S3 Cells Research Articles

Characteristics, Chemical Analysis and Biological Activities of Methanol Extracts of Lichens Pleurosticta Acetabulum and Cladonia Subulata

Abstract The aim of this study is to investigate the chemical composition of methanol extracts of the lichens Pleurosticta acetabulum and Cladonia subulata and their antioxidant, and anticancer activities. The phytochemical analysis of the extracts of lichens was determined by HPLC-UV method. The predominant phenolic compounds in these extracts were norstictic acid and salazinic acids in P. acetabulum, while hypoprotocetraric acid and fumarprotocetraric acid were the major metabolites detected in C. subulata. Total phenolics and flavonoids in the extracts were determined spectrophotometrically, with the varied amount from 21.31 to 73.45 mg GA/g and from 8.48 to 15.42 mg RU/g, respectively. The lichen extracts showed comparable and strong antioxidant activity, exhibited higher DPPH and hydroxyl radical scavengings, inhibitory activity towards lipid peroxidation and reducing power. Cytotoxic effects of lichens were tested against HeLa S3 and LS174 cell lines using MTT method.The cytotoxic effects of P. acetabulum and C. subulata extracts toward two cancer cell lines were in the range from 39.17 to >200 μg/mL IC50 value. The present study showed that the tested extracts of lichens demonstrated important antioxidant and anticancer effects. That suggests that these lichens can be used as new sources of the natural antioxidants and anti-cancer compounds.

Comprehensive assessment of peptide derived from pig spleen: Preparation, bioactivity and structure-activity relationships

Pig spleens, as the by-products of pig resources, were often discarded as waste or used for low-value. Therefore, we tried to isolate bioactive peptide from protein hydrolysate of pig spleens to increase their added value. In this study, the low molecular weight peptides (<3000 Da) were prepared by enzyme hydrolysis and its structure-activity relationship was discussed. The main secondary structures of the pig spleen peptide (PSP) were β-turn and random coil by Fourier transform infrared (FT-IR) analysis. Hydrophobic amino acids accounted for half of the total amino acid content. In vitro, PSP had no inhibition effect on HepG2, HCT116, H22 and S180 cells. While PSP exhibited scavenging capacities of PSP on 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide anion (O2−) and hydroxyl (OH) radicals. In vivo, the activity of antioxidant enzymes (superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX)) increased and the level of malondialdehyde (MDA) decreased in PSP-treated mice. PSP displayed obvious immunomodulatory effects including stimulation of multiple immune cells to proliferate, production of cytokines (IFN-γ, IL-2, IL-4 and TNF-α) and increase the level of IgG, IgM and IgA antibodies. It induced solid tumor cells apoptosis and arrested at S phase of cell cycle. In conclusion, PSP had significant immunomodulatory effects and antioxidant activity. These results provided theoretical and data support for bioactive peptides derived from meat by-products used as the functional foods.

Systemic Immunomodulatory Effects of Codonopsis pilosula Glucofructan on S180 Solid-Tumor-Bearing Mice.

The immune functions of the body are intricately intertwined with the onset and advancement of tumors, and immunotherapy mediated by bioactive compounds has exhibited initial effectiveness in overcoming chemotherapy resistance and inhibiting tumor growth. However, the comprehensive interpretation of the roles played by immunologic components in the process of combating tumors remains to be elucidated. In this study, the Codonopsis pilosula glucofructan (CPG) prepared in our previous research was employed as an immunopotentiator, and the impacts of CPG on both the humoral and cellular immunity of S180 tumor-bearing mice were investigated. Results showed that CPG administration of 100 mg/kg could effectively inhibit tumor growth in mice with an inhibitory ratio of 45.37% and significantly improve the expression of Interleukin-2 (IL-2), Interferon-γ (IFN-γ), and Tumor Necrosis Factor-α (TNF-α). Additionally, CPG clearly enhanced B-cell-mediated humoral immunity and immune-cell-mediated cellular immunity, and, finally, induced S180 cell apoptosis by arresting cells in the G0/G1 phase, which might result from the IL-17 signaling pathway. These data may help to improve comprehension surrounding the roles of humoral and cellular immunity in anti-tumor immune responses.

