Laryngeal Carcinoma Hep-2 Cells Research Articles

Polyvinyl Alcohol Capped Silver Nanostructures for Fortified Apoptotic Potential Against Human Laryngeal Carcinoma Cells Hep-2 Using Extremely-Low Frequency Electromagnetic Field.

: Polyvinyl alcohol-capped silver nanostructures (cAgNSs) were investigated in order to enhance the cytotoxicity, pro-apoptotic, and oxidant patterns of in human laryngeal carcinoma Hep-2 cells by employing a 50 mT electromagnetic field (LEMF) for 30 min. Wet chemical reduction was used to synthesize the cAgNSs, and after they had been capped with polyvinyl alcohol, they were specifically examined for particle size analysis and structural morphology. To visualize how the silver may attach to the protein targets, a molecular docking study was conducted. Estimation of cytotoxicity, cell cycle progression supported by mRNA expression of three apoptotic-promoting genes and one apoptotic-resisting. Particle size analysis results were a mean particle size of 157.3±0.5 nm, zeta potential value of -29.6 mV±1.5 mV, and polydispersity index of 0.31±0.05. Significantly reduction of IC50 against Hep-2 cells by around 6-fold was concluded. Also, we obtained suppression of the proliferation of Hep-2 cells, especially in the G0/G1 and S phases. Significant enhanced mRNA expression revealed enhanced induced CASP3, p53, and Beclin-1 mediated pro-apoptosis and induced NF-κB mediated autophagy in Hep-2 cells. Augmented levels of GR, ROS and MDA as oxidative stress biomarkers were also obtained. HE staining of Hep-2 cells exposed to cAgNSs and LEMF confirmed the enhanced apoptotic potential comparatively. By conclusion, the developed nano-sized structures with the aid of extremely-low frequency electromagnetic field were successful to fortify the anti-cancer profile of cAgNSs in Hep-2 cells.

Citalopram, an antipsychotic agent, induces G1/G0 phase cell cycle arrest and promotes apoptosis in human laryngeal carcinoma HEP-2 cells

Human laryngeal squamous carcinoma (LSCC) is a common malignant tumor in the head and neck. Despite the recently developed therapies for the treatment of LSCC, patients’ overall survival rate still did not enhance remarkably; this highlights the need to formulate alternative strategies to develop novel treatments. The antitumor effects of antidepressant drugs such as citalopram have been reported on several cancer cells; however, they have yet to be investigated against LSCC. The current study was directed to explore the possible antitumor effects of citalopram on human laryngeal carcinoma cell lines (HEP-2). HEP-2 cells were cultured and treated with different doses of citalopram (50–400 µM) for 24, 48, and 72 h. The effects of citalopram on the viability of cancer cells were determined by the MTT assay. In addition, apoptosis and cell cycle analysis were performed by flow cytometry. Moreover, evaluation of the expression of proapoptotic and apoptotic proteins, such as cytochrome c, cleaved caspases 3 and 9, Bcl-2, and BAX, was performed by western blotting analysis. Our results revealed that citalopram significantly suppressed the proliferation of HEP-2 cells through the upregulation of p21 expression, resulting in the subsequent arrest of the cell cycle at the G0/G1 phase. Furthermore, citalopram treatment-induced HEP-2 cell apoptosis; this was indicated by the significant increase of cytochrome c, cleaved caspases 3 and 9, and BAX protein expression. On the contrary, Bcl-2 protein expression was significantly downregulated following treatment with citalopram. The ultrastructure studies were in accordance with the protein expression findings and showed clear signs of apoptosis with ring chromatin condensation upon treatment with citalopram. These findings suggest that citalopram’s anti-tumor activities on HEP-2 cells entailed stimulation of the intrinsic apoptotic pathway, which was mediated via Bcl-2 suppression.

