induced cell growth inhibition and inflammatory responses, while miR-194 inhibitor or SIRT1 overexpression partially reversed this phenomenon.ConclusionsDEX could suppress OGD-induced hepatocyte apoptosis and inflammation by mediation of TUG1/miR-194/SIRT1 axis. Therefore, this study might provide a scientific basis for the application of DEX on liver injury treatment.

Intermittent hypoxia preconditioning protects WRL68 cells against oxidative injury: Involvement of the PINK1/Parkin-mediated mitophagy regulated by nuclear respiratory factor 1

The protective effect of intermittent hypoxia (IH) preconditioning against oxidative injury in hepatic cells was investigated and the involvement of the PINK1/Parkin-mediated mitophagy regulated by nuclear respiratory factor 1 (NRF-1) was evaluated. The results showed that IH preconditioning protected HepG2 cells against oxygen and glucose deprivation/reperfusion (OGD/Rep)-induced injury and protected WRL68 cells against H2O2 or AMA-induced oxidative injury. IH preconditioning up-regulated the protein level of NRF-1, PINK1, Parkin, and LC3 II, promoted the recruitment of the cytosolic Parkin, indicating the initiation of the PINK1/Parkin-mediated mitophagy in WRL68 cells. When NRF-1 was down-regulated by NRF-1 specific shRNA, the protein level of PINK1 and Parkin as well as the mitophagy level were significantly decreased. After IH preconditioning, the protein level of PINK1 and the recruitment of Parkin in CCCP-treated group were significantly higher than that of the control group, indicating the increased mitophagy capacity. And the increased mitophagy capacity induced by IH preconditioning was also reduced by down-regulation of NRF-1. Furthermore, the protective effect of IH preconditioning against H2O2-induced oxidative injury in WRL68 cells was inhibited when NRF-1 or PINK1 was down-regulated by specific shRNA. Mitochondrial ROS generation may be responsible for the increased expression of NRF-1 induced by IH preconditioning. In conclusion, the PINK1/Parkin-mediated mitophagy regulated by NRF-1 was involved in IH preconditioning-induced protective effect against oxidative cellular injury in hepatic cells.

SOD mimetic cerium oxide nanorods protect human hepatocytes from oxidative stress

Recently, cerium oxide nanoparticles (CeNPs) have emerged as potential materials for several biomedical applications due to their excellent biocompatibility and biological enzyme-like activities. The mechanism of observed antioxidant (superoxide dismutase and catalase) enzyme-like activities of spherical-shaped CeNPs is suggested due to the switching between its two oxidation states (3+ and 4+). The shape of nanoparticles plays a significant role in controlling the catalytic activity; therefore, we have studied the antioxidant enzyme-like activities of cerium oxide nanorods. Our results suggest that CeNPs (rods) display excellent SOD-like activity as well as auto regeneration ability of its surface “Ce” atoms, comparable to CeNPs (spheres). The cytotoxicity and genotoxicity studies of CeNPs (rods) showed non-toxicity to WRL-68 (human hepatocyte) cells and impart protection from the harmful effects of reactive oxygen species generated in the presence of Buthionine sulfoximine (BSO). BSO exposure induces inhibition of glutathione (GSH) synthesis that regulates the redox status of the cytosol. The cellular internalization of CeNPs (rods) in WRL-68 cells revealed quick uptake by the cells in a time and concentration-dependent manner. The surface regeneration property of CeNPs (rods) was analysed in the presence of hydrogen peroxide (H2O2). Results suggested that the atoms present on the surface of CeNPs (rods) were oxidised into “4+” states after the addition of H2O2. After incubation of 15 days, they regained their original “3+” oxidation state by regenerating their surface, similar to spherical nanoparticles.

