Human Osteosarcoma Cell Line Research Articles

Selective chromogenic and fluorogenic signalling of Hg2+ ions using a benzothiazole-quinolinyl acrylate conjugate and its applications in the environmental water samples and living cells

We have developed a new chromogenic and fluorogenic chemosensor (L) based on benzothiazole conjugated with quinolinyl acrylate for the detection of Hg2+ ions with excellent selectivity and sensitivity. In the presence of Hg2+, the fluorescence of L was quenched from light blue to colorless. The L could work in the wide pH range of 5.0–8.0 without considerable interference from other tested ions. UV–Visible and fluorometric investigations revealed that the limit of detection (LOD) for Hg2+ was 1.0 μM (202 ppb) and 11.8 nM (2.38 ppb), respectively. Further, the applicability of probe L in the determination of Hg2+ in various water samples and biological samples were tested. Human Bone Osteosarcoma cell lines (HOS) were utilized for this study and satisfying results were obtained. These cell line results also further support the ‘on-off’ properties of probe L. Based on the colorimetric and fluorescent results we have designed a Boolean logic gate for futuristic applications.

Surface modification of zirconia ceramics through cold plasma treatment and the graft polymerization of biomolecules

Although zirconia ceramics were highly versatile as dental implants, their long-term presence in the human body may slow down healing and impede cell growth in the past. To enhance the cytocompatibility of zirconia ceramics, surface activation modification was used to immobilize biopolymers such that a biomimetic environment was created. Hexamethyldisilazane thin films were deposited onto the surface of inorganic zirconia through cold plasma treatment under various power and deposition time settings to form an organosilane interface layer. Next, oxygen plasma treatment was performed to activate the free radicals on the surface. Subsequently, ultraviolet light was employed to graft and polymerize acrylic acid for generating carboxyl groups on the surface. This was followed by a condensation reaction with biopolymers (chitosan, chitosan/poly-γ-glutamic acid, and gelatin). Under a 20-min deposition time at 40W and 150 mTorr, the thin films had a maximum graft density of 2.1mg/cm2. MG-63cells (human osteosarcoma cells) were employed to evaluate cell compatibility. Chitosan and chitosan/poly-γ-glutamic acid promoted the compatibility of MG-63cells (a human osteosarcoma cell line) with zirconia ceramics, whereas gelatin reduced this compatibility. The findings confirm that cold plasma treatment and graft polymerization can promote the immobilization of biomolecules and improve the biocompatibility of zirconia ceramics. This approach can be applied to the modification of zirconia ceramic implants.

Delivery of mGluR5 siRNAs by Iron Oxide Nanocages by Alternating Magnetic Fields for Blocking Proliferation of Metastatic Osteosarcoma Cells.

Although osteosarcoma is the most common primary malignant bone tumor, chemotherapeutic drugs and treatment have failed to increase the five-year survival rate over the last three decades. We previously demonstrated that type 5 metabotropic glutamate receptor, mGluR5, is required to proliferate metastatic osteosarcoma cells. In this work, we delivered mGluR5 siRNAs in vitro using superparamagnetic iron oxide nanocages (IO-nanocages) as delivery vehicles and applied alternating magnetic fields (AMFs) to improve mGluR5 siRNAs release. We observed functional outcomes when mGluR5 expression is silenced in human and mouse osteosarcoma cell lines. The results elucidated that the mGluR5 siRNAs were successfully delivered by IO-nanocages and their release was enhanced by AMFs, leading to mGluR5 silencing. Moreover, we observed that the proliferation of both human and mouse osteosarcoma cells decreased significantly when mGluR5 expression was silenced in the cells. This novel magnetic siRNA delivery methodology was capable of silencing mGluR5 expression significantly in osteosarcoma cell lines under the AMFs, and our data suggested that this method can be further used in future clinical applications in cancer therapy.