Anticancer effects of punicalagin and 5-fluorouracil on laryngeal squamous cell carcinoma: an <i>in vitro</i> study

The purpose of this study was to assess the apoptotic effects of punicalagin alone and in combination with 5-fluorouracil (5-FU) on laryngeal squamous cell carcinoma (Hep-2) cell line. Hep-2 cells were cultured and divided into four groups: Group 1 received no therapy and served as control, Group 2 received 5-FU only, Group 3 received punicalagin only, and Group 4 received a combination of 5-FU and punicalagin. After 48 hours of incubation, cellular changes were examined under an inverted microscope. The methyl thiazolyl tetrazolium assay, caspase-3 gene level, and vascular endothelial growth factor (VEGF) level were assessed. The control group showed the highest mean value of cancer cell proliferation rate (1.595±0.58), followed by the punicalagin group (1.263±0.447), then the 5-FU group (0.827±0.256), while the combination group showed the lowest proliferation rate (0.253±0.111). The combination group showed the highest mean value of caspase-3 concentration (3.177±0.736), followed by the 5-FU group (1.830±0.646), and punicalagin group (0.741±0.302), while the control group showed the lowest mean value (0.359±0.117). Regarding VEGF levels, the control group had a statistically significant higher mean value, followed by the punicalagin and 5-FU groups, and finally, the combination group which showed the lowest value. Punicalagin exerts an anticancer effect through anti-proliferative action and induction of apoptosis on Hep-2 cell line. Combining punicalagin with 5-FU potentiates its anti-proliferative, apoptotic, and anti-angiogenic actions. It, further, helps in mitigating the putative side effects of 5-FU by reducing the dose required for its therapeutic effects.

Oridonin inhibited epithelial-mesenchymal transition of laryngeal carcinoma by positively regulating LKB1/AMPK signaling.

Oridonin is the main bioactive component of Rabdosia rubescens, and its anticancer activity has been reported in a variety of cancers. However, the molecular mechanism of oridonin in laryngeal carcinoma remains unclear. In the present study, the cytotoxic effect of oridonin on laryngeal carcinoma Hep-2 and TU212 cell lines were initially detected by modified MTT assay. The results showed that oridonin had a dose-dependent anti-proliferative effect on laryngeal carcinoma Hep-2 and TU212 cells. Next, we found that oridonin significantly inhibited the migration and invasion of human laryngeal carcinoma Hep-2 and TU212 cell lines by wound healing assay and transwell assay. Subsequently, the results of quantitative real-time PCR assay and western blotting assay confirmed that oridonin upregulated the expression of E-cadherin while downregulated the expression of N-cadherin in a concentration-dependent manner at mRNA and protein levels. In addition, phosphorylation levels of liver kinase B1 (p-LKB1) and AMP-activated protein kinase (p-AMPK) were also elevated upon oridonin treatment. To further verify the role of LKB1/AMPK signaling pathway in laryngeal carcinoma, overexpression of LKB1 was constructed by plasmid transfection. The data exhibited that overexpression of LKB1 could further reinforce the increase of E-cadherin level and decrease of N-cadherin level mediated by oridonin. Additionally, AMPK inhibitor compound C could reverse anti-metastatic effect of oridonin on laryngeal carcinoma, and antagonise EMT expression. In contrast, AMPK activator AICAR presented the opposite effect. In conclusion, our study revealed that oridonin could remarkably reverse the epithelial-mesenchymal transition of laryngeal carcinoma by positively regulating LKB1/AMPK signaling pathway, which suggested that oridonin may be a potential candidate for the treatment of laryngeal carcinoma in the future.

Magnetic Double Hydrogen Artesunate Nano Liposome on CAL Tongue Squamous Carcinoma and Laryngeal Hep-2-27 Cell Lines: An In Vitro Study