Novel bimetallic oxido-bridged phenolato hafniumIV complex with enhanced anti-tumor activity and aqueous stability

A novel dimeric phenolato Hafnium complex [{HfIV(Salan2,4-Me)(OnBu)}2O] was synthesized by reacting [HfIV(Salan2,4-Me)2] in the presence of 100 equiv. of H2O. The structure was characterized by 1H, 13C NMR spectroscopy, HRMS and X-Ray diffraction. In comparison with [HfIV(Salan2,4-Me)2] and another precursor [{HfIV(Salan2,4-Me)(OnBu)}2O], this dimeric HfIV complex demonstrated enhanced aqueous stability and anti-tumor activity (IC50: 71.2 ± 25.3 μM against Hela S3 cells; 62.2 ± 27.7 μM against Hep G2 cells). Furthermore, this dimeric HfIV complex can only be synthesized from [HfIV(Salan2,4-Me)2] in low equivalent of H2O, upon addition of 2,6-dipicolinic acid, it can be transformed to a Salan/Dipic HfIVbis-chelated complex.

Chemical profile, in vitro bioactivities, and antinociceptive effect of Red Pitaya extracts on zebrafish

Aqueous and ethanolic extracts of red pitaya (Hylocereus polyrhizus) peel, pulp, and seeds were evaluated for their chemical composition by UPLC-Q-TOF-MSE, antioxidant capacity (DPPH and ABTS assays), cytotoxicity and in vitro antiproliferative effect (MTT assay), antinociceptive activity and in vivo toxicity (Zebrafish model). A total of 25 compounds were identified in the extracts, mostly betacyanins, phenolic acids, flavonoids, and organic acids. The seeds extracts displayed both the highest radical scavenging activity and total phenol content. The pulp aqueous extract and ethanolic extracts exhibited some cytotoxicity against Vero cells, depending on the concentration and exposure time. These extracts were also able to inhibit S-180 cells proliferation, however only the peel ethanolic extract showed moderate to high selective indexes (SI: 4.06, 18.63), suggesting that its antiproliferative effect is not associated with the cytotoxic effect towards non-cancer cells, such as Vero cells. Regarding the antinociceptive activity, after adjusting the dose to exclude the sedation effect, the seeds ethanolic extract displayed antinociceptive activity at the inflammatory phase, similar to morphine. The extracts (1.0, 2.5 or 5.0 mg/mL) showed to be safe for zebrafish. In summary, these findings highlight the red pitaya as a promising natural source of bioactive, both in its edible and inedible parts, which can be better probed, for instance as anticancer and antinociceptive agents. In addition, the regular consumption of red pitaya can contribute to the health promotion, and the prevention of chronic diseases.

Design and Synthesis of Poly(2,2'-Bipyridyl) Ligands for Induction of Cell Death in Cancer Cells: Control of Anticancer Activity by Complexation/Decomplexation with Biorelevant Metal Cations.

Chelation therapy is a medical procedure for removing toxic metals from human organs and tissues and for the treatment of diseases by using metal-chelating agents. For example, iron chelation therapy is designed not only for the treatment of metal poisoning but also for some diseases that are induced by iron overload, cancer chemotherapy, and related diseases. However, the use of such metal chelators needs to be generally carried out very carefully, because of the side effects possibly due to the non-specific complexation with intracellular metal cations. Herein, we report on the preparation and characterization of some new poly(bpy) ligands (bpy: 2,2'-bipyridyl) that contain one-three bpy ligand moieties and their anticancer activity against Jurkat, MOLT-4, U937, HeLa S3, and A549 cell lines. The results of MTT assays revealed that the tris(bpy) and bis(bpy) ligands exhibit potent activity for inducing the cell death in cancer cells. Mechanistic studies suggest that the main pathway responsible for the cell death by these poly(bpy) ligands is apoptotic cell death. It was also found that the anticancer activity of the poly(bpy) ligands could be controlled by the complexation (anticancer activity is turned OFF) and decomplexation (anticancer activity is turned ON) with biorelevant metal cations. In this paper, these results will be described.

