Bone Cancer Cell Lines Research Articles

Effect of Regiochemistry and Methylation on the Anticancer Activity of a Ferrocene-Containing Organometallic Nucleoside Analogue.

Four new bis-substituted ferrocene derivatives containing either a hydroxyalkyl or methoxyalkyl group and either a thyminyl or methylthyminyl group have been synthesised and characterised by a range of spectroscopic and analytical techniques. They were included in a structure-activity-relationship (SAR) study probing anticancer activities in osteosarcoma (bone cancer) cell lines and were compared with a known lead compound, 1-(S,Rp ), a nucleoside analogue that is highly toxic to cancer cells. Biological studies using the MTT assay revealed that a regioisomer of ferronucleoside 1-(S,Rp ), which only differs from the lead compound in being substituted on two cyclopentadienyl rings rather than one, was over 20 times less cytotoxic. On the other hand, methylated derivatives of 1-(S,Rp ) showed comparable cytotoxicities to the lead compound. Overall these studies indicate that a mechanism of action for 1-(S,Rp ) cannot proceed through alcohol phosphorylation and that its geometry and size, rather than any particular functional group, are crucial factors in explaining its high anticancer activity.

Comparison of methods for the isolation of cell-free DNA from cell culture supernatant.

In vitro characterization of cell-free DNA using two-dimensional cell culture models is emerging as an important step toward an improved understanding of the physical and biological characteristics of cell-free DNA in human biology. However, precise measurement of the cell-free DNA in cell culture medium is highly dependent on the efficacy of the method used for DNA purification, and is often a juncture of experimental confusion. Therefore, in this study, we compared six commercially available cell-free DNA isolation kits for the recovery of cell-free DNA from the cell culture supernatant of a human bone cancer cell line (143B), including two magnetic bead-based manual kits, one automated magnetic bead-based extraction method, and three manual spin-column kits. Based on cell-free DNA quantitation and sizing, using the Qubit dsDNA HS assay and Bioanalyzer HS DNA assay, respectively, the different methods showed significant variability concerning recovery, reproducibility, and size discrimination. These findings highlight the importance of selecting a cell-free DNA extraction method that is appropriate for the aims of a study. For example, mutational analysis of cell-free DNA may be enhanced by a method that favors a high yield or is biased toward the isolation of short cell-free DNA fragments. In contrast, quantitative analysis of cell-free DNA in a comparative setting (e.g. measuring the fluctuation of cell-free DNA levels over time) may require the selection of a cell-free DNA isolation method that forgoes a high recovery for high reproducibility and minimal size bias.

Salvador‐Warts‐Hippo Genes Methylation status in Osteosarcoma Cell Lines

Osteosarcoma is an aggressive malignant bone tumour greatly prevalent in children and adolescent. The Salvador‐Warts‐Hippo signalling pathway is a highly conserved and is important in regulating organ size, cell proliferation and cell death. Tumorigenesis has been extensively studied along with the genetic alterations of the promoters of Salvador‐Warts‐Hippo genes which promote poor prognosis. Although, the epigenetical modifications, which present a fundamental controlling points for genetic regulations, remain inadequately clear. Such as the methylation status of CpG islands which are associated to key components genes of the hippo signalling pathway which includes MST1, MST2, LATS1, LATS2, WW45 and SAV1. We applied two pairs of primers were needed with one pair specific for methylated DNA (M) and the other for unmethylated DNA in several bone cancer cell lines including SaOS2, HT1080 and Ewing’s sarcoma A673. Combined Bisulfite Restriction Analysis (CoBRA) carried out on these cell lines and the methylation patterns were varied between osteosarcoma and Ewing’s sarcoma cells. This approach of combination techniques provides a clear note of the methylation status profile of CpG islands that governs the expression of these Salvador‐Warts‐Hippo genes in osteosarcoma cell lines. In addition, the level of expression of Salvador‐Warts‐Hippo pathway genes during demethylating agents treatment play a corresponding evidence of the biological relevance that the methylation status has on the transcriptome. The outcomes of the present study offer distinguishable means that allow the ability to differentiate between several epigenetic variations in different osteosarcoma cell lines. We conclude that Salvador‐Warts‐Hippo needs further investigation in osteosarcoma progression.Support or Funding InformationDeanship of graduate studies, King Saud University, Riyadh, Saudi Arabia

Alendronic Acid as Ionic Liquid: New Perspective on Osteosarcoma.