WITHDRAWN: Cytotoxicity, genotoxicity and efficacy evaluation of the quercus infectoria in HepG2 and WRL68 cell lines

This article has been withdrawn: please see Elsevier Policy on Article Withdrawal ( https://www.elsevier.com/about/our-business/policies/article-withdrawal ). This article has been withdrawn as part of the withdrawal of the Proceedings of the International Conference on Emerging Trends in Materials Science, Technology and Engineering (ICMSTE2K21). Subsequent to acceptance of these Proceedings papers by the responsible Guest Editors, Dr S. Sakthivel, Dr S. Karthikeyan and Dr I. A. Palani, several serious concerns arose regarding the integrity and veracity of the conference organisation and peer-review process. After a thorough investigation, the peer-review process was confirmed to fall beneath the high standards expected by Materials Today: Proceedings. The veracity of the conference also remains subject to serious doubt and therefore the entire Proceedings has been withdrawn in order to correct the scholarly record.

Am Synthesis, Docking Study and In vitro Anticancer Evaluation of Some New Flurbiprofen Derivatives Against MCF-7 and WRL-68 Cell Lines

A new series of flurbiprofen derivatives containing thiosemicarbazide moiety (3-7) was synthesized from flurbiprofen as parent nucleus by esterification, hydrazide formation, and heating with different aryl isothiocyanate substituents, respectively. Flurbiprofen was also treated with thiosemicarbazide in the presence POCl3 as a catalyst, to produce 1,3,4 -thiadiazole -2-amine (8). Treatment of (8) with different aryl isothiocyanates produced thiourea derivatives (9-12). Also, the reaction of (8) with different benzoyl chloride substituents produced benzamide compounds (13-15). Eventually , treatment of (8) with ethyl acetoacetate(EAA) produced [1,3,4]thiadiazolo[3,2-a]pyrimidin-7-one (16) .The new compounds were characterized by spectroscopic techniques :FTIR, 1HNMR, and CHNS analysis. A molecular docking study for the synthesized compounds (3-16), against the Vascular Endothelial Growth Factor receptor (VEGFR-2) was applied and it indicated that compounds 4,7,13, and 15,exhibited the optimum binding energy of -6.77, -6.12,-6.68, and -6.43 kcal/mol, respectively. Target compounds were also assessed for their in vitro anticancer effects in a cell-line study. All of the compounds tested showed the most plausible anticancer activity, compared to a positive control(Sorafenib), using in vitro MTT cytotoxic assay ,against human breast tumor (MCF-7), and normal WRL-68 cell line. The in vitro results revealed that compounds 4,5,10,11,13, and 15 exhibited the highest inhibitory activity at their IC50 concentrations, against MCF-7 cell lines, as follows (122.7,113.9,95,7. 109.1,40.32 and 112.29µg/mL, respectively. While their cytotoxic effect against normal WRL-68 cell line at their IC50 concentrations, as follow 210.2, 181.3 ,151.7,278.7,80.28, and 236 µg/mL, respectively, therefore, such compounds were considered more selective toward MCF-7 than normal WRL-68,and their selectivity index (SI): 1.71,1.59,1.59 ,2.55 ,1.99 , and 2.10,respectively . Among the synthesized compounds, the compound 15 was chosen to screen its effect in vitro through multi-parameter cytotoxic assay against MCF-7 breast cancer implemented in High Content Screening (ArrayScan XTI, Thermo Scientific),which could be taken in consideration as a starting point for the development of new anticancer drugs

Study the effect of purified alginate from Pseudomonas aeruginosa on Hep G2 and A549 cell lines.