With the continuous development of nanomedicine, scholars have reported many studies on the application of nanocarriers to improve the efficiency of tumor diagnosis and treatment. We designed magnetic nanoparticles combined with liposomes to carry DHA (DHA-MAG-NL) nanoparticles to improve the biocompatibility of DHA, protect DHA activity, and increase DHA blood concentration. To study the inhibitory effect of magnetic dihydroartemisinin nano-liposomes on the proliferation of human tongue squamous cell carcinoma CAL-27 cell line and laryngeal carcinoma Hep-2 cell line In Vitro. The effect of DHA-Mag-NL concentration gradient on the proliferation of Hep-2 and Cal-27 cells at different intervention times was detected by MTT assay. The effects of DHA concentration of 40 μM on the cell cycle and apoptosis of Hep-2 and Cal-27 were analyzed by flow cytometry. Statistical analysis showed that DHA-Mag-NL had a weak inhibitory effect on Cal-27 cells. The apoptosis-promoting effect of DHA-Mag-NL on Cal-27 tumor cells was weaker than that of DHA for 24 h. We concluded that DHA-Mag-NL can inhibit the proliferation of Hep-2 and Cal-27 tumor cells and promote apoptosis, which is dependent on drug dose and intervention time.

Photostability, cytotoxicity, and photothermal impact of AgNPs, CoAgNC, and IOAgNC on HEp-2 laryngeal carcinoma cells

Magnetic nanoparticles have shown promise in various medical applications, including cancer treatment and diagnostics. The objective of this study is to attain mastery over the synthesis of silver nanoparticles (AgNPs), Cobalt silver nanocomposite (CoAgNC) and magnetite silver nanocomposite (IOAgNC), while also examining their photostability, cytotoxicity, and photothermal impact on Human Laryngeal Carcinoma Cell line (HEp-2). The preparation and characterization of AgNPs, CoAgNC, and IOAgNC were described, including the assessment of their photostability and cytotoxicity on HEp-2 cell line. The photostability of these nanomaterials was determined by subjecting them to different light sources, while their morphology and magnetic properties were characterized using a spectrophotometer, Transmission Electron Microscopy (TEM), and Vibrating Sample Magnetometer (VSM). The results of the characterization of AgNPs, CoAgNC, and IOAgNC showed that the prepared nanomaterials have distinct optical and morphological properties. The synthesized nanomaterials were stable in aqueous solutions and exhibited magnetic properties. The cytotoxicity tests on HEp-2 cells showed that the nanomaterials were not toxic in the dark, but their toxicity increased under light exposure, with IOAgNC showing the most significant toxicity. CoAgNC was found to be photostable under UV light and Light Emitting Diode (LED) irradiation due to the little lattice mismatch between cobalt and silver. Nanocomposite systems of high magnetic moment, such as IOAgNC and CoAgNC, have potential for tumor treatment through magnetic fluid hyperthermia. The photostability, chemical stability, and biocompatibility of AgNPs, CoAgNC, and IOAgNC were investigated and found to be non-toxic even at high concentrations. These nanomaterials have multiple optical and magnetic functionalities and are promising candidates for biomedical applications.

Perillaldehyde Building Block’s Derivatives: Targeted Bioactivities and Molecular Docking

Background:Recently, the essential oil (EO) of Ammodaucus leucotrichus has attracted particular attention in hemi-synthesis due to the reactivity of its main monoterpenes.Objective:In vitro cytotoxic and antileishmanial activities of the EO and hemi-synthesized compounds were evaluated, and an in-silico simulation was conducted.Methods:The EO was characterized by GC-MS. Two benzodiazepines (C1, C2), benzimidazole (C3), and Schiff base (C4) were hemi-synthesized by an in situ condensation of its major monoterpenes (perillaldehyde) with different amine derivatives. Synthesized compounds and the EO were evaluated for their cytotoxic activity against laryngeal carcinoma-Hep2 and rhabdomyosarcoma-RD cell lines and for their antileishmanial activity against Leishmania (L) major. Molecular docking was performed to pin the binding interactions of active molecules.Results:A significant cytotoxic effect of benzodiazepines C1 and C2 was noted against Hep2 cells (IC50/C1 = 39.48 ±0.7 μM, IC50/C2 = 48.82±1.33 μM) and RD (IC50/C1 = 15.96±1.99 μM; IC50/C2 = 74.3±2.44 μM), while the EO highly inhibited Hep2 cell lines (IC50/EO = 17.3±0.85 μg/mL). C4 strongly inhibited L. major growth with an IC50 value of 8.00±1.37 Μμ. The docking scores of ligand-C4 on L. major pteridine reductase and L. major methionyl-tRNA synthetase and ligands-(C1, C2) on the human protein kinase and epidermal growth factor kinase presented significant affinity to the receptor active sites (ΔG from -7.7 to -8.8 kcal/mol).Conclusion:The cytotoxic effect of derivatives C1, C2, and EO, as well as the anti-parasitic effect of C4, may suggest them as effective bioactive agents or pharmaceutical probes.