Single nucleotide mutation changes the capability of CCN3 in osteosarcoma cell invasion and differentiation

This study aimed to identify significant mutations in CCN3 gene in osteosarcoma, and to explore the influence of this gene on cell invasion and differentiation and the underlying mechanism. Sanger sequencing was used to identify CCN3 gene sequence in human osteosarcoma cell lines, peripheral blood mononuclear cells (PBMC), and osteosarcoma tissues. Wild-type and mutant CCN3 (mCCN3) were ectopically expressed by lentivirus in human osteosarcoma cell lines. Tumor cell invasion was measured by trans-well assay. Osteogenic differentiation was induced by osteogenic differentiating medium and evaluated based on alkaline phosphatase activity and collagen type I alpha 1 chain and osteocalcin expression. Western blotting was used to detect protein levels of CCN3 and mCCN3 in cytoplasmic, nuclear and secreted fractions of cells. A G-to-A single nucleotide mutation in the coding region of CCN3 was found in both osteosarcoma cells and tissues. The frequency of this mutation in osteosarcoma tissue was much higher than that in para-carcinoma tissue and PBMC of healthy people. This nucleotide mutation decreased nuclear glycosylated full length protein level of CCN3 and affected osteosarcoma cell invasion and differentiation. A lower nuclear ratio of glycosylated/non-glycosylated isoforms accounted for the different behavior of mCCN3 compared with CCN3. The G-to-A mutation identified in CCN3 resulted in differential glycosylated full-length protein levels and altered the functional role of CCN3 in osteosarcoma cell invasion and differentiation.

In Vitro and Electrochemical Characterization of Laser-Cladded Ti-Nb-Ta Alloy for Biomedical Applications

Titanium (Ti) and its alloys are predominant choices for use as biomaterials in human implants. Research has shown the adverse effects of using commercial Ti alloy Ti-6Al-4V in the human body, and this presents a need for viable alternatives. In this study, Ti alloy Ti-17Nb-6Ta was manufactured by laser cladding—a prominent additive manufacturing (AM) technology. Laser cladded specimens were evaluated for their in vitro and electrochemical behavior. A human osteosarcoma cell line (MG-63 cells) was used for in vitro investigations. Cell proliferation was good in the physiological medium, and cells were alive when in contact with the laser cladded alloy, even after two to three weeks, indicating good cell viability and compatibility with this alloy. Electrochemical characterization was carried out in Ringer’s solution, and noticeably lower corrosion current density and corrosion rate values were observed. The lower amounts of these parameters indicated the passivation behavior due to multi-layer Ti, Nb, and Ta alloy oxide films. These oxide films also enhanced osseointegration. Thus, the Ti-17Nb-6Ta alloy can be an ideal biocompatible alternative to Ti-6Al-4V.

Physicochemical and biological studies of Ni(II), Cu(II) and Zn(II) ternary complexes of sulfaquinoxaline and 2,2’-bipyrimidine

Three monomeric ternary complexes of Ni(II) (1), Cu(II) (2), and Zn(II) (3) with sulfaquinoxaline (SQO), and 2,2’-bipyrimidine (bpm) as ligands have been prepared and their crystal structures have been determined. The complexes are formulated as M(SQO)2bpm·2H2O, completing MN6 coordination spheres. Hirshfeld surface analysis was employed to study the intermolecular interactions in the crystal lattices of the compounds. From these studies, it was found that hydrogen bonding and π-stacking interactions play an important role. Besides, the complexes have been characterized by FTIR and UV-Visible spectroscopies, and thermogravimetric techniques. The cytotoxicity of the complexes was screened against the tumoral A549 (human lung adenocarcinoma) and MG-63 (human osteosarcoma) cell lines. The complexes showed enhanced antibacterial activity against S. aureus and S. epidermidis as compared with the free SQO ligand.

Phase 2 study of anti-disialoganglioside antibody, dinutuximab, in combination with GM-CSF in patients with recurrent osteosarcoma: A report from the Children's Oncology Group