Alginate was extracted from highest alginate producer of P. aeruginosa 29 isolate, it was determined bybiochemical method (Carbazole-borate assay) and molecular method (PCR), where the amplified algD genesamples showed that clear bands at one level for all the produced isolates and molecular weight were 313 onthe leader scale. partially purified of alginate using gel filtration chromatography (Sephadex G-200) showedthe present of single peak and initial alginate fraction at 17 to last fraction 28 of alginate concentration ,where the highest alginate concentration was at fraction 23 . The cytotoxic effect of different concentrationsof purified alginate on cancer cells (HePG2 , A549 ) and normal cells WRL68 was tested. The result ofhalf-maximal inhibition concentration (IC 50) values of purified alginate treated WRL68 and HepG2 cellsafter 24,48,72 hours of incubation at 37o C were ( 187.8 , 62.0, 83.07) ?g /ml , (102.3, 42.5, 61.03) ?g/mlas respectively. While, the IC50 of purified alginate treated WRL68 and A549 cells after 24,48,72 hours ofincubation at 37oC were ( 126.2, 63.57, 90.31 ) ?g /ml , (117.8, 57.06, 65.7) ?g /ml as respectively.

Phloridzin docosahexaenoate, a novel polyphenolic derivative, is cytotoxic to canine osteosarcoma D17 cells

Canine osteosarcoma (OSA) is the most common form of bone cancer diagnosticated in dogs and is highly metastatic. There has been limited advancement in discovering an effective treatment for OSA in the last few decades. The major drawback of the currently used chemotherapeutic drugs is their side effects. In this preliminary study, we investigated the efficacy of using a novel food-derived drug, phloridzin docosahexaenoate (PZ-DHA), in the treatment of canine OSA in vitro. PZ-DHA was selectively cytotoxic to canine OSA D17 cells, while normal human liver cells (WRL68) were more resistant. We also found that PZ-DHA had enhanced cellular uptake in D17 cells compared to its precursors and in WRL68 cell line. This study provides preliminary evidence that PZ-DHA needs to be further assessed as a safe and efficacious new drug in the treatment of both canine and human OSA.

Synthesis, Docking Study And In Vitro Anticancer Evaluation Of New Derivatives Of 2-(1-(2-Flouro-[1,1-Biphenyl]-4-Yl)Ethyl)-6-(Substituted Phenyl) Imidazole[2,1-B][1,3,4]Thiadiazole Derived From Flurbiprofen

Objectives: New derivatives of 2-(1-(2-flouro-[1,1-biphenyl]-4-yl)ethyl)-6-(substituted phenyl) imidazole[2,1-b][1,3,4]thiadiazole, compounds(2-5), were designed and synthesized from the starting parent molecule, flurbiprofen. Materials and Methods: Compound (1) was synthesized by refluxing flurbiprofen, as a starting material , with phosphorus oxychloride (POCl3), and thiosemicarbazide, resulting in flurbiprofen -1,3,4 thiadiazole -2 amine derivative. The target compounds(2-5) were obtained by refluxing compound(1) with different substituted phenacyl bromides, Results: The new compounds were characterized by FTIR, 1HNMRs and CHNS analysis. A molecular docking study of the synthesized compounds (2-5) against HIV protease indicated such compounds occupied the critical site of HIV protease pocket, and demonstrated excellent positioning of the compounds in the pocket. The best binding energy values were (-7.49, -7.22 , -7.51 and-7.12 kcal/mol) for the compounds (2 -5(, respectively. The anticipated physio-chemical parameters(calculated logarithmic of partition cofficent(Clog p),molecular volume (MV), number of violations (nviol), topological surface area (TPSA), and percent of absorption(%Abs.) were computed using software Molinspiration. In vitro outcomes, the target compounds were assessed by cell line study for their anticancer effects. All the tested compounds showed the most plausible anticancer activity, when compared to a positive control (atazanavir), using MTT cytotoxic assay, against human prostatic tumor (PC-3), glioma cell line (LN68), and normal WRL-68 cell line. Conclusion: The results revealed that compounds 2,3,4 and 5 exhibited the highest inhibitory activity against PC3 cell line at IC50 concentrations of 178.2, 75.09, 129.3and 110.5μg/mL, respectively, and they had moderate effects against LN299 cell line at IC50 concentrations 203,172.5, 169,and 157.7 μg/mL, respectively, compared to atazanavir, as a positive control against PC-3, at IC50, 157.3 μg/mL, and in LN299, at IC50 195.7 μg/mL. Also compound (5) attributed to cell-cycle arrest, and induction of apoptosis. The target compounds could be considered as promising as potential anticancer drugs.