Herein the quantitative synthesis of eight new mono- and dianionic Organic Salts and Ionic Liquids (OSILs) from alendronic acid (ALN) is reported by following two distinct sustainable and straightforward methodologies, according to the type of cation. The prepared ALN-OSILs were characterized by spectroscopic techniques and their solubility in water and biological fluids was determined. An evaluation of the toxicity towards human healthy cells and also human breast, lung and bone (osteosarcoma) cell lines was performed. Globally, it was observed that the monoanionic OSILs showed lower toxicity than the corresponding dianionic structures to all cell types. The highest cytotoxic effect was observed in OSILs containing a [C2OHMIM] cation, in particular [C2OHMIM][ALN]. The latter showed an improvement in IC50 values of ca. three orders of magnitude for the lung and bone cancer cell lines as well as fibroblasts in comparison with ALN. The development of OSILs with high cytotoxicity effect towards the tested cancer cell types, and containing an anti-resorbing molecule such as ALN may represent a promising strategy for the development of new pharmacological tools to be used in those pathological conditions.

Osteogenic silver oxide doped mesoporous bioactive glass for controlled release of doxorubicin against bone cancer cell line (MG-63): In vitro and in vivo cytotoxicity evaluation

Mesoporous bioactive glass demonstrated valuable biomedical applications especially in the field of bone regeneration and drug delivery. For the treatment of cancer by chemotherapy, controlled drug delivery system is one of the significant methods. In this work, silver oxide doped mesoporous bioactive glass nanoparticles (Ag2O-MBG NPs) were developed for the controlled release of doxorubicin as a model drug. Ag2O-MBG NPs were prepared by using the sol-gel method and employing pluronic 123 as an internal template. The prepared Ag2O-MBG NPs were characterized by energy-dispersive X-ray spectroscopy (EDX), scanning electron microscopy (SEM) and transmission electron microscopy (TEM) for composition, shape, morphology, and size. Surface area and pore size were determined by the Brunauer-Emmett-Teller (BET) and Barrett-Joyner-Halenda (BJH) method respectively. When immersed in simulated body fluid (SBF), hydroxycarbonate apatite (HCAp) formation was demonstrated as established by Fourier Transform Infrared (FTIR) spectroscopy and X-ray diffraction (XRD). The as-synthesized Ag2O-MBG NPs did not show any detrimental effects during MTT assay and in vivo tissue histopathology. Doxorubicin (DOX) was encapsulated with an efficiency of 84% and its release was observed to be affected by the drug loading concentration (0.2–1.0 mg/mL) and pH (6.4–8.4) of the release media. DOX release was of 93% was witnessed approximately for two weeks at a slight acidic pH of 6.4. At 11.88 μg/mL of DOX-Ag2O-MBG NPs, notable inhibitory effects on the viability of the MG-63 osteosarcoma cancer cells were observed. These features proved that the Ag2O-MBG NPs system is effective for bone tissue regeneration and bone cancer treatment.

Design of a Thiosemicarbazide-Functionalized Calix[4]arene Ligand and Related Transition Metal Complexes: Synthesis, Characterization, and Biological Studies.