Norsquaraine endowed with anticancer and antibacterial activities

BackgroundPhotodynamic therapy (PDT) is a method for the treatment of cancer. Furthermore, PDT can also be used for the eradication of bacteria. The photo-sensitizing drug, a.k.a photosensitizer, is critical for the success of PDT. Although norsquaraines are analogs of squaraine dyes, they are overlooked as photosensitizers. MethodsIn this work, synthesis, characterization, bioimaging and in vitro PDT applications of a new norsquaraine dye 1 were described. We also prepared nanoparticles from norsquaraine 1 and Pluronic F127 to obtain 1@F127. ResultsNorsquaraine 1 boosted the generation of reactive oxygen species over a wide range of pH (pH 8.0, 7.0, 6.0, and 2.2.). Furthermore, 1 was internalized by epidermoid laryngeal carcinoma Hep-2 (Hep-2) cells and used for fluorescence imaging. Remarkably, norsquaraine 1 destroyed most of the cancer cells (ca. 77% to 89%) after illumination with red light. Most strikingly, 1 successfully inhibited the growth of Methicillin-resistant Staphylococcus aureus (MRSA) upon illumination. Last but not least, photodynamic sterilization of tomato juice, an acidic beverage, was feasible using 1 as a photo sterilizer. Nano formulation of 1 with Pluronic F127 provided 1@F127 nanoparticles. It is lucid that 1@F127 nanoparticles permeate into Hep-2 cells and boost the generation of ROS upon illumination. ConclusionNorsquaraine 1 shows superior features as a photosensitizer pertinent to PDT in a wide range of pH. This norsquaraine is endowed with anticancer and antibacterial activities. Which should be further evaluated.

КЛИНИКО-ДИАГНОСТИЧЕСКОЕ ЗНАЧЕНИЕ АНТИНУКЛЕАРНЫХ АНТИТЕЛ В ПРАКТИКЕ ПЕДИАТРА (ЧАСТЬ I)

In recent decades, there has been an increase in the incidence of various autoimmune, including rheumatic diseases, while their debut has been shifted to an earlier age. The clinical picture of rheumatic diseases is distinguished by severe polymorphism; therefore, laboratory diagnostic methods are important for diagnostic search. One of the most widely used tests in the rheumatological practice is the detection and identification of antinuclear antibodies (ANA). ANA are a large group of autoantibodies (auto-Abs) directed against various cell structures, including not only the nucleus but also the nuclear membrane, mitotic apparatus, cytoplasm components and cell organelles, as well as cell membranes. Different methods are used for testing, but the method of ANA detection through indirect immunofluorescence antinuclear antibody test (IF-ANA) is recommended as a “gold standard” with human laryngeal carcinoma HEp-2 cells being recognized as a standard substrate. IF-ANA with HEp-2 has the highest diagnostic sensitivity compared with other methods, it includes the phase of serial dilutions of positive serums and a visual analysis of the type of glow, the list of which currently has 30 patterns. ANA glow types are caused by various auto-Abs that respond with antigens located in the nucleus and cytoplasm of the HEp-2 cells line. Assessment of the type of glow in conjunction with clinical and laboratory data allows planning of further examination with confirming tests to determine the specificity of ANA, some of which, in particular, are auto-Abs to DFS70 antigen, which in the absence of other types of autoantibodies, are more likely to doubt the presence of a systemic rheumatic disease. The diagnostic specificity of various screening techniques for ANA detection is relatively low, they are detected with a fairly high frequency in patients with other diseases and in healthy individuals of different gender and age, including children.