PARP-1 activation after oxidative insult promotes energy stress-dependent phosphorylation of YAP1 and reduces cell viability.

Poly(ADP-ribose) polymerase 1 (PARP-1) is a nuclear enzyme that catalyze the transfer of ADP-ribose units from NAD+ to several target proteins involved in cellular stress responses. Using WRL68 (HeLa derivate) cells, we previously showed that PARP-1 activation induced by oxidative stress after H2O2 treatment lead to depletion of cellular NAD+ and ATP, which promoted cell death. In this work, LC-MS/MS-based phosphoproteomics in WRL68 cells showed that the oxidative damage induced by H2O2 increased the phosphorylation of YAP1, a transcriptional co-activator involved in cell survival, and modified the phosphorylation of other proteins involved in transcription. Genetic or pharmacological inhibition of PARP-1 in H2O2-treated cells reduced YAP1 phosphorylation and degradation and increased cell viability. YAP1 silencing abrogated the protective effect of PARP-1 inhibition, indicating that YAP1 is important for the survival of WRL68 cells exposed to oxidative damage. Supplementation of NAD+ also reduced YAP1 phosphorylation, suggesting that the loss of cellular NAD+ caused by PARP-1 activation after oxidative treatment is responsible for the phosphorylation of YAP1. Finally, PARP-1 silencing after oxidative treatment diminished the activation of the metabolic sensor AMPK. Since NAD+ supplementation reduced the phosphorylation of some AMPK substrates, we hypothesized that the loss of cellular NAD+ after PARP-1 activation may induce an energy stress that activates AMPK. In summary, we showed a new crucial role of PARP-1 in the response to oxidative stress in which PARP-1 activation reduced cell viability by promoting the phosphorylation and degradation of YAP1 through a mechanism that involves the depletion of NAD+.

Synthesis, characterization and biocompatibility studies of carbon quantum dots from Phoenix dactylifera.

In the present study, Carbon Quantum Dots (CQDs) were synthesized from Phoenix dactylifera (Date palm fruit) using microwave-assisted pyrolysis and were characterized for its various properties. The synthesized CQD sample exhibited a narrow absorbance peak at 270nm in UV-Vis spectrum that indicated generation of narrow sized particles. The FTIR analysis of the crude CQDs and dialysed sample revealed the various functional groups involved in the formation of CQDs. TEM data revealed the nature of CQDs to be quasi-spherical and spatially distributed. Biocompatibility of the CQDs was studied using various model systems. CQDs displayed no cytotoxic and anti-clonogenic property when exposed to WRL-68 cell line whereas a slight toxicity was evident in HT1080 post 24h of incubation suggesting the tremendous potential of the CQDs in the synergistic killing of cancer cells. Phytotoxicity assessment in four different seedlings revealed the non-toxic nature of CQDs. Further these CQDs were found to possess high biocompatibility imposing no inhibition in microbial growth and zilch effect on the development of zebrafish embryos. Thus these CQDs can find immense potential applications in fields of biomedicine as biomolecule detection, drug carriers, fluorescent tracers and in controlling the drug release.

A novel indenone derivative selectively induces senescence in MDA-MB-231 (breast adenocarcinoma) cells