In this study, we synthesized a new thiosemicarbazide-functionalized calix[4]arene L and its Co2+, Ni2+, Cu2+, and Zn2+ transition metal complexes. For characterization several techniques were employed: Fourier-transform infrared (FT-IR), 1H nuclear magnetic resonance (NMR), 13C-NMR, 15N-NMR, correlation spectroscopy (COZY), nuclear Overhauser enhancement spectroscopy (NOESY), electrospray ionization (ESI)–mass spectroscopy, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and elemental analysis. To explore the capability of the thiosemicarbazide function hosted on a calix[4]arene scaffold for growth inhibition of bacteria, fungi, and cancerous tumor cells, a series of biological evaluations were performed. For L, the antimicrobial tests revealed a higher antibacterial activity against gram-positive Bacillus subtilis and a lower activity against gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa), whereas the gram-positive Staphylococcus aureus shows resistance. All examined metal derivatives show an enhancement of the antibacterial activity against gram-negative E. coli bacteria, with a more significant improvement for the Ni2+ and Zn2+ complexes. MTT assays showed a considerable in vitro anticancer activity of Co2+, Ni2+, and Cu2+ complexes against Saos-2 bone cancer cell lines. The activity is ascribable to the inorganic ions rather than calixarene ligand. Hemolysis assay results demonstrated that all compounds have high blood compatibility.

Synthesis, Crystal Structure, Anti-Bone Cancer Activity and Molecular Docking Investigations of the Heterocyclic Compound 1-((2S,3S)-2-(Benzyloxy)Pentan-3-yl) -4-(4-(4-(4-Hydroxyphenyl)Piperazin-1-yl) Phenyl)-1H-1,2,4-Triazol-5(4H)-One

Heterocyclic compound 1-((2S,3S)-2-(benzyloxy)pentan-3-yl)-4-(4-(4-(4-hydroxyphenyl)piperazin-1-yl)phenyl)-1H-1,2,4-triazol-5(4H)-one (1) designed using 4-(4-(4-aminophenyl)piperazin-1-yl)phenol (2) and (S)-N′-(2-(benzyloxy)propylidene)formohydrazide (3) as start materials is successfully obtained via a multistep synthesis and finally characterized by IR, 1H NMR, and single crystal X-ray diffraction. In addition, the in vitro anticancer activities of newly synthesized compound 1 are evaluated against three human bone cancer cell lines U2OS, Saos-2, and GC9811. In addition, the molecular docking is used to study the potential antiviral activity of 1 by calculating the binding sites for the 1AS0 protein.

Bioengineered silver nanoparticles capped with bovine serum albumin and its anticancer and apoptotic activity against breast, bone and intestinal colon cancer cell lines

The aim of the current study was to biosynthesize the silver nanoparticles (AgNPs) from the bacterial strain of Bacillus cereus (ATCC 14579) extracellularly. When bacterial extract was challenged with 1 mM silver nitrate (AgNO3) the color of the extract changed into brown confirms the formation of nanoparticles. These nanoparticles were capped with bovine serum albumin (BSA). UV- visible spectroscopy showed the absorption peak at 420 nm indicates the formation of AgNPs. Fourier Infra –red (FTIR) attenuated total reflection (ATR) spectroscopy showed amide and amine group associated with AgNPs that stabilizes the nanoparticles. Energy dispersive x-ray spectroscopy (EDX) showed a strong peak of silver confirms the presence of silver. Thermo gravimetric analysis (TGA) analysis was used to determine the protein degradation showed less protein degradation at higher temperature confirms the stability of nanoparticles. Transmission electron microscopy (TEM) showed the AgNPs are well dispersed and spherical, and 5.37 nm to 17.19 whereas albumin coated nanoparticles are size ranges from 11.26 nm to 23.85 nm. The anticancer effect of capped AgNPs (cAgNPs) showed the IC50 value against breast cancer MCF-7 at 80 μg/mL, intestinal colon cancer HCT- 116 60 μg/mL, and bone cancer osteosarcoma MG-63 cell line80 μg/mL while against normal fibroblast cells 3T3 cells showed the IC50 value at 140 μg/mL. Lactate dehydrogenase assay (LDH) showed higher toxicity on MCF-7, HCT-116, and MG-63 cells. The apoptotic study clearly showed the blebbing of membrane, chromatin condensation due to the production of reactive oxygen species (ROS) by ethidium bromide and acridine orange dual staining method. The DNA analysis showed the complete fragmentation of the DNA of treated cells when compared with control cells.