Living cells as a biological analog of optical tweezers - a non-invasive microrheology approach.

Microrheology, the study of fluids on micron length-scales, promises to reveal insights into cellular biology, including mechanical biomarkers of disease and the interplay between biomechanics and cellular function. Here a minimally-invasive passive microrheology technique is applied to individual living cells by chemically binding a bead to the surface of a cell, and observing the mean squared displacement of the bead at timescales ranging from milliseconds to 100s of seconds. Measurements are repeated over the course of hours, and presented alongside analysis to quantify changes in the cells' low-frequency elastic modulus, G0', and the cell's dynamics over the time window ∼10-2s to 10s. An analogy to optical trapping allows verification of the invariant viscosity of HeLa S3 cells under control conditions and after cytoskeletal disruption. Stiffening of the cell is observed during cytoskeletal rearrangement in the control case, and cell softening when the actin cytoskeleton is disrupted by Latrunculin B. These data correlate with conventional understanding that integrin binding and recruitment triggers cytoskeletal rearrangement. This is, to our knowledge, the first time that cell stiffening has been measured during focal adhesion maturation, and the longest time over which such stiffening has been quantified by any means. STATEMENT OF SIGNIFICANCE: Here, we present an approach for studying mechanical properties of live cells without applying external forces or inserting tracers. Regulation of cellular biomechanics is crucial to healthy cell function. For the first time in literature, we can non-invasively and passively quantify cell mechanics during interactions with functionalised surface. Our method can monitor the maturation of adhesion sites on the surface of individual live cells without disrupting the cell mechanics by applying forces to the cell. We observe a stiffening response in cells over tens of minutes after a bead chemically binds. This stiffening reduces the deformation rate of the cytoskeleton, although the internal force generation increases. Our method has potential for applications to study mechanics during cell-surface and cell-vesicle interactions.

Synergistic Antitumor Effect of Combined Radiotherapy and Engineered Salmonella typhimurium in an Intracranial Sarcoma Mouse Model.

Intracranial sarcoma is an uncommon aggressive cancer with a poor prognosis and a high recurrence rate. Although postoperative adjuvant radiotherapy (RT) is the most recommended treatment strategy, it does not significantly improve survival rates. In this study, we used an attenuated Salmonella typhimurium strain engineered to secrete Vibrio vulnificus flagellin B (SLpFlaB) as an immunotherapy to assist with the antitumor effects of RT on intracranial sarcoma. In vitro, the expression of γH2AX and cleaved caspase-3 was analyzed by Western blot. In vivo detection of SLpFlaB colonization time in tumors was measured using an in vivo imaging system (IVIS). Tumor growth delay and elimination were demonstrated in an intracranial mouse model, and the distribution of macrophages, M1 macrophages, and CD8+ cells after treatment was measured using FACS analysis. Our findings in vitro suggest that combination therapy increases S-180 radiosensitivity, the expression of DNA double-strand breaks, and programmed cell death. In vivo, combination treatment causes intracranial sarcoma to be eliminated without tumor recurrence and redistribution of immune cells in the brain, with data showing the enhanced migration and infiltration of CD8+ T cells and macrophages, and an increased proportion of M1 macrophage polarization. Compared to RT alone, the combination therapy enhanced the radiosensitivity of S-180 cells, promoted the recruitment of immune cells at the tumor site, and prevented tumor recurrence. This combination therapy may provide a new strategy for treating intracranial sarcomas.

Synthesis and cytotoxicity study of water soluble 8-hydroxyquinoline stabilized zirconium(Ⅳ) complexes

Three octacoordinated zirconium(IV) tetrakis-(8-hydroxyquinolinate) complexes [(L1-3)4Zr(IV)] were synthesized smoothly in THF at room temperature using ZrCl4 as a starting material with yields ranging from 74% to 83%. Their structures were characterized by 1H, 13C NMR and X-ray diffraction spectroscopy. [(L1-3)4Zr(IV)] exhibited excellent solubility and stability in H2O and DMSO, and they have demonstrated enhanced cytotoxicity against human Hep G2, Hela S3 and PC9 cells than cisplatin. Among them, [(L1)4Zr(IV)] bearing non-substituted 8-hydroxyquinoline showed about 2 orders of magnitude higher inhibition activity against Hep G2 cells than cisplatin (IC50: 0.14 ± 0.1 μM, cisplatin: 13.82 ± 1.3 μM). The selectivity towards tumor cells was preliminary evidenced by their partial or completely vanished inhibition activity against AML12 cells. Additionally, [(L1)4Zr(IV)] could almost exclusively induce the apoptosis of Hep G2 cells.