PurposeNovel effective therapies are urgently needed in recurrent osteosarcoma. GD2 is expressed in human osteosarcoma tumours and cell lines. This study evaluated the disease control rate (DCR) in patients with recurrent osteosarcoma treated with the anti-GD2 antibody dinutuximab plus cytokine therapy as compared to historical outcomes. MethodsAOST1421 was a single-arm Phase 2 study for patients with recurrent pulmonary osteosarcoma in complete surgical remission. Patients received up to five cycles of dinutuximab (70 mg/m2/cycle) with granulocyte-macrophage colony-stimulating factor (GM-CSF). Two different dinutuximab infusion schedules were studied: 35 mg/m2/day over 20 h (2 days) and 17.5 mg/m2/day over 10 h (4 days). Primary end point was DCR, defined as a proportion of patients event free at 12 months from enrolment. The historical benchmark was 12-month DCR of 20% (95% CI 10–34%). Dinutuximab would be considered effective if ≥ 16/39 patients remained event free. Secondary objectives included toxicity evaluation and pharmacokinetics. ResultsThirty-nine eligible patients were included in the outcome analysis. Dinutuximab did not demonstrate evidence of efficacy as 11/39 patients remained event free for a DCR of 28.2% (95% CI 15–44.9%). One of 136 administered therapy cycles met criteria for unacceptable toxicity when a patient experienced sudden death of unknown cause. Other ≥ Grade 3 toxicities included pain, diarrhoea, hypoxia, and hypotension. Pharmacokinetic parameters were similar in the two schedules. ConclusionsThe combination of dinutuximab with GM-CSF did not significantly improve DCR in recurrent osteosarcoma. Dinutuximab toxicity and pharmacokinetics in adolescent and young adult osteosarcoma patients were similar to younger patients. Other strategies for targeting GD2 in osteosarcoma are being developed.

Identification of Stem Cell Related Gene Expression from the Osteosarcoma Cell Core Side

Osteosarcoma is the most frequent primary malignant bone tumor with higher incidences in children and adolescents. Despite clinical evolutions, patients with osteosacoma have had a poor prognosis. There has been increasing evidence that cancer is a stem cell disease. This study sought to isolate and characterize cancer stem cells from human osteosarcoma with relevant literature reviews. Here we show that the emerging evidence suggests osteosarcoma should be regarded as a differentiation disease such as stem cell disease. Two human osteosarcoma cell lines were cultured in non-adherent culture conditions as sarcospheres. Sarcospheres were observed using histomorphology and alkaline phosphatase (ALP) staining. Expression of the embryonic stem cell marker was analyzed with use of reverse transcriptase-PCR. Sarcospheres could be reproduced consistently throughout multiple passages and produced adherent osteosarcoma cell cultures. Expression of stem cell-associated genes such as those encoding Nanog, octamer-binding transcription factor 3/4, sex determining region Y box 2 , c-Myc and ALP indicated pluripotent stem-like cells. These results support the extension of the cancer stem cell theory to include osteosarcoma. Understanding the cancer stem cell derived from human osteosarcoma could lead to the evolution of diagnosis and treatment for osteosarcoma patients.

Green synthesis of silver nanoparticles using Macrolepiota procera extract and investigation of their HSP27, HSP70, and HSP90 inhibitory potentials in human cancer cells

In this study, silver nanoparticles (AgNPs) were synthesized by a microwave-assisted green chemistry method using a crude extract of Macrolepiota procera mushroom. The synthesized M. procera-AgNPs (MP-AgNPs) were characterized by UV–VIS spectroscopy, fourier-transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), dynamic light scattering (DLS) technique, and scanning transmission electron microscopy (S-TEM). The cytotoxic activity of synthesized MP-AgNPs was investigated in human breast cancer (MCF-7), lung cancer (A549), osteosarcoma (Saos-2), and colon cancer (HT-29) cell lines. HSP27, HSP70, and HSP90 inhibitory potential of MP-AgNPs were determined in cancer cells by RT-PCR and Western blotting experiments. To analyze the apoptotic profile of MP-AgNPs in cancer cell lines, the expression levels of apoptotic and anti-apoptotic genes were determined using RT-PCR. The results showed that MP-AgNPs were exhibited anticancer activity in MCF-7, A549, Saos-2, and HT-29 cell lines. MP-AgNPs decreased gene and protein expression levels of HSP27, HSP70, and HSP90 in cancer cells. Also, MP-AgNPs induced apoptotic cell death mechanisms in MCF-7, A549, Saos-2, and HT-29 cell lines. MP-AgNPs are found to be significant metallic nanoparticle systems as efficient HSP inhibitors in the treatment of cancer.