Synthesis, crystal structure, anti-bone cancer activity and molecular docking investigations of the heterocyclic compound 1-((2S,3S)-2-(benzyloxy)pentan-3-yl)-4-(4-(4-(4-hydroxyphenyl)piperazin-1-yl) phenyl)-1H-1,2,4-triazol-5(4H)-one

Heterocyclic compound 1-((2S,3S)-2-(benzyloxy)pentan-3-yl)-4-(4-(4-(4-hydroxyphenyl)pipe­razin-1-yl)phenyl)-1H-1,2,4-triazol-5(4H)-one (1) designed using 4-(4-(4-aminophenyl)pipera­zin-1-yl)phenol (2) and (S)-N'-(2-(benzyloxy)propylidene)formohydrazide (3) as start materials is successfully obtained via a multistep synthesis and finally characterized by IR, 1H NMR, and single crystal X-ray diffraction. In addition, the in vitro anticancer activities of newly synthesized compound 1 are evaluated against three human bone cancer cell lines U2OS, Saos-2, and GC9811. In addition, the molecular docking is used to study the potential antiviral activity of 1 by calculating the binding sites for the 1AS0 protein.

Metvan, bis(4,7-Dimethyl-1,10-phenanthroline)sulfatooxidovanadium(IV): DFT and Spectroscopic Study-Antitumor Action on Human Bone and Colorectal Cancer Cell Lines.

The complex bis(4,7-dimethyl-1,10-phenantroline)sulfatooxidovanadium(IV), commonly known as Metvan, was prepared using a known synthetic procedure. Its optimized molecular structure was obtained by DFT calculations, as it was impossible to grow single crystals adequate for a crystallographic study. The complex was also characterized by a detailed analysis of its infrared spectrum, supported by the theoretical calculations, and also by some data derived from its Raman spectrum. In addition, cytotoxicity studies were performed using human osteosarcoma (MG-63) and human colorectal adenocarcinoma (HT-29) cell lines. The results show that Metvan impaired cell viability of both cancer cell lines in a low concentration range (0.25-5.0μM).

Genetic Alphabet Expansion Provides Versatile Specificities and Activities of Unnatural-Base DNA Aptamers Targeting Cancer Cells

The potential of genetic alphabet expansion technologies using artificial extra base pairs (unnatural base pairs) has been rapidly expanding and increasing. We present that the hydrophobic unnatural base, 7-(2-thienyl)imidazo[4,5-b]pyridine (Ds), which acts as a fifth letter in a DNA library, provides a series of high-affinity DNA aptamers with versatile binding specificities and activities to cancer cells. These Ds-containing DNA aptamers were generated by a method called cell-ExSELEX to target three breast cancer cell lines: MCF7, MDA-MB-231, and T-47D. Aptamer 14A-MCF7, which targets MCF7 cells, specifically binds to MCF7 cells, but not other cancer cell lines. Aptamer 07-MB231, which targets MDA-MB-231 cells, binds to a series of metastatic bone and lung cancer cell lines. Aptamer 05-MB231 targets MDA-MB-231 cells, but it also binds to all of the cancer and leukemia cell lines that we examined. None of these aptamers bind to normal cell lines, such as MCF10A and HUVEC. In addition, aptamers 14A-MCF7 and 05-MB231 are internalized within the cancer cells, and aptamer 05-MB231 possesses anti-proliferative properties against most cancer cell lines that we examined. These aptamers and the generation method are broadly applicable to cancer cell imaging, biomarker discovery, cancer cell profiling, anti-cancer therapies, and drug delivery systems.

Design and Development of Polysaccharide-Doxorubicin-Peptide Bioconjugates for Dual Synergistic Effects of Integrin-Targeted and Cell-Penetrating Peptides for Cancer Chemotherapy.