Newcastle disease virus suppresses angiogenesis in mammary adenocarcinoma models

Cancer cells heavily utilise angiogenesis process to increase vascularisation for tumour mass growth and spread, so targeting this process is important to create an effective therapy. The AMHA1 strain of Newcastle disease virus (NDV) is an RNA virus with natural oncotropism. NDV induces direct tumour cytolysis, apoptosis, and immune stimulation. This work aimed to test NDV anti-angiogenic activity in a breast cancer model. To evaluate NDV’s antitumour effect in vivo, NDV was tested against mammary adenocarcinoma AN3 transplanted in syngeneic immunocompetent mice. In vivo antiangiogenic activity was evaluated by quantifying the blood vessels in treated and control tumour sections. In vitro experiments that exposed AMN3 mammary adenocarcinoma cells and Hep-2 laryngeal carcinoma cells to NDV at different time intervals were performed to identify the exact mechanism of anti-angiogenesis by using angiogenesis microarray slides. In vivo results showed significant tumour regression and significant decrease in blood vessel formation in treated tumour sections. The in vitro microarray analysis of 14 different angiogenesis factors revealed that NDV downregulated angiopoietin-1, angiopoietin-2, and epidermal growth factor in mammary adenocarcinoma cells. However, NDV elicited a different effect on Hep-2 as represented by the downregulation of inducible protein 10, intracellular adhesion molecule-1, and basic fibroblast growth factor beta in NDV-infected tumour cells. It was found out that microarray analysis results helped interpret the in vivo data. The results suggested that the NDV oncolytic strain reduced angiogenesis by interfering with angiogenesis factors that might reduce tumour cell proliferation, infiltration, and invasion.

The expression and significance of AKR1B10 in laryngeal squamous cell carcinoma

Aldosterone reductase family 1 member B10 (AKR1B10) is a nicotinamide adenine dinucleotide phosphate (reduced coenzyme II)-dependent oxidoreductase, and its biological functions include carbonyl detoxification, hormone metabolism, osmotic adjustment, and lipid synthesis. Studies suggested that AKR1B10 is a new biomarker for cancer based on its overexpression in epithelial tumors, such as breast cancer, cervical cancer, and lung cancer. At present, studies on the expression of AKR1B10 in laryngeal cancer have not been reported. However, we found that AKR1B10 is upregulated in laryngeal carcinoma, and its expression was negatively correlated with the degree of differentiation. In addition, AKR1B10 expression was positively correlated with tumor size; lymph node metastasis; alcohol use; and Ki-67, mutant p53, and matrix metalloproteinase 2 expression. AKR1B10 was overexpressed in Hep-2 laryngeal carcinoma cells. Oleanolic acid inhibited AKR1B10 activity and expression in Hep-2 cells and suppressed Hep-2 cell proliferation, migration, and invasion. Therefore, AKR1B10 may be related to the development of laryngeal carcinoma, suggesting its use as a prognostic indicator for laryngeal cancer.

Effects of Epidermal Growth Factor Receptor Signal Pathway on Proliferation, Invasion and E-Cadherin/Vimentin Protein Expression in Laryngeal Carcinoma Hep-2 Cells

Epidermal growth factor receptor (EGFR) is one transmembrane receptor with a high expression in more than 90% of head-neck squamous carcinoma cells. This study investigated the role of EGFR signal pathway on proliferation, invasion and expression of related proteins E-cadherin/Vimentin expression in laryngeal carcinoma cells. Laryngeal carcinoma Hep-2 cells were treated with EGFR agonist EGF or inhibitor Gefitinib followed by measuring cell cycle, proliferation index (PI) and apoptosis by flow cytometry. Transwell assay was employed for cell invasion and migration. Western blotting was further employed to test E-cadherin and Vimentin level. Compared to blank control group, EGF-treated cells had significantly lower S percentage of cell cycle at 6 h, 12 h and 24 h, plus higher PI. With prolonged incubation time, S ratio and PI were further significantly elevated, with more potent cell invasion and migration abilities, lower E-cadherin and Vimentin protein levels (p < 0.05). Gefitnib significantly elevated S ratio at 6 h, 12 h and 24 h cell cycle, reduced PI, invasion or migration ability, as upregulated E-cadherin and downregulated Vimentin protein (p < 0.05). Suppressing EGFR signal pathway could inhibit proliferation of laryngeal carcinoma cells, decrease cell invasion or migration, upregulate E-cadherin and downregulate Vimentin.

