Human Bone Cancer Cells Research Articles

Selective Effects of Cold Atmospheric Plasma on Bone Sarcoma Cells and Human Osteoblasts.

The use of cold atmospheric plasma (CAP) in oncology has been intensively investigated over the past 15 years as it inhibits the growth of many tumor cells. It is known that reactive oxidative species (ROS) produced in CAP are responsible for this effect. However, to translate the use of CAP into medical practice, it is essential to know how CAP treatment affects non-malignant cells. Thus, the current in vitro study deals with the effect of CAP on human bone cancer cells and human osteoblasts. Here, identical CAP treatment regimens were applied to the malignant and non-malignant bone cells and their impact was compared. Two different human bone cancer cell types, U2-OS (osteosarcoma) and A673 (Ewing's sarcoma), and non-malignant primary osteoblasts (HOB) were used. The CAP treatment was performed with the clinically approved kINPen MED. After CAP treatment, growth kinetics and a viability assay were performed. For detecting apoptosis, a caspase-3/7 assay and a TUNEL assay were used. Accumulated ROS was measured in cell culture medium and intracellular. To investigate the influence of CAP on cell motility, a scratch assay was carried out. The CAP treatment showed strong inhibition of cell growth and viability in bone cancer cells. Apoptotic processes were enhanced in the malignant cells. Osteoblasts showed a higher potential for ROS resistance in comparison to malignant cells. There was no difference in cell motility between benign and malignant cells following CAP treatment. Osteoblasts show better tolerance to CAP treatment, indicated by less affected viability compared to CAP-treated bone cancer cells. This points toward the selective effect of CAP on sarcoma cells and represents a further step toward the clinical application of CAP.

Unexpected synthesis, characterization, biological evaluations, and computational details of novel nucleosides containing triazine-pyrrole hybrid

A series of novel nucleosides containing 1,2,4-triazine and pyrrole moieties were efficiently prepared and characterized via analytical, spectral, and computational methods. The synthesized compounds had no effect on human immunodeficiency virus type 1 (HIV-1) (HTLV IIIB). Moreover, the triazine derivatives 8a and 8b were screened for their anticancer activity against five human cancer types [breast adenocarcinoma (MCF-7), pancreatic carcinoma (Paca2), lung carcinoma (A-549), human bone cancer cell line (Hos), and a normal human (BJ-1)] cell lines using MTT assay. The obtained results revealed that the tested nucleosides 8a and 8b had significant inhibitory activity against Paca2, but no activity against A-549, Hos, and BJ-1 cell lines. The structure-activity relationship displayed an increase in anticancer potential of compound 8a with the increase of number of hydrogen bond interactions with Lys 89 as the key interactive amino acid of CDK-2 kinase.

Evaluation of Prebiotic Properties of Galactooligosaccharides Produced by Transgalactosylation Using Partially Purified β-Galactosidase from Enterobacter aerogenes KCTC2190.

Transgalactosylation reaction is the penultimate step in the production of galactooligosaccharides (GOSs) which has prominent applications in the treatment of disorders. In the present study, partially purified β-galactosidase from Enterobacter aerogenes KCTC2190 was used for the synthesis of prebiotic GOSs. GOSs were produced using lactose as substrate. Structural elucidation of collected fractions of GOSs by liquid chromatography electrospray ionization mass spectrometry exhibited the appearance of major peaks of produced GOSs at m/z 241.20, 481.39, 365.11, 527.17, and 701.51 respectively. GOSs facilitated the growth of potential probiotic strains (Lactobacillus delbrueckii ssp. helveticus, Bifidobacterium bifidum, and Lactiplantibacillus plantarum) and liberated propionate and butyrate as principal short-chain fatty acids which established its prebiotic potency. Synbiotic combinations exhibited good antioxidant activities. Synbiotic combinations also exhibited antimicrobial activities against pathogenic microorganisms namely Staphylococcus aureus and Escherichia coli. Synbiotic combinations of GOSs and the respective probiotic microorganisms were able to decrease viable human bone cancer cells (MG-63).

In silico and in vitro analysis of FAK/MMP signaling axis inhibition by VO-clioquinol in 2D and 3D human osteosarcoma cancer cells.

The study of novel mechanisms of action of vanadium compounds is critical to elucidating the role and importance of these kinds of compounds as antitumor and antimetastatic agents. This work deals with in silico and in vitro studies of one clioquinol oxidovanadium(iv) complex [VO(clioquinol)2], VO(CQ)2, and its regulation of FAK. In particular, we focus on elucidating the relationship of the FAK inhibition, MMP activity and antimetastatic effects of the complex in human bone cancer cells.