Polymer-drug conjugation is an attractive approach for target delivering insoluble and highly toxic drugs to tumor sites to overcome the side-effects caused by cancer chemotherapy. In this study we designed and synthesized novel polymer-drug-peptide conjugates for improved specificity on targeting cancer cells. Chemically modified polysaccharide, carboxymethylcellulose (CMC), was conjugated with doxorubicin (DOX) anticancer drug by amide bonds and dually biofunctionalized with integrin-target receptor tripeptide (RGD) and l-arginine (R) as cell-penetrating amino acid for synergistic targeting and enhancing internalization by cancer cells. These bioconjugates were tested as prodrugs against bone, breast, and brain cancer cell lines (SAOS, MCF7, and U87) and a normal cell line (HEK 293T, reference). The physicochemical characterization showed the formation of amide bonds between carboxylates (-RCOO-) from CMC biopolymer and amino groups (-NH2) from DOX and peptides (RGD or R). Moreover, these polymer-drug-peptide bioconjugates formed nanoparticulate colloidal structures and behaved as "smart" drug delivery systems (DDS) promoting remarkable reduction of the cytotoxicity toward normal cells (HEK 293T) while retaining high killing activity against cancer cells. Based on cell viability bioassays, DNA-staining, and confocal laser microscopy, this effect was assigned to the association of physicochemical aspects with the difference of the endocytic pathways and the drug release rates in live cells caused by the biofunctionalization of the macromolecule-drug systems with RGD and l-arginine. In addition, chick chorioallantoic membrane (CAM) assay was performed as an in vivo xenograft model test, which endorsed the in vitro results of anticancer activities of these polymer-drug systems. Thus, prodrug nanocarriers based on CMC-DOX-peptide bioconjugates were developed for simultaneously integrin-targeting and high killing efficacy against cancer cells, while preserving healthy cells with promising perspectives in cancer chemotherapy.

Anti-metastatic Potentiality of Purified Anthocyanin from Osbeckia aspera (L.) Blume. and O. reticulata Bedd. Against Selected Human Cancer Cell Lines

To evaluate the anti-metastatic potentialities of purified anthocyanin from Osbeckia aspera (L.) Blume. and O. reticulata Bedd. against selected human cancer cell lines such as HT29 colon, MG63 bone and HeLa cervical by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, apoptosis and DNA fragmentation test. Anthocyanin was extracted from the in vitro callus culture of the Osbeckia species, purified using amberlite column chromatography and fractionated by LC-MS/MS. Anthocyanin producing callus cultures were trialed on MS medium fortified with various combinations of phytohormones and sucrose. Significant callus formation in O. aspera was initiated in cultures containing 0.5 mg/L of 2, 4-D and 0.5 mg/L BA, while that in O. reticulata was initiated with 1.2 mg/L BA and 1.4 mg/L NAA. The same hormonal combinations on sub-culturing turned white friable callus into red compact callus. Purified anthocyanins obtained from O. aspera and O. reticulata contained Malvidin-3 -diglucoside, delphinidin, cyanindin aglycone and Peonidin. Osbeckia species displayed differential responses against the HT29 colon, MG63 bone and HeLa cervical cancer cell lines in terms of IC50 values of toxicity. O. aspera was more effective against HeLa cervical cell lines (23.7μg/ml) followed by HT29 colon (64.7μg/ml) as compared to O. reticulata. Poor selectivity index was noticed with bone cancer cell lines. The results were substantiated by apoptotic analysis and DNA fragmentation results. The overall results suggest that the purified anthocyanin of O. aspera and O. reticulata was excellent as antimetastatic and warrant further studies to isolate novel compounds for chemotherapeutic use.

Bmi‑1‑targeting suppresses osteosarcoma aggressiveness through the NF‑κB signaling pathway.