Comparative analysis of the occupancy of Histone H3 Lysine 4 methylation in the cells treated with TGFβ and Interferonγ

In this current study, we have compared our H3K4me3 Chip-Sequencing data in PC3 cells in response to 6 h and 24 h TGFβ stimulation with the IFNγ stimulated/unstimulated HeLa S3 cells Since both TGFβ and IFNγ play an essential role in tumorigenesis both as a tumor promoter and tumor suppressor and known to antagonize each other's signalling, it would be of utmost importance to find out the regions undergoing histone modification changes in response to TGFβ and IFNγ and compare them to explore the genes common to both as well as the specific for each ligand. Our study has compared the genes showing H3K4me3 occupancy in response to both TGFβ and IFNγ. Several genes were found to be shared between the TGFβ and IFNγ. DAVID Functional enrichment analysis in the TGFβ and IFNγ dataset revealed association of genes with different biological processes such as miRNA-mediated gene silencing, positive regulation of ERK cascade, hypoxia-induced apoptosis repression, translational regulation and molecular functions such as TGFβR activity, GPCR activity, TGFβ binding activity. Further analysis of these genes can reveal fascinating insights into epigenetic regulation by growth factor stimulation.

Anticancer effects of phytol against Sarcoma (S-180) and Human Leukemic (HL-60) cancer cells.

Phytol (Pyt), a diterpenoid, possesses many important bioactivities. This study evaluates the anticancer effects of Pyt on sarcoma 180 (S-180) and human leukemia (HL-60) cell lines. For this purpose, cells were treated with Pyt (4.72, 7.08, or 14.16 μM) and a cell viability assay was performed. Additionally, the alkaline comet assay and micronucleus test with cytokinesis were also performed using doxorubicin (6 μM) and hydrogen peroxide (10 mM) as positive controls and stressors, respectively. Results revealed that Pyt significantly reduced the viability and rate of division in S-180 and HL-60 cells with IC50 values of 18.98 ± 3.79 and 1.17 ± 0.34 μM, respectively. Pyt at 14.16 μM exerted aneugenic and/or clastogenic effects in S-180 and HL-60 cells, where the number of micronuclei and other nuclear abnormalities (e.g., nucleoplasmic bridges and nuclear buds) were frequently observed. Moreover, Pyt at all concentrations induced apoptosis and showed necrosis at 14.16 μM, suggesting its anticancer effects on the tested cancer cell lines. Taken together, Pyt showed promising anticancer effects, possibly through inducing apoptosis and necrosis mechanisms, and it exerted aneugenic and/or clastogenic effects on the S-180 and HL-60 cell lines.

Triterpenoids of Ganoderma lucidum inhibited S180 sarcoma and H22 hepatoma in mice by regulating gut microbiota

In order to explore effect of natural plant extracts on anti-tumor and prevent tumor development. The study assessed the antitumor effect of triterpenoids of Ganoderma lucidum (TGL) on S180 and H22 tumor bearing mice. A triterpene compound, 2α, 3α, 23-trihydroxy-urs-12-en-28-oic acid, was successfully isolated and purified from G. lucidum. S180 and H22 cells were subcutaneously inoculated in the left axilla of mice to establish a transplantable tumor model. After, the mice were orally treated with TGL and evaluated by tumor inhibition rate, organ index, and the serum index. The Bax and Bcl-2 proteins and gut microbiota was analyzed using western blot and 16S rDNA sequencing respectively. The results showed the tumor inhibition rates of TGL were higher than 40% in H22 and S180 tumor bearing mice. TGL had a protective effect on the spleen and thymus, and improved lipid peroxidation caused by the increased free radicals. TGL downregulated Bcl-2 and upregulated Bax. In particular, TGL treatment improved the reduction of gut microbiota richness and structure.