Triple-negative breast cancer is the most aggressive form of breast cancer with limited intervention options. Moreover, a number of belligerent therapeutic strategies adopted to treat such aggressive forms of cancer have demonstrated detrimental side effects. This necessitates exploration of targeted chemotherapeutics. We assessed the efficacy of a novel indenone derivative (nID) [(±)-N-(2-(-5-methoxy-1-oxo-3-(2-oxo-2-phenylethyl)-2,3-dihydro-1H-inden-2-yl)ethyl)-4-methylbenzenesulfonamide], synthesized by a novel internal nucleophile-assisted palladium-catalyzed hydration-olefin insertion cascade; against triple-negative breast cancer cells (MDA-MB-231). On 24 h treatment, the nID caused decline in the viability of MDA-MB-231 and MDA-MB-468 cells, but did not significantly (P < 0.05) affect WRL-68 (epithelial-like) cells. In fact, the nID demonstrated augmentation of p53 expression, and consequent p53-dependent senescence in both MDA-MB-231 and MDA-MB-468 cells, but not in WRL-68 cells. The breast cancer cells also exhibited reduced proliferation, downregulated p65/NF-κB and survivin, along with augmented p21Cip1/WAF1 expression, on treatment with the nID. This ensued cell cycle arrest at G1 stage, which might have driven the MDA-MB-231 cells to senescence. We observed a selectivity of the nID to target MDA-MB-231 cells, whereas WRL-68 cells did not show any considerable effect. The results underscored that the nID has potential to be developed into a cancer therapeutic.

Antiproliferative and Pro-Apoptotic Effect of Uvaol in Human Hepatocarcinoma HepG2 Cells by Affecting G0/G1 Cell Cycle Arrest, ROS Production and AKT/PI3K Signaling Pathway

Natural products have a significant role in the development of new drugs, being relevant the pentacyclic triterpenes extracted from Olea europaea L. Anticancer effect of uvaol, a natural triterpene, has been scarcely studied. The aim of this study was to understand the anticancer mechanism of uvaol in the HepG2 cell line. Cytotoxicity results showed a selectivity effect of uvaol with higher influence in HepG2 than WRL68 cells used as control. Our results show that uvaol has a clear and selective anticancer activity in HepG2 cells supported by a significant anti-migratory capacity and a significant increase in the expression of HSP-60. Furthermore, the administration of this triterpene induces cell arrest in the G0/G1 phase, as well as an increase in the rate of cell apoptosis. These results are supported by a decrease in the expression of the anti-apoptotic protein Bcl2, an increase in the expression of the pro-apoptotic protein Bax, together with a down-regulation of the AKT/PI3K signaling pathway. A reduction in reactive oxygen species (ROS) levels in HepG2 cells was also observed. Altogether, results showed anti-proliferative and pro-apoptotic effect of uvaol on hepatocellular carcinoma, constituting an interesting challenge in the development of new treatments against this type of cancer.

Bovine serum albumin decorated gold nanoclusters: A fluorescence-based nanoprobe for detection of intracellular hydrogen peroxide

Gold nanoclusters are well studied due to their facile synthesis, biocompatibility, limited photobleaching, and bright fluorescence properties. Monitoring the reactive oxygen species (ROS) level in mammalian cells is required to assess the healthy metabolism of oxygen, cell signaling, and homeostasis. ROS are also known to cause several human diseases including cancer, neurodegenerative disease, diabetes, and cardiovascular disorders, therefore, following the ROS status could provide important information about the critical pathophysiology of the diseased cells/tissues. Among different ROS, hydrogen peroxide is a well-reported stable oxidative stress biomarker for several diseases. In this work, we report the synthesis of BSA coated AuNCs (BSA-AuNCs) displaying bright red fluorescence (Ex./Em. 511/651 nm), which utilized for the degradation of hydrogen peroxide with concomitant loss of fluorescence of BSA-AuNCs. The fluorescence property of this nanoprobe was used for the detection of hydrogen peroxide levels in liver cells (WRL-68). The accumulation of hydrogen peroxide in WRL-68 cells was artificially induced by 3-amino-1,2,4-triazole (catalase inhibitor) treatment, which leads to a decrease in the fluorescence intensity of BSA-AuNCs to enable fluorescence-based sensing of hydrogen peroxide. Thus, the synthesized fluorescent BSA-AuNCs could be used for the detection of intracellular hydrogen peroxide.