Bone cancer is one of the most lethal malignancies and the specific causes of tumor initiation are not well understood. B-cell-specific Moloney murine leukemia virus integration site 1 protein (Bmi-1) has been reported to be associated with the initiation and progression of osteosarcoma, and as a prognostic indicator in the clinic. In the current study, a full-length antibody targeting Bmi-1 (AbBmi-1) was produced and the preclinical value of Bmi-1-targeted therapy was evaluated in bone carcinoma cells and tumor xenograft mice. The results indicated that the Bmi-1 expression level was markedly upregulated in bone cancer cell lines, and inhibition of Bmi-1 by AbBmi-1 reduced the invasiveness and migration of osteosarcoma cells. Overexpression of Bmi-1 promoted proliferation and angiogenesis, and increased apoptosis resistance induced by cisplatin via the nuclear factor-κB (NF-κB) signal pathway. In addition, AbBmi-1 treatment inhibited the tumorigenicity of osteosarcoma cells in vivo. Furthermore, AbBmi-1 blocked NF-κB signaling and reduced MMP-9 expression. Furthermore, Bmi-1 promoted osteosarcoma tumor growth, whereas AbBmi-1 significantly inhibited osteosarcoma tumor growth in vitro and in vivo. Notably, AbBmi-1 decreased the percentages of Ki67-positive cells and terminal deoxynucleotidyl transferase dUTP nick end labeling-positive cells in tumors compared with Bmi-1-treated and PBS controls. Notably, MMP-9 and NF-κB expression were downregulated by treatment with AbBmi-1 in MG-63 osteosarcoma cells. In conclusion, the data provides evidence that AbBmi-1 inhibited the progression of osteosarcoma, suggesting that AbBmi-1 may be a novel anti-cancer agent through the inhibition of Bmi-1 via activating the NF-κB pathway in osteosarcoma.

Abstract 4147A: Potent anti-cancer activity of the imipridone ONC206: A selective dopamine D2-like receptor antagonist

Abstract DRD2 is a G protein-coupled receptor (GPCR) that is overexpressed in many cancers, controls an array of pro-survival signaling pathways, and its antagonism causes anti-cancer effects. ONC201, the founding member of the imipridone class of anti-cancer compounds, is a small molecule DRD2 antagonist that is in Phase I/II advanced cancer clinical trials. In this study, we evaluated the binding target and antitumor activity of ONC206, a chemical analogue of ONC201. An orphan small molecule target prediction algorithm revealed that ONC206, like ONC201, antagonizes DRD2. Experimental GPCR profiling using the PathHunter® β-Arrestin assay, determined that ONC206 selectively antagonizes the D2-like (DRD2/3/4), but not the D1-like (DRD 1/5), subfamily of dopamine receptors. ONC206 possesses a ~10-fold increased affinity for DRD2 compared to ONC201 with a Ki of ~320nM with selectivity that was superior to approved antipsychotics. The increased association rate for the ONC206-DRD2 interaction was responsible for the increased affinity, whereas the dissociation rate was similar to ONC201 and atypical antipsychotics that are well tolerated. TCGA analysis and immunohistochemistry of patient-derived tissue microarrays revealed DRD2 was overexpressed in neuroblastoma, sarcoma and pheochromocytoma specimens relative to normal tissues. In vitro efficacy profiling of ONC206 in the Genomic of Drug Sensitivity in Cancer collection of cell lines revealed broad efficacy across most tumor types (GI50 <78-889nM). Bone cancer and neuroblastoma were identified as the most ONC206-responsive solid tumor types that were comparatively less responsive to ONC201. Within bone cancer cell lines, Ewing’s sarcoma (n=16) was the most sensitive to ONC206 with nanomolar sensitivity (GI50 168-303nM) that was superior to ONC201. ONC206 was highly efficacious in neuroblastoma (n=35, GI50 87-589nM) including cell lines derived from metastatic sites and with MYCN amplification associated with poor prognosis. In the PC12 rat pheochromocytoma cell line ONC206 (GI50 200nM) was superior to ONC201. ONC206 time-course experiments revealed anti-cancer effects occurring at 48-72 post-treatment, similar to ONC201. In support of a wide therapeutic window, ONC206 reduced the viability of normal fibroblasts (HFF-1) at relatively high doses (GI50 > 5µM). Efficacy evaluations in MHH-ES-1 athymic nude mice xenografts demonstrated that ONC206 (100 mg/kg PO every 10 days) causes significant tumor growth inhibition that was comparable to methotrexate (400 mg/kg, IP) while being better tolerated. In summary, ONC206 is an imipridone that acts as a selective antagonist of DRD2 at nanomolar concentrations and has broad-spectrum anti-tumor activity. ONC206 may address tumor types where the properties of ONC201 do not permit for complete therapeutic engagement in vivo. Citation Format: Varun Vijay Prabhu, Neel Madhukar, Jessica Wagner, Rohinton Tarapore, Mathew Garnett, Ultan McDermott, Cyril Benes, Neil Charter, Sean Deacon, Alexander VanEngelenburg, Olivier Elemento, Wafik El-Deiry, Martin Stogniew, Wolfgang Oster, Joshua Allen. Potent anti-cancer activity of the imipridone ONC206: A selective dopamine D2-like receptor antagonist [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4147A. doi:10.1158/1538-7445.AM2017-4147A