Synthetic curcumin analog: inhibiting the invasion, angiogenesis, and metastasis in human laryngeal carcinoma cells via NF-kB pathway.

Laryngeal carcinoma, the most common among head and neck squamous cell carcinoma (HNSCC), induces 1% of all cancer deaths. Curcumin the active constituent of turmeric, is shown to be effective in the treatment of various cancers. In the present study, we explored the mechanistic role of bis-demethoxy curcumin analog (BDMC-A) as a chemotherapeutic agent. We investigated its inhibitory effect on invasion, angiogenesis, and metastasis in human laryngeal carcinoma (Hep-2) cells in comparison with curcumin. The effect of curcumin and BDMC-A on transcription factors (NF-κB, p65, c-Jun, c-Fos, STAT3, 5, PPAR-γ, β-catenin, COX-2, MMP-9, VEGF, TIMP-2) involved in signal transduction cascade, invasion, and angiogenesis in Hep-2 cells were quantified using Western blotting and RT-PCR technique. ELISA was used to measure the pro-inflammatory markers in Hep-2 cells treated with curcumin and BDMC-A. The results showed that BDMC-A inhibits the transcription factors NF-κB, p65, c-Jun, c-Fos, STAT3, STAT5, PPAR-γ and β-catenin, which are responsible for tumor progression and malignancy. Moreover, BDMC-A treatment downregulated MMP-9, VEGF, TGF- β, IL-6 and IL-8 and upregulated TIMP-2 levels. The effects were more significant compared to curcumin. Our overall results revealed that BDMC-A more effectively inhibited the markers of invasion, angiogenesis and metastasis in comparison with curcumin.

BODIPY and 2,3-Dihydrophthalazine-1,4-Dione Conjugates As Heavy Atom-Free Chemiluminogenic Photosensitizers.

We disclose an interesting concept for developing heavy atom-free chemiluminogenic photosensitizers. To accomplish this, conjugates 2 and 3, which are composed of boron-dipyrromethene (BODIPY) and 2,3-dihydrophthalazine-1,4-dione units, are investigated. 2 and 3 are compared in terms of their photophysical properties, chemiluminescence responses, and singlet oxygen production. Strikingly, the results indicate that decoration of BODIPY with the 2,3-dihydrophthalazine-1,4-dione scaffold boosts the singlet oxygen generation. Furthermore, treatment of epidermoid laryngeal carcinoma Hep-2 (Hep-2) cells with conjugates 2 and 3 results in efficient cellular internalization which ensures live- cell imaging of Hep-2 cells. Finally, it is noteworthy that in vitro cytotoxicity assays reveal that both 2 and 3 induce cytotoxicity when illuminated with red light. Thus, 2 and 3 represent heavy atom-free chemiluminogenic photosensitizers.

The therapeutic effects of Ficus carica extract as antioxidant and anticancer agent

Natural extracted ingredients are evolutionarily selected molecules and are used to control inflammation, and cancerous progression and transformation. Several studies have evaluated the beneficial properties of certain pharmaceutical molecules extracted from Ficus carica (FC). The current study aimed to investigate the antioxidant and anticancer properties of FC ethanolic leaves extract. The dried coarse powder of F. carica leaves was exhaustively extracted with ethanol. The resulting crude ethanolic F. carica extract (FCE) was assayed for total phenolic content and, antioxidant and anticancer activities, since both are interrelated. The antioxidant activity of the FCE was determined using DPPH as a standard stable free radical. The anticancer activity of the extract was determined against breast cancer (MCF7), hepatocellular carcinoma (HepG2), Colon cancer (CaCo-2), and a human laryngeal carcinoma (Hep-2) cell lines. The obtained results indicated a strong antioxidant activity of the extract with 1 mg/ml having 75.7% DPPH scavenging ability. The anti-cancer activities of the extract showed strong inhibition percentages against all the selected cell lines. The FCE had potent effects against both Hep2 and HepG2 cells with inhibition percentages ranged 80.7–66.9%. The properties of the FCE demonstrated here support our proposal of the validity of the traditional health claims of medicinal plants.