JAK1/STAT3 regulatory effect of β-caryophyllene on MG-63 osteosarcoma cells via ROS-induced apoptotic mitochondrial pathway by DNA fragmentation.

Currently, available treatment for osteosarcoma is combinational chemotherapy of doxorubicin, cisplatin, and methotrexate before and after surgery with overall 5-year survival rate of less than 40%. The present study was aimed to assess the anticancer effects of a phytochemical named β-caryophyllene (BCP) in treating osteosarcoma. We assessed the effect of (BCP) on oxidative stress, proliferation, apoptosis, and inflammation in human bone cancer cells MG-63. Our results showed that BCP induced reactive oxygen species (ROS) generation at 20 µM concentration in MG-63 cells. The same dose was also shown to exhibit proapoptotic and antiproliferative effects in bone cancer cells MG-63. We demonstrated that the treatment of MG-63 cells with BCP prompted mitochondrial apoptosis via upregulation of Bax and caspase-3 and downregulation of Bcl-2 as well as prompted mitochondrial membrane potential. Our results also showed stimulation of Janus kinase 1/signal transducer and activator of transcription 3 (JAK1/STAT3) signaling pathway in bone cancer MG-63 cells upon BCP treatment along with the induction of proinflammatory genes at the messenger RNA level. Overall results suggest that the treatment of MG-63 cells with BCP promotes apoptosis and inflammation via ROS and JAK1/STAT3 signaling pathway.

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.

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.

RETRACTED: A new heterocyclic compound initiates ROS accumulation and cause apoptotic cell death in human bone cancer cells

Abstract A new heterocyclic compound 7-methoxy-6-(3-morpholinopropoxy)quinazolin-4(3H)-one (1), designed using methyl 5-hydroxy-4-methoxy-2-nitrobenzoate (2) as the starting material, was successfully obtained via multiple synthesis route and finally characterized by fourier transform infrared (FT-IR) spectroscopy, 1H nuclear magnetic resonance (NMR), and single crystal X-ray crystallography. We investigated its effect on cell viability and proliferation with Cell Counting Kit-8 (CCK8) assay. The results revealed that compound 1 could block the proliferation of Saos-2 bone cancer cells. In addition, Annexin V-FITC/PI assay and Western blot analysis were performed to detect whether compound 1 could induce cell apoptosis. The results indicated that compound 1 could increase the number of apoptotic cells remarkably. Moreover, we examined the impact of compound 1 on ROS generation. Results revealed that compound 1 up-regulated the reactive oxygen species (ROS) genes expression and promoted the accumulation of ROS in Saos-2 cells. Finally, molecular docking has been utilized to study the binding mode of compound 1 with tubulin.

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.

Synthesis, characterization and in vitro anti-cancer activity of vanadium-doped nanocrystalline hydroxyapatite

Nanocrystalline V(v)-doped hydroxyapatite and its reduced analogue (V(v) and V(iv) mixture) show promising in vitro cytotoxicity against cultured human bone cancer cells.

Fabrication and characterization of nanobiocomposite scaffold of zein/chitosan/nanohydroxyapatite prepared by freeze-drying method for bone tissue engineering

In this investigation, porous composite scaffolds were prepared using a freeze-drying procedure by mixing zein (ZN), chitosan (CS) and nanohydroxyapatite (nHAp) in different inorganic/organic weight ratios. The gained nanocomposite scaffolds were studied using X-ray diffractometer (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), and N2-adsorption-desorption technique. Also, swelling, porosity, mechanical properties, biomineralization capability, degradation, cell attachment, and cell viability of the composite scaffolds were studied. The results showed a porous nature with acceptable pore dimensions and interconnections for cell penetration and colonization. In addition, the cytocompatibility of the ZN/CS/nHAp scaffolds was surveyed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) evaluation and cell attachment studies using human bone cancer cells. Studies indicated non- toxicity to the cells, and the cells were found to be attached to the pore walls within the scaffolds. The results related to physicochemical properties and superior cytocompatibility suggested that ZN/CS/nHAp scaffold could be potential candidate materials for the tissue engineering.