Lactate: A critical regulator of cell proliferation via anaphase promoting complex remodeling

Lactate: A critical regulator of cell proliferation via anaphase promoting complex remodeling

Chemical modulation of Schistosoma mansoni lysine specific demethylase 1 (SmLSD1) induces wide-scale biological and epigenomic changes.

Background: Schistosoma mansoni, a parasitic worm species responsible for the neglected tropical disease schistosomiasis, undergoes strict developmental regulation of gene expression that is carefully controlled by both genetic and epigenetic processes. As inhibition of S. mansoni epigenetic machinery components impairs key transitions throughout the parasite's digenetic lifecycle, a greater understanding of how epi-drugs affect molecular processes in schistosomes could lead to the development of new anthelmintics. Methods: In vitro whole organism assays were used to assess the anti-schistosomal activity of 39 Homo sapiens Lysine Specific Demethylase 1 (HsLSD1) inhibitors on different parasite life cycle stages. Moreover, tissue-specific stains and genomic analysis shed light on the effect of these small molecules on the parasite biology. Results: Amongst this collection of small molecules, compound 33 was the most potent in reducing ex vivo viabilities of schistosomula, juveniles, miracidia and adults. At its sub-lethal concentration to adults (3.13 µM), compound 33 also significantly impacted oviposition, ovarian as well as vitellarian architecture and gonadal/neoblast stem cell proliferation. ATAC-seq analysis of adults demonstrated that compound 33 significantly affected chromatin structure (intragenic regions > intergenic regions), especially in genes differentially expressed in cell populations (e.g., germinal stem cells, hes2 + stem cell progeny, S1 cells and late female germinal cells) associated with these ex vivo phenotypes. KEGG analyses further highlighted that chromatin structure of genes associated with sugar metabolism as well as TGF-beta and Wnt signalling were also significantly perturbed by compound 33 treatment. Conclusions: This work confirms the importance of histone methylation in S. mansoni lifecycle transitions, suggesting that evaluation of LSD1 - targeting epi-drugs may facilitate the search for next-generation anti-schistosomal drugs. The ability of compound 33 to modulate chromatin structure as well as inhibit parasite survival, oviposition and stem cell proliferation warrants further investigations of this compound and its epigenetic target SmLSD1.

Schistosoma mansoni coactivator associated arginine methyltransferase 1 (SmCARM1) effect on parasite reproduction.

The human blood fluke parasite Schistosoma mansoni relies on diverse mechanisms to adapt to its diverse environments and hosts. Epigenetic mechanisms play a central role in gene expression regulation, culminating in such adaptations. Protein arginine methyltransferases (PRMTs) promote posttranslational modifications, modulating the function of histones and non-histone targets. The coactivator-associated arginine methyltransferase 1 (CARM1/PRMT4) is one of the S. mansoni proteins with the PRMT core domain. We carried out in silico analyses to verify the expression of SmPRMTs in public datasets from different infection stages, single-sex versus mixed-worms, and cell types. The SmCARM1 function was evaluated by RNA interference. Gene expression levels were assessed, and phenotypic alterations were analyzed in vitro, in vivo, and ex vivo. The scRNAseq data showed that SmPRMTs expression is not enriched in any cell cluster in adult worms or schistosomula, except for Smcarm1 expression which is enriched in clusters of ambiguous cells and Smprmt1 in NDF+ neurons and stem/germinal cells from schistosomula. Smprmt1 is also enriched in S1 and late female germ cells from adult worms. After dsRNA exposure in vitro, we observed a Smcarm1 knockdown in schistosomula and adult worms, 83 and 69%, respectively. Smcarm1-knockdown resulted in reduced oviposition and no significant changes in the schistosomula or adult worm phenotypes. In vivo analysis after murine infection with Smcarm1 knocked-down schistosomula, showed no significant change in the number of worms recovered from mice, however, a significant reduction in the number of eggs recovered was detected. The ex vivo worms presented a significant decrease in the ovary area with a lower degree of cell differentiation, vitelline glands cell disorganization, and a decrease in the testicular lobe area. The worm tegument presented a lower number of tubercles, and the ventral sucker of the parasites presented a damaged tegument and points of detachment from the parasite body. This work brings the first functional characterization of SmCARM1 shedding light on its roles in S. mansoni biology and its potential as a drug target. Additional studies are necessary to investigate whether the reported effects of Smcarm1 knockdown are a consequence of the SmCARM1-mediated methylation of histone tails involved in DNA packaging or other non-histone proteins.