Anti-proliferative and anti-metastasis effects of ten oligostilbenes from the seeds of Paeonia suffruticosa on human cancer cells.

Paeonia suffruticosa from the section Moutan of the genus Paeonia is an important Chinese medicinal herb. In our previous study, 10 oligostilbenes from the seeds of P. suffruticosa, including trans-resveratrol and its dimers and trimers, were isolated and identified. In the present study, the anti-proliferative effects of these 10 oligostilbenes were systemically evaluated in a panel of human lung, breast and bone cancer cell lines, and their apoptotic effects were analyzed using a high-content multiplex apoptosis assay and a fluorescent caspase-3/7 assay. Furthermore, their anti-metastasis effects were examined in an invasive breast cancer cell line. Among the ten compounds, two resveratrol dimers, trans- and cis-gnetin H, showed the most potent anti-proliferative and anti-metastasis effects. All trans-oligostilbenes were more effective than their cis-forms, and trimers of resveratrol were more effective than dimers and the resveratrol isomer. The structure-activity relationships revealed that the polymerization degree, the double bond in the stilbene skeleton, and the steric arrangement and conformation of oligostilbenes obviously affected their antitumor potential. The results from this study provide valuable information for future semi-synthesis of resveratrol derivatives to develop novel cancer chemopreventive agents.

Temporary Single-Cell Coating for Bioprocessing with a Cationic Polymer.

Temporary single-cell coating is a useful tool for cell processing, allowing manipulation of cells to prevent cell attachment and agglomeration, before re-establishing normal cell function. In this work, a speckled coating method using a known polycation [poly(l-lysine), PLL] is described to induce cell surface electrostatic charges on three different cell types, namely, two bone cancer cell lines and fibroblasts. The morphology of the PLL speckled coating on the cell surface, internalization and metabolization of the polymer, and prevention of cellular aggregations are reported. Polymer concentration was found to be the key parameter controlling both capsule morphology and cell health. This approach allows a temporary cell coating over the course of 1–2 h, with cells exhibiting phenotypically normal behavior after ingesting and metabolizing the polymer. The process offers a fast and efficient alternative to aid single-cell manipulation for bioprocessing applications. Preliminary work on the application of PLL speckled cell coating in enabling reliable bioprinting is also presented.

Keratin 13 expression reprograms bone and brain metastases of human prostate cancer cells.