EVALUATION OF ANTIMICROBIAL, CYTOTOXICITY, AND DYEING PROPERTIES OF PRODIGIOSIN PRODUCED BY SERRATIA MARCESCENS STRAIN JAR8

Objective: Prodigiosin is a red tripyrrole pigment which is synthesized as a secondary metabolite by biotypes A1, A2, and A6 of Serretia marcescens. This study was aimed to optimize production parameters for prodigiosin pigment produced by S. marcescens strain JAR8 and to determine its antimicrobial, cytotoxicity, and dyeing properties.Methods: The effect of various media components, process parameters (temperature, pH, incubation period), and other supplements was investigated, and the maximum production of prodigiosin was found to be at temperature 28°C, pH 7.0, incubation period of 5 day with peptone in the media. Antibacterial, cytotoxicity, and dyeing properties of prodigiosin were studied in depth.Results: The antimicrobial study of prodigiosin revealed that it is a potent inhibitor of Gram-positive bacteria as well as Gram-negative bacteria. The prodigiosin exhibited commendable dose-dependent cytotoxicity effect against human bone cancer cells with the IC50 of 108 μg/ml. Prodigiosin produced by strain JAR8 resisted the action of acid, alkali and detergent when applied to textile cloth.Conclusion: The deep red pigment prodigiosin produced by strain JAR8 is economically effective and can be categorized as microbial natural product for further pharmaceutical applications.

Bufalin Induces Apoptosis of Human Osteosarcoma U-2 OS Cells through Endoplasmic Reticulum Stress, Caspase- and Mitochondria-Dependent Signaling Pathways.

Bone cancer is one of the cancer-related diseases, and there are increased numbers of patients with bone cancer worldwide. Therefore the efficacy of treatment of bone cancer is considered extremely vital. Bufalin has been showed to have biological activities including anticancer activities in vitro and in vivo. However, the exact associated mechanisms for bufalin induced apoptosis in human bone cancer cells are still unclear. In the present study, we investigated the effect of bufalin on the cytotoxic effects in U-2 OS human osteosarcoma cells. For examining apoptotic cell deaths, we used flow cytometry assay, Annexin V/PI double staining, and TUNNEL assay. Reactive oxygen species (ROS), Ca2+, mitochondrial membrane potential (ΔΨm), and caspase-8, -9 and -3 activities were measured by flow cytometry assay. Furthermore, western blotting and a confocal laser microscopy examination were used for measuring the alterations of apoptotic associated protein expression and translocation, respectively. The results indicated that bufalin induced cell morphological changes, decreased the viable cell number, induced apoptotic cell death, and increased the apoptotic cell number, and affected apoptotic associated protein expression in U-2 OS cells. Bufalin increased apoptotic proteins such as Bak, and decreased anti-apoptotic proteins such as Bcl-2 and Bcl-x in U-2 OS cells. Furthermore, bufalin increased the protein levels of cytochrome c (Cyto c), AIF (Apoptosis inducing factor) and Endo G (Endonuclease G) in cytoplasm that were also confirmed by confocal microscopy examination. Based on those findings, bufalin induced apoptotic cell death in U-2 OS cells may be via endoplasmic reticulum (ER) stress, caspase-, and mitochondria-dependent pathways; thus, we may suggest that bufalin could be used as an anti-cancer agent for the treatment of osteosarcoma in the future, and further in vivo studies are needed.

Combined Use of Zoledronic Acid Augments Ursolic Acid-Induced Apoptosis in Human Osteosarcoma Cells through Enhanced Oxidative Stress and Autophagy.

Ursolic acid (UA), a naturally occurring pentacyclic triterpene acid found in many medicinal herbs and edible plants, triggers apoptosis in several tumor cell lines but not in human bone cancer cells. Most recently, we have demonstrated that UA exposure reduces the viability of human osteosarcoma MG-63 cells through enhanced oxidative stress and apoptosis. Interestingly, an inhibitor of osteoclast-mediated bone resorption, zoledronic acid (ZOL), also a third-generation nitrogen-containing bisphosphonate, is effective in the treatment of bone metastases in patients with various solid tumors. In this present study, we found that UA combined with ZOL to significantly suppress cell viability, colony formation, and induce apoptosis in two lines of human osteosarcoma cells. The pre-treatment of the antioxidant had reversed the oxidative stress and cell viability inhibition in the combined treatment, indicating that oxidative stress is important in the combined anti-tumor effects. Moreover, we demonstrated that ZOL combined with UA significantly induced autophagy and co-administration of autophagy inhibitor reduces the growth inhibitory effect of combined treatment. Collectively, these data shed light on the pathways involved in the combined effects of ZOL and UA that might serve as a potential therapy against osteosarcoma.