Abstract P-39: Fluorescent Silver Nanoclusters with Immunoglobulins and Albumins

Background: Multiplex biomedical assays including molecular genetic tests and immunoanalysis require multiple fluorophores with a wide excitation range and different emission spectra. In comparison with organic fluorophores and quantum dots, the metal nanoclusters (NC) consisting of a few to hundred atoms have the following advantages: small size, large Stokes shift, prolonged fluorescence lifetime and biocompatibility. Our research was aimed at construction of fluorescent AgNC with the main blood proteins and transmission electron microscopy (TEM). Methods: AgNC were synthesized from AgNO3 in the presence of albumins and immunoglobulins (Ig) of different classes and origin at pH 9-11 with NaBH4 recovery. The resulting AgNC with proteins were loaded to "Formvar/Carbon 200 Mesh Copper" copper grids (Ted Pella, USA) and examined using TEM system JEM 2100 Plus (JEOL, Japan) without contrast. Fluorescence excitation/emission spectra were measured in quartz cuvette using the FluoroMax + spectrofluorometer (Horiba Scientific, Japan). Results: Recovery of Ag+ ions did not occur in the presence of IgG and albumins without NaBH4 at different temperatures, pH, and incubation time. Broad excitation spectra of AgNC were in a range 340-540 nm. Their emission spectra correlated with the original AgNO3 concentration and did not depend on protein and pH. NC stabilized with IgG or albumin with blue fluorescence and emission maximum at 420 nm contained NC from 0.6 nm and higher. Green AgNC with proteins had bright fluorescence at 430-470 nm and red NC showed emission maximum at 650 nm. TEM revealed discrete AgNC and their numerous aggregates in each sample of fluorescent NC in spite of different fluorescent emission spectra. According to the MTT test, AgNC with human IgG and BSA with protein concentrations up to 3 mg/ml were not toxic for human larynx carcinoma HEp-2 cells despite cytotoxicity of silver nanoparticles covered with IgG or albumin envelopes as well as Cd and AuNC with BSA. Conclusion: AgNC with antibodies and albumin with a broad size range and aggregation possess tunable fluorescence emission spectra with broad excitation at 340-540 nm. Different emission spectra permit AgNC to be used in multiplex assays. AgNC were not toxic for human tissue culture and may be applied for bioimaging.

Efficacy of nimotuzumab (hR3) conjugated with 131I or 90Y in laryngeal carcinoma xenograft mouse model

Purpose The humanized monoclonal antibody hR3, both alone and in combination with other chemotherapeutic agents and radiotherapy, can be used to treat head and neck cancers. Substantial progress has been made in the development of targeted radioimmunotherapy using iodine-131 (131I) and yttrium-90 (90Y) radioisotopes in recent years. In the present study, we examined the efficacy of hR3 conjugated with 131I or 90Y to inhibit tumor growth in a laryngeal carcinoma xenograft tumor model. Methods hR3 was labeled with 131I or 90Y to generate the conjugates 131I-hR3 or 90Y-hR3. The conjugates were incubated with HEp-2 laryngeal carcinoma cells to evaluate binding capacity. The efficacy of the labeled hR3 conjugates to treat laryngeal cancer was also evaluated in nude mice inoculated with HEp-2 tumors. Results The purified radioimmunoconjugates with specific activities of 187–191 MBq/mg had radiochemical purity >98% and >80% immunoreactivity with HEp-2 cells. Mice with HEp-2 xenografts treated with 131I-hR3 or 90Y-hR3 showed reduced tumor volume and improved survival rates compared to the untreated control group and the group treated with unlabeled hR3. At equivalent doses, radioimmunotherapeutic hR3 labeled with 90Y had increased tumor inhibition activity compared to hR3 labeled with 131I. Conclusions 131I-hR3 and 90Y-hR3 are promising targeted radiopharmaceuticals for treatment of head and neck cancers, especially laryngeal cancers.