HIF-1α-mediated augmentation of miRNA-18b-5p facilitates proliferation and metastasis in osteosarcoma through attenuation PHF2

Extensive evidence has explored the involvement of microRNAs (miRNAs) in osteosarcoma (OS). Limitedly, the concrete function of microRNA-18b-5p (miR-18b-5p) in OS remains unexplored and largely elusive. Here, we validated that miR-18b-5p significantly elevated in OS via analyzing the data from GEO database. The results showed that miR-18b-5p was overexpressed in human OS tissues and cell lines. The clinical evidence suggested that high level of miR-18b-5p was negatively correlated with the poor prognosis of OS. Meanwhile, miR-18b-5p upregulation facilitated the proliferation and metastasis of OS cells in vitro and in vivo. The mechanism exploration demonstrated that miR-18b-5p acted as a potential inhibitor of PHF2, a tumor suppressor gene, at post-transcriptional level. Moreover, hypoxia induced gene expression of miR-18b-5p was clarified to be transcriptionally mediated by HIF-1α. The clinicopathological analysis in samples of OS patients further supported that miR-18b-5p had a positive correlation with HIF-1α expression, and negative correlation with PHF2. Collectively, the present study uncovered a new molecular mechanism of OS tumorigenesis and development and miR-18b-5p might be a prognostic biomarker and potential therapeutic target for OS treatment.

Controlled and localized drug delivery using Titania nanotubes

Titanium-dioxide nanotubes (TNTs) were fabricated by anodic oxidation of titanium (Ti) substrate and loaded with ciprofloxacin. A thin layer of Poly Lactic-co-Glycolic Acid (PLGA) was coated on top of the drug-loaded TNTs at various thicknesses. Field Emission Scanning Electron Microscopy (FESEM) was used to characterize the developed substrate. The FESEM images of anodized samples showed that, the native oxide layer on the surface of the Ti substrate was replaced with an array of ordered nanotubes up to ~700 nm in length. Drug release studies of ciprofloxacin were conducted in Phosphate Buffered Saline (PBS) solution for 24 h. The studies showed the ability of PLGA to slow down the drug release rate, depending on the thickness of the deposited layer. MG-63 cells (human osteosarcoma cell lines) were cultured on the TNT substrates after loading ciprofloxacin and coating with various concentrations of PLGA to show the potentiality for better osseointegration. An MTT assay was carried out to study cell viability and proliferation on these substrates. Anti-microbial studies were carried out to demonstrate the release of ciprofloxacin in treating infections against Staphylococcus epidermidis. These in-depth studies showed that local concentration of the drug released could be controlled by varying the thickness of the PLGA layer to allow for better implant osseointegration and treatment of bone infections.

Abstract 6308: Antitumor effects of RBPJ Inhibitor-1 on osteosarcoma cells

Abstract Osteosarcoma (OS) is the most common primary malignancy of the bone in adolescents and children with limited treatment options. Drug resistance to current therapy together with severe side effects urgently calls for alternative treatment strategies targeting critical cancer signaling pathways. Recently, our and other studies suggest that hyperactivation of Notch signaling contributes to the pathogenesis of human and murine OS, and its inhibition may be a therapeutic approach for the treatment of OS. Notch pathophysiological activity involves activated Notch (i.e. NICD, the notch intracellular domain) forming a nuclear complex with DNA-binding factors and coactivators such as recombination signal binding protein for immunoglobulin kappa J region (RBPJ). This evolutionary conserved mechanism is involved in the maintenance and survival of cancer stem cells, which can govern various cell activities like metastasis and chemoresistance. This study aims to examine the in vitro effects of targeting the NICD-RBPJ interaction using a small molecular inhibitor of the RBPJ (RIN1) on osteosarcoma cell lines. We found that treatment of 10 µM RIN1 on SJSA-1 (ATCC, CRL-2098), a human osteosarcoma cell line with a high Notch activity, can significantly decrease cell proliferation at 48 and 72 hours (h). We also found that at 10 µM of RIN1 affects SJSA-1 migration using a Boyden chamber-based cell system at 24 h, a time point where we find minimal contribution from an effect on cell proliferation. Treatment with RIN1 can block SJSA-1 clonogenicity, but we showed that it has limited ability to block SJSA-1 cell invasion. Treatment with RIN1 does not initiate or enhance the osteoblast-like differentiation of SJSA-1. Notch target genes HEY1 and CyclinD1 was significantly modulated in SJSA-1 cells upon treatment in a Notch-independent manner. Interestingly, treatment with RIN1 on SJSA-1 decreases SOX2 protein expression, a well-known cancer stem cell marker, and upregulates tumor suppressor RB1 protein expression. Ongoing studies include additional cellular function assays and RNA-seq transcriptomic analysis of RIN1’s effect on two Notch-dependent cancer cell lines including MB 157 (ATCC, CRL-7721) and T29 in its activating context. In summary, RIN1 has been shown to have a significant effect on the different cell behaviors of human osteosarcoma, which may eventually facilitate with the treatment of patients that have metastatic disease. Citation Format: Dakota Callahan, Leetoria Hinojosa, Haydee M. Torres, Jianning Tao. Antitumor effects of RBPJ Inhibitor-1 on osteosarcoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6308.