New dendrimer functionalized multi-walled carbon nanotube hybrids for bone tissue engineering

Two types of multi-walled carbon nanotubes (MWCNTs) functionalized with dendrimer/hydroxyapatite (HAp) nanohybrids viz., MWCNTs-poly(propylene imine) generation 2 (G2)-HAp (MWCNTs-PPI(G2)/HAp) and MWCNTs-poly(propylene imine) generation 3 (G3)-HAp (MWCNTs-PPI(G3)/HAp) were prepared. Initially, pristine-MWCNTs were oxidized to form MWCNTs–COOH and to which PPI(G2) and PPI(G3) were functionalized separately to obtain MWCNTs-PPI(G2) & MWCNTs-PPI(G3) matrices. Nanostructured HAp was prepared by wet precipitation method and bonded to MWCNTs-PPI(G2) & MWCNTs-PPI(G3) matrices. The cross-linkages in carbon nanotube based nanohybrids (CNTs-nanohybrids) were observed by Fourier transform infrared spectroscopy. Surface morphology of the CNTs-nanohybrids and the weight percentage of HAp deposited onto the MWCNTs-PPI(G2 & G3) matrices were confirmed by field emission scanning electron microscopy and energy dispersive spectroscopy analyses. The average crystal size of HAp was calculated from the X-ray diffraction patterns using the Debye–Scherrer equation. The in vitro anticancer effect and cell viability of newly fabricated CNTs-nanohybrids were examined against human bone cancer cell line (human osteosarcoma (MG-63)) by means of 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. The results suggest that the prepared two types of MWCNTs-PPI(G2)/HAp & MWCNTs-PPI(G3)/HAp nanohybrids significantly enhanced the anticancer efficacy in cancer cells. The proposed hybrid materials on the basis of MWCNTs possess valuable scaffolds for both curative and preventative approaches in bone tissue engineering.

Abstract 1182: Ocimum gratissimum aqueous extract induces apoptotic signalling in human osteosarcoma cells.

Abstract Osteosarcoma (Osteosarcoma) is a kind of bone cancer. Eighty percent of this tumor will be metastatic to the lungs or liver. Therefore, these patients generally need to use chemotherapy to improve disease and increase survival possibility. Recently, it has been reported that Ocimum gratissimum (OG) aqueous extract has anti-tumor activity. In this study, we detected the effect of OG extract on human bone cancer cell growth and metastasis. The results showed that the OG extract can inhibit cell growth and induce cell apoptosis in human bone cancer cells, but no effect in normal nerve cell. Therefore, we suggest that OG extract s may have anti-cancer ability, and can provide the use in the development of new anti-cancer drugs Figure . OGE effect on the cell growth in two osteosarcoma cells. Cell growth was determined 0∼3 or 0∼5 days after subculture using the MTT assay as described in Materials and Methods. Absorbance values obtained from the cells on day 0 after subculture were taken as 100%. Citation Format: Chien Chung Lin. Ocimum gratissimum aqueous extract induces apoptotic signalling in human osteosarcoma cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1182. doi:10.1158/1538-7445.AM2013-1182 Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.

Anticancer activity of bovine α‐lactalbumin treated with microbial transglutaminase

This study investigated the effects of bovine α‐lactalbumin (α‐La) treated with microbial transglutaminase on human cancer cells, cell cultures and growth rate assays. The anticancer activity for 10 mg/mL of bovine α‐lactalbumin (α‐La) was measured as ∼90% in a human colorectal cancer cell line HCT 116. For the human bone cancer cell line SJSA‐1, α‐La hydrolysis resulted in higher cytotoxicity compared to untreated tumour cells. The formation of polymers of α‐La was suppressed by the addition of ethylenediaminetetraacetic acid, indicating that polymers of α‐La are promoted by metal ions such as Ca2+. The effect of α‐La on the morphology of SJSA‐1 cells was manifested as morphological changes compatible with apoptosis. Bovine milk α‐La with and without microbial transglutaminase is considered a valuable food ingredient and a nutraceutical for human health.

Transforming growth factor-β-induced mitogenesis of human bone cancer cells

We report for the first time the bifunctional effects of transforming growth factor-β on the growth of cloned human osteosarcoma cells (Htb96). Cell growth was assessed by determining the cell number, replication index and [ 3H]-thymidine incorporation following 48 hours incubation of cultured Htb96 cells with the peptide. Exposure of cells to concentrations of TGF-β upto 40 pM caused a mitogenic response, concentrations between 40 and 800 pM failed to stimulate cell growth and higher doses caused an inhibition of cell proliferation. The initial cell density was found to alter the responsiveness of Htb96 cells to TGF-β; stimulation of proliferation was less profound at high and low cell densities. The observed cell density- and growth factor concentration-dependent effects of TGF-β on the growth of tumour cells might suggest the existence of an autocrine regulatory mechanism. Furthermore, by demonstrating a sensitivity to inhibition by indomethacin, we conclude that the proliferative effect of TGF-β is at least, in part, dependent on the de novo synthesis of prostaglandins.