Development of Wireless Power-Transmission-Based Photodynamic Therapy for the Induction of Cell Death in Cancer Cells by Cyclometalated Iridium(III) Complexes.

Photodynamic therapy (PDT), a noninvasive method for cancer therapy, involves the generation of reactive oxygen species (ROS) by the photochemical excitation of photosensitizers (PSs) to induce cell death in cancer cells. A variety of PS including porphyrin derivatives and metal complexes such as iridium (Ir) complexes have been reported. In clinical trials, red-near infrared (NIR) light (650-900 nm) is preferred for the excitation of PSs due to its deeper penetration into tissues compared with visible light (400-500 nm). To overcome this limitation, we established a PDT system that uses cyclometalated iridium(III) (Ir(III)) complexes that are excited with blue light in the wireless power transmission (WPT) system. To achieve this, we developed a light-emitting diode (LED) light device equipped with a receiver coil that receives electricity from the transmitter coil through magnetic resonance coupling. The LEDs in the receiving device use blue light (470 nm) to irradiate a given Ir(III) complex and excite triplet oxygen (3O2) to singlet oxygen (1O2) which induces cell death in HeLa S3 cells (human cervical carcinoma cells). The results obtained in this study suggest that WPT-based PDT represents a potentially new method for the treatment of tumors by a non-battery LED, which are otherwise difficult to treat by previous PDT systems.

Characterization of the human &lt;i&gt;IDH1&lt;/i&gt; gene promoter

&lt;abstract&gt; &lt;p&gt;In cancer, the production of ATP depends mainly on glycolysis, usually accompanied by the dysfunction of the tricarboxylic acid (TCA) cycle and oxidative phosphorylation (OXPHOS). Nicotinamide adenine dinucleotide (NAD&lt;sup&gt;+&lt;/sup&gt;) is a coenzyme for various biological enzymatic reactions such as those involved in the TCA cycle. To investigate the molecular mechanisms involved in carcinogenesis, the transcription system of genes associated with mitochondrial function should be elucidated. In this study, we isolated several mitochondrial function-associated bidirectional promoters and tested whether they responded to NAD&lt;sup&gt;+&lt;/sup&gt;-metabolism regulating compounds, namely, &lt;italic&gt;trans&lt;/italic&gt;-resveratrol (Rsv), 2-deoxy-D-glucose (2DG), 3-amino benzamide (3AB), and olaparib (OLA), in HeLa S3 cells. Transient transfection and luciferase (Luc) reporter assay showed that the &lt;italic&gt;IDH1&lt;/italic&gt; promoter was prominently activated by these compounds. The &lt;italic&gt;IDH1&lt;/italic&gt; gene, which encodes a nicotinamide adenine dinucleotide phosphate (NADP&lt;sup&gt;+&lt;/sup&gt;) dependent isocitrate dehydrogenase, is frequently mutated in glioma and leukemia cells. In this study, RT-PCR showed that &lt;italic&gt;IDH1&lt;/italic&gt; gene and protein expression was induced in response to the NAD&lt;sup&gt;+&lt;/sup&gt;-regulating drugs Rsv and 3AB. However, IDH1 protein amount was rather stable at control level. The result suggested that a post-transcriptional controlling system works to keep IDH1 at a stable level.&lt;/p&gt; &lt;/abstract&gt;