Abstract 988: Epigenetic upregulation of neuropilin-1 promotes osteosarcoma progression and metastasis

Abstract Background: Osteosarcoma (OS) frequently metastasizes to lung and the prognosis for these patients is grim with 5-year overall survival rates as low as 30%. There has been little change in the treatment paradigm for this disease and new targeted therapies are an urgent and unmet clinical need. Previously, we showed that the epigenetic regulators, histone deacetylase 1 and 2 (HDAC1 and 2) are key mediators of OS progression and lung metastasis and that the HDAC1/2 selective inhibitor (romidepsin) significantly prevented OS lung metastasis in vivo. Yet, inhibition of HDACs by romidepsin has noted toxicities in humans and here we define the potential mechanisms through which HDAC1/2 mediate their effects in a bid to identify additional therapeutic targets. Methods: Human OS cell lines were treated with vehicle or romidepsin for 24h before undergoing RNA/protein isolation for microarray/proteomics to identify differentially regulated genes. Standard siRNA, proliferation, migration and molecular analyses were preformed to investigate the role of candidate genes in OS malignant behavior. Results: Microarray/proteomics analyses on romidepsin treated OS cells identified neuropilin-1 (NRP1), a membrane bound co-receptor for vascular endothelial growth factor, to be highly downregulated in response to romidepsin; (<70% compared to vehicle in multiple OS cell lines). NRP1 knockdown (siRNA) led to reduced cell proliferation and interestingly, increased cell migration. Further, a phosphokinase array revealed increased phosphorylation in eNOS, ERK 1/2 and β-catenin in NRP1 knockdown cells that is consistent with a role for NRP1 in regulating OS migration. Based on this data, we hypothesize that NRP1 is a significant regulator of OS proliferation and migration. Our next steps will be to dissect the role of NRP1 silencing in OS invasion and metastasisin vivo and the potential role of NRP1 in OS differentiation. Conclusions: HDAC1/2 contributes to OS growth potentially via the upregulation of NRP1, a therapeutically actionable target. Citation Format: Niveditha Nerlakanti, Jeremy Mcguire, Diana Yu, Damon Reed, Conor C. Lynch. Epigenetic upregulation of neuropilin-1 promotes osteosarcoma progression and metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 988.

NELL1 Regulates the Matrisome to Promote Osteosarcoma Progression.

NELL1 modulates the sarcoma matrisome to promote tumor growth, invasion, and metastasis, identifying the matrix-associated protein as an orchestrator of cell-ECM interactions in sarcomagenesis and disease progression.

Fabrication and characterization of strontium substituted chitosan modify hydroxyapatite for biomedical applications

The present study aimed to preparation of hydroxyapatite using strontium (Sr2+) and chitosan (CS) for the preparation of porous scaffold material like SrHA-CS. These synthesized nano-materials were alsofurther characterized by Fourier transform infrared spectroscopy and X-ray diffraction analysis to confirm the phase purity and crystallinity. The crystal-like morphology of the SrHA-CS was observed using a high-resolution transmission electron microscope (HRTEM) which revealed rod-like morphology with the major elements confirmed by energy-dispersive X-ray spectroscopy (EDX). The size of the synthesized nanomaterial and their stability were analyzed through dynamic light scattering (DLS)and zeta potential. The hemolytic study indicates that SrHA-CS is non-toxic red blood cells of the humans. Besides, SrHA-CS is also active on disease-causing microorganisms and exhibit a strong zone of inhibition. The cytotoxic (MTT) assay systemically investigatedthe synthesized samplesagainst SaOS-2 human osteosarcoma cell lines it resulting in effectively induce cell proliferation without generating any harmful effect. The acridine orange/Ethidium bromide dual staining (AO/EB) under the fluorescence microscope data suggests that apoptotic cell nuclei became irregularly changed. Our present study findings concluded that the strongly recommend the developed SrHA-CSscaffold excellent materialhas mechanical and functional properties including biological activities while compared to other synthetic nano-materials. Hence, the synthesizedSrHA-CSmaterial can be considered as a promising cost-effective nano-material for human bone regenerative applications.