PH-Sensitive Dairy-Derived Hydrogels with a Prolonged Drug Release Profile for Cancer Treatment.

A novel versatile biocompatible hydrogel of whey protein isolate (WPI) and two types of tannic acid (TAs) was prepared by crosslinking of WPI with TAs in a one-step method at high temperature for 30 min. WPI is one common protein-based preparation which is used for hydrogel formation. The obtained WPI-TA hydrogels were in disc form and retained their integrity after sterilization by autoclaving. Two TA preparations of differing molecular weight and chemical structure were compared, namely a polygalloyl glucose-rich extract-ALSOK 02-and a polygalloyl quinic acid-rich extract-ALSOK 04. Hydrogel formation was observed for WPI solutions containing both preparations. The swelling characteristics of hydrogels were investigated at room temperature at different pH values, namely 5, 7, and 9. The swelling ability of hydrogels was independent of the chemical structure of the added TAs. A trend of decrease of mass increase (MI) in hydrogels was observed with an increase in the TA/WPI ratio compared to the control WPI hydrogel without TA. This dependence (a MI decrease-TA/WPI ratio) was observed for hydrogels with different types of TA both in neutral and acidic conditions (pH 5.7). Under alkaline conditions (pH 9), negative values of swelling were observed for all hydrogels with a high content of TAs and were accompanied by a significant release of TAs from the hydrogel network. Our studies have shown that the release of TA from hydrogels containing ALSOK04 is higher than from hydrogels containing ALSOK 02. Moreover, the addition of TAs, which display a strong anti-cancer effect, increases the cytotoxicity of WPI-TAs hydrogels against the Hep-2 human laryngeal squamous carcinoma (Hep-2 cells) cell line. Thus, WPI-TA hydrogels with prolonged drug release properties and cytotoxicity effect can be used as anti-cancer scaffolds.

Investigation of the Functional Role of mir-1825 in Laryngeal Squamous Cell Carcinoma Hep-2 Cell Line

Investigation of the Functional Role of mir-1825 in Laryngeal Squamous Cell Carcinoma Hep-2 Cell Line

L-ascorbic acid induces apoptosis in human laryngeal epidermoid Hep-2 cells by modulating the nuclear factor kappa-light-chain-enhancer of activated B cells/ mitogen-activated protein kinase/Akt signaling pathway

L-ascorbic acid (L-AA; vitamin C) induces apoptosis in cancer cells. This study aimed to elucidate the molecular mechanisms of L-AA-induced apoptosis in human laryngeal epidermoid carcinoma Hep-2 cells. L-AA suppressed the viability of Hep-2 cells and induced apoptosis, as shown by the cleavage and condensation of nuclear chromatin and increased number of Annexin V-positive cells. L-AA decreased Bcl-2 protein expression but upregulated Bax protein levels. In addition, cytochrome c release from the mitochondria into the cytosol and activation of caspase-9, -8, and -3 were enhanced by L-AA treatment. Furthermore, apoptosis-inducing factor (AIF) and endonuclease G (EndoG) were translocated into the nucleus during apoptosis of L-AA-treated Hep-2 cells. L-AA effectively inhibited the constitutive nuclear factor-κB (NF-κB) activation and attenuated the nuclear expression of the p65 subunit of NF-κB. Interestingly, L-AA treatment of Hep-2 cells markedly activated Akt and mitogen-activated protein kinase (MAPK; extracellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase [JNK]) and and LY294002 (Akt inhibitor), SB203580 (p38 inhibitor) or SP600125 (a JNK inhibitor) decreased the levels of Annexin V-positive cells. These results suggested that L-AA induces the apoptosis of Hep-2 cells via the nuclear translocation of AIF and EndoG by modulating the Bcl- 2 family and MAPK/Akt signaling pathways.