Over-expression of miR-183-5p or miR-492 triggers invasion and proliferation and loss of polarity in non-neoplastic breast epithelium

microRNAs (miRNAs) serve as novel noninvasive cancer biomarkers. In an HMT-3522 S1 (S1) breast epithelial risk-progression three-dimensional (3D) culture model, non-neoplastic S1 cells form a fully polarized epithelium. When silenced for the gap junction and tumor suppressor Cx43, Cx43-KO-S1 cells recapitulate pre-neoplastic phenotypes observed in tissues at risk for breast cancer in vivo. To delineate the role of miRNAs in breast tumorigenesis and identify key miRNA players in breast epithelial polarity, the miRNA profile specific to Cx43 loss in Cx43-KO-S1 compared to S1 cells was sequenced, revealing 65 differentially expressed miRNAs. A comparative analysis was conducted between these miRNAs and tumor-associated miRNAs from a young Lebanese patient validation cohort. miR-183-5p, downstream of Cx43 loss, was commonly upregulated in the patient cohort and the 3D culture model. miR-492, not attributed to Cx43 loss, was only specifically up-regulated in the young Lebanese patients. Ectopic expression of either miR-183-5p or miR-492 in S1 cells, through pLenti-III-miR-GPF vectors, resulted in the formation of larger multi-layered acini devoid of lumen, with disrupted epithelial polarity, as shown by an altered localization of Cx43, ß-catenin and Scrib, and decreased nuclear circularity in 3D cultures. Enhanced proliferation and invasion capacity were also observed. Over-expression of miR-183-5p or miR-492, therefore, induces pre-neoplastic phenotypes similar to those reported upon Cx43 loss, and may act as oncomiRs and possible biomarkers of increased breast cancer risk.

Proteome-Wide Identification of RNA-Dependent Proteins in Lung Cancer Cells.

Following the concept of RNA dependence and exploiting its application in the R-DeeP screening approach, we have identified RNA-dependent proteins in A549 lung adenocarcinoma cells. RNA-dependent proteins are defined as proteins whose interactome depends on RNA and thus entails RNA-binding proteins (RBPs) as well as proteins in ribonucleoprotein complexes (RNPs) without direct RNA interaction. With this proteome-wide technique based on sucrose density gradient ultracentrifugation and fractionation followed by quantitative mass spectrometry and bioinformatic analysis, we have identified 1189 RNA-dependent proteins including 170 proteins which had never been linked to RNA before. R-DeeP provides quantitative information on the fraction of a protein being RNA-dependent as well as it allows the reconstruction of protein complexes based on co-segregation. The RNA dependence of three newly identified RNA-dependent proteins, DOCK5, ELMO2, also known as CED12A, and ABRAXAS1, also known as CCDC98, was validated using western blot analysis, and the direct RNA interaction was verified by iCLIP2 for the migration-related protein DOCK5 and the mitosis-related protein ABRAXAS1. The R-DeeP 2.0 database provides proteome-wide and cell line-specific information from A549 and HeLa S3 cells on proteins and their RNA dependence to contribute to understanding the functional role of RNA and RNA-binding proteins in cancer cells.

Synthesis and X-ray structure analysis of cytotoxic heptacoordinated Salan hafnium(IV) complexes stabilized with 2,6-dipicolinic acid

Four novel Salan Hf(IV) complexes stabilized by 2,6-dipicolinic acid (Dipic) were synthesized and characterized by 1H, 13C NMR and X-ray diffraction spectroscopy. These Hf(IV)bis-chelates could be obtained in good to excellent yields (88%–91%) and demonstrated rather good stability in aqueous media and on silica gel. [L2Hf(IV)Dipic4-H,Cl] containing steric bulk L2 were stable in about 10% H2O (H2O/THF (v/v)), however, [L1Hf(IV)Dipic4-H,Cl] with non-steric L1 could slowly dissociate and release nontoxic L1. [L1–2Hf(IV)Dipic4-Cl] showed excellent anti-tumoral activity in the range of cisplatin (Hela S3: IC50 = 3.5 ± 0.4 μM, Hep G2: IC50 = 11.2 ± 2.1 μM). In addition, the cellular uptake and apoptosis investigation of [L1Hf(IV)Dipic4-Cl] suggested a fast cellular uptake process against Hela S3 cells with an almost exclusive induced apoptosis cell death path.