Circ-RANGAP1/MicroRNA-542-3p/Myosin Regulatory Light Chain Interacting Protein Axis Modulates the Osteosarcoma Cell Progression.

Objective This study is aimed at exploring the influence of circular RNA- (circRNA-) RANGAP1 targeting microRNA- (miR-) 542-3p/myosin regulatory light chain interacting protein (MYLIP) on the biological function of osteosarcoma (OS) cells. Methods Tumor tissues and normal tissues were collected from OS patients and circ-RANGAP1, miR-542-3p, and MYLIP expression was tested by RT-qPCR. The correlation between the clinicopathology/prognosis of patients with OS and circ-RANGAP1 expression was observed. Human OS cell line MG-63 was screened to determine the influences of circ-RANGAP1 and miR-542-3p on OS cell progression. The targeting relation of circ-RANGAP1, miR-542-3p, and MYLIP was probed. Results circ-RANGAP1 expression was elevated in tumor tissues from OS patients, which was correlated to the poor clinicopathology. circ-RANGAP1 expression was augmented in males or patients younger than 20 years old or patients with advanced OS. Higher circ-RANGAP1 expression indicated a poor prognosis in OS patients. After silencing circ-RANGAP1 or elevating miR-542-3p in MG63 cells, cell progression was limited. miR-542-3p downregulation reduced the therapeutic efficacy of silenced circ-RANGAP1. circ-RANGAP1 bound with miR-542-3p to target MYLIP. Conclusion Silenced circ-RANGAP1 boosts MYLIP expression via competitive binding of miR-542-3p to facilitate OS cell progression.

Synthetic approach toward spiro quinoxaline-β-lactam based heterocyclic compounds: Spectral characterization, SAR, pharmacokinetic and biomolecular interaction studies

Series of spiro quinoxaline-β-lactam based heterocyclic compounds (QL 1 – QL 21) were synthesized and characterized by spectroscopic techniques like 1H-NMR, LC-MS, FT-IR spectroscopy and elemental analysis. The binding mode and binding strength between compounds and calf thymus-DNA were estimated by UV–visible spectroscopy, viscosity measurement and molecular docking studies. The compounds bind with the DNA through partial intercalation mode. In the absorption titration experiment, the K b values for all the synthesized compounds were found in the range of 0.24–0.64 × 105 M−1. The protein binding studies of all the synthesized compounds were evaluated by absorption titration experiment, and the K b value for all the compounds was obtained in the range of 0.030–1.571 × 104 M−1. The compounds were screened against two Gram (+ve) and three Gram (–ve) bacteria for antimicrobial activity. The MIC values for all the synthesized compounds were found in 95–255 µM. The LC50 values (cytotoxicity) of the synthesized compounds (QL 1–QL 21) were found in the range of 4.00–12.89 µg/mL. The ADME study was carried out using the online platform SwissADME and admetSAR to evaluate the pharmacokinetic profile of all the synthesized compounds. All the compounds were screened for anticancer activity against the human osteosarcoma (MG-63) cell line. The result shows that all the compounds exhibit effective anticancer activity. Communicated by Ramaswamy H. Sarma

Anti-tumor effects of cryptotanshinone (C19H20O3) in human osteosarcoma cell lines.