Lethal progression of prostate cancer metastasis can be improved by developing animal models that recapitulate the clinical conditions. We report here that cytokeratin 13 (KRT13), an intermediate filament protein, plays a directive role in prostate cancer bone, brain, and soft tissue metastases. KRT13 expression was elevated in bone, brain, and soft tissue metastatic prostate cancer cell lines and in primary and metastatic clinical prostate, lung, and breast cancer specimens. When KRT13 expression was determined at a single cell level in primary tumor tissues of 44 prostate cancer cases, KRT13 level predicted bone metastasis and the overall survival of prostate cancer patients. Genetically enforced KRT13 expression in human prostate cancer cell lines drove metastases toward mouse bone, brain and soft tissues through a RANKL-independent mechanism, as KRT13 altered the expression of genes associated with EMT, stemness, neuroendocrine/neuromimicry, osteomimicry, development, and extracellular matrices, but not receptor activator NF-κB ligand (RANKL) signaling networks in prostate cancer cells. Our results suggest new inhibitors targeting RANKL-independent pathways should be developed for the treatment of prostate cancer bone and soft tissue metastases.

Flower-shaped gold nanoparticles synthesized using Kedrostis foetidissima and their antiproliferative activity against bone cancer cell lines

Three different methods were employed for the synthesis of biogenic gold nanoparticles using the aqueous extracts of Kedrostis foetidissima. The interaction of gold nanoparticles with the phytoconstituents was investigated by FTIR. The complete reduction of chloroaurate ions to gold nanoparticles was monitored using UV–visible spectroscopy under the different plant extract concentration and conditions. The formation of gold nanoparticles was confirmed using XRD, SEM and TEM analysis. The anisotropic and flower-shaped gold nanoparticles of size below 25 nm was confirmed by TEM analysis. Cucurbitacins, the chief constituents of K. foetidissima probably might have interacted with the chloroaurate ions, facilitating the formation of gold nanoparticles. KFL-mediated AuNPs showed 88 % cell viability against bone cancer cell lines at 200 µg/ml concentration using MTT assay. This may be due to the formation of flower-shaped nature of KFL-mediated AuNPs. The novelty of the work lies in the plant-mediated synthesis of biocompatible gold nanoparticles of size less than 25 nm showing 88 % of cell viability against bone cancer cell lines.

Antitumor evaluation of two selected Pakistani plant extracts on human bone and breast cancer cell lines.

BackgroundThe medicinal plants Vincetoxicum arnottianum (VSM), Berberis orthobotrys (BORM), Onosma hispida (OHRM and OHAM) and Caccinia macranthera (CMM) are used traditionally in Pakistan and around the world for the treatment of various diseases including cancer, dermal infections, uterine tumor, wounds etc. The present study focuses on the investigation of the selected Pakistani plants for their potential as anticancer agents on human bone and breast cancer cell lines in comparison with non-tumorigenic control cells.MethodsThe antitumor evaluation was carried out on human bone (MG-63, Saos-2) and breast cancer cell lines (MCF-7, BT-20) in contrast to non-tumorigenic control cells (POB, MCF-12A) via cell viability measurements, cell cycle analysis, Annexin V/PI staining, microscopy based methods as well as migration/invasion determination, metabolic live cell monitoring and western blotting.ResultsAfter the first initial screening of the plant extracts, two extracts (BORM, VSM) revealed the highest potential with regard to its antitumor activity. Both extracts caused a significant reduction of cell viability in the breast and bone cancer cells in a concentration dependent manner. The effect of VSM is achieved primarily by inducing a G2/M arrest in the cell cycle and the stabilization of the actin stress fibers leading to reduced cell motility. By contrast BORM’s cytotoxic properties were caused through the lysosomal-mediated cell death pathway indicated by an upregulation of Bcl-2 expression.ConclusionsThe antitumor evaluation of certain medicinal plants presented in this study identified the methanolic root extract of Berberis orthobotrys and the methanolic extract of Vincetoxicum arnottianum as promising sources for exhibiting the antitumor activity. Therefore, the indigenous use of the herbal remedies for the treatment of cancer and cancer-related diseases has a scientific basis. Moreover, the present study provides a base for phytochemical investigation of the plant extracts.Electronic supplementary materialThe online version of this article (doi:10.1186/s12906-016-1215-9) contains supplementary material, which is available to authorized users.