Osteosarcoma is the most prevalent malignant bone tumor and occurs most commonly in the adolescent and young adult population. Despite the recent advances in surgeries and chemotherapy, the overall survival in patients with resectable metastases is around 20%. This challenge in osteosarcoma is often attributed to the drastic differences in the tumorigenic profiles and mutations among patients. With diverse mutations and multiple oncogenes, it is necessary to identify the therapies that can attack various mutations and simultaneously have minor side-effects. In this paper, we constructed the osteosarcoma pathway from literature and modeled it using ordinary differential equations. We then simulated this network for every possible gene mutation and their combinations and ranked different drug combinations based on their efficacy to drive a mutated osteosarcoma network towards cell death. Our theoretical results predict that drug combinations with Cryptotanshinone (C19H20O3), a traditional Chinese herb derivative, have the best overall performance. Specifically, Cryptotanshinone in combination with Temsirolimus inhibit the JAK/STAT, MAPK/ERK, and PI3K/Akt/mTOR pathways and induce cell death in tumor cells. We corroborated our theoretical predictions using wet-lab experiments on SaOS2, 143B, G292, and HU03N1 human osteosarcoma cell lines, thereby demonstrating the potency of Cryptotanshinone in fighting osteosarcoma.

Therapeutic Effects of Zoledronic Acid-Loaded Hyaluronic Acid/Polyethylene Glycol/Nano-Hydroxyapatite Nanoparticles on Osteosarcoma.

Zoledronic acid (ZOL) has been approved as the only bisphosphonate for the prevention and treatment of metastatic bone diseases with acceptable safety and tolerability. However, systemic or direct injection of ZOL often causes severe side effects, which limits its clinical application. Here, an innovative nano-drug delivery system, ZOL-loaded hyaluronic acid/polyethylene glycol/nano-hydroxyapatite nanoparticles (HA-PEG-nHA-ZOL NPs), has been found to effectively inhibit the proliferation of three types of human osteosarcoma cell lines (143b, HOS, and MG63) at 1–10 μmol/L, while with low cell cytotoxicity on normal cells. The NPs significantly enhanced the apoptosis-related protein expression and tumor cell apoptosis rate. The NPs could also inhibit the proliferation of osteosarcoma cells by blocking the S phase of the cell cycle. In the orthotopic osteosarcoma nude mice model, local injection of the HA-PEG-nHA-ZOL NPs stimulated tumor necrosis, apoptosis, and granulocyte infiltration in the blood vessels. Altogether, the ZOL nano-delivery system possesses great potential for local treatment to prevent local tumor recurrence and can be applied in clinical osteosarcoma therapy.

PTBP1 as a Promising Predictor of Poor Prognosis by Regulating Cell Proliferation, Immunosuppression, and Drug Sensitivity in SARC.

Background Sarcomas (SARC) have been found as rare and heterogeneous malignancies with poor prognosis. PTBP1, belonging to the hnRNPs family, plays an essential role in some biological functions (e.g., pre-mRNA splicing, cell growth, and nervous system development). However, the role of PTBP1 in SARC remains unclear. In this study, the aim was to investigate the potential role of PTBP1 with a focus on SARC. Methods The expression, prognostic value, possible biological pathways of PTBP1, and its relationship with immune infiltration and drug sensitivity were comprehensively analyzed based on multiple databases. PTBP1 was further validated in osteosarcoma as the most prominent bone SARC. The expression of PTBP1 was investigated through IHC. The prognostic value of PTBP1 was verified in TARGET-OS databases. CRISPR/Cas9-mediated PTBP1 knockout HOS human osteosarcoma cell lines were used to assess the effect of PTBP1 on cell proliferation, migration, metastasis and cell cycle by CCK-8, Transwell migration, invasion, and FACS experiment. Result PTBP1 was highly expressed and significantly correlated with poor prognosis in several cancers, especially in SARC, which was validated in the clinical cohort and osteosarcoma cell lines. The genetic alteration of PTBP1 was found most frequently in SARC. Besides, PTBP1 played a role in oncogenesis and immunity through cell cycle, TGFB, autophagy, and WNT pathways at a pan-cancer level. Knockout of PTBP1 was observed to negatively affect proliferation, migration, metastasis, and cell cycle of osteosarcoma in vitro. Furthermore, PTBP1 was significantly correlated with tumor immune infiltration, DNA methylation, TMB, and MSI in a wide variety of cancers. Moreover, the potential of the expression level of PTBP1 in predicting drug sensitivity was assessed. Conclusions PTBP1 is highly expressed and correlated with prognosis and plays a vital pathogenic role in oncogenesis and immune infiltration of various cancers, especially for SARC, which suggests that it may be a promising prognostic marker and therapeutic target in the future.