Acridine Orange/ethidium Bromide Research Articles

Targeted silencing of the MCL-1 gene using multi-layered dendrimer-based nanoconstructs achieves efficient tumor regression in xenografted mice models

The setback in the practical clinical use of RNA interference (RNAi)-based cancer treatment stems from the lack of targeted small interfering RNA (siRNA) delivery. Here, we show that luteinizing hormone-releasing hormone(LHRH) analog-tethered multi-layered polyamidoamine (PAMAM) nanoconstructs silence the anti-apoptotic MCL-1 gene in LHRH receptor overexpressing human breast (MCF-7) and prostate cancer (LNCaP) cells with 70.91 % and 74.10 % efficiency, respectively. These results were confirmed by RT-PCR. The Acridine orange/Ethidium bromide (AO/EB) dual staining revealed that the silencing of MCL-1 induced apoptosis in both the cell lines. In vivo tumor regression studies performed using MCF-7 and LNCaP xenografted severe combined immunodeficiency(SCID) mice demonstrated highly improved tumor regression in groups treated with targeted nanoconstructs complexed with MCL-1 siRNA (T + siMCL-1) compared to the other treatment groups. The quantitative RT-PCR results of tumor tissues demonstrated significant MCL-1 gene silencing, i.e., 73.76 % and 92.63 % in breast and prostate tumors, respectively, after T + siMCL-1 treatment. Reduction in MCL-1 protein expression as assessed by immunohistochemistry further confirmed these results. Furthermore, the caspase 3/7 assay demonstrated apoptosis in the MCL-1 silenced tissues. The study strongly suggests that targeted delivery of siRNAs using multi-layered dendrimer nanostructures could be an effective therapy for LHRH overexpressing cancers.

Acacia Honey-derived Bioactive Compounds Exhibit Induction of p53-dependent Apoptosis in the MCF-7 Human Breast Cancer Cell Line

Background Research studies have focused on discovering new anti-proliferative and pro-apoptotic agents derived from natural products from which honey constitutes a prominent candidate. The Acacia honey (AH) is known to display anticancer activity, but the mechanisms of action are still not well defined. Objectives Using in vitro and computational approaches, we aimed to assess the interaction among selected bioactive compounds derived from AH, with the apoptotic protein p53, which could trigger apoptosis. Materials and Methods The phytocompounds of AH were investigated via gas chromatography–mass spectrophotometry analysis. The cytotoxic effect and induced apoptosis on the MCF-7 breast cancer cell line were assessed by 3-(4,5-dimethylthiazolyl-2)-2,5 diphenyltetrazolium bromide and acridine orange-ethidium bromide staining approaches. The molecular docking analysis between AH compounds and p53 was carried out. Results The drug-likeness prediction revealed that most of the identified compounds meet Lipinski’s rules. We demonstrate that AH exerts an interesting cytotoxic effect in a dose-dependent manner against the MCF-7 cell line with IC50 5.053µg/mL. Significant cell alterations and notable induced apoptosis were detected when cells were treated with AH. The molecular docking analysis revealed that melezitose is among the most important potential bioactive compounds that interact with p53 leading to apoptosis. The binding affinity was −8.1 kcal/mol, and the closest molecular interactions in the active site included 10 residues, which could explain the potential biological activity. Conclusion This work sheds light on AH as a significant source of bioactive chemicals with potential for promoting apoptosis that may be exploited as an alternative therapy for breast cancer.

Oroxylum indicum Vent Root Bark Extract Inhibits the Proliferation of Cancer Cells and Induce Apoptotic Cell Death

Oroxylum indicum Vent is a medium-sized deciduous tree that belongs to the family Bignoniaceae. The roots of this tree are used as one of the ten ingredients to prepare the Dasamula formulation in the Ayurvedic system of medicine in India. Although traditional uses are attributed to the medicinal plant, there are limited scientific data on its potential. The present study thus analyzed the cytotoxic and apoptotic potential of the plant against different cancer cells. MTT assay was used to determine cytotoxicity using HeLa, HCT 15, and MDA-MB-231 cells, with the IC50 values, revealed at concentrations of 92.43, 133.0, and 112.84 µg/mL respectively. However, the extract was less toxic to non-cancer cells. HeLa cells further treated with OIM were subjected to flow cytometric analysis for studying the cell cycle stages. When untreated cells at G1 phase were found at a relative percentage of 71.9%, it increased to 79.3 and 86% with OIM treatment at concentrations of 50 and 100 µg/mL; cells in the S phase decreased from 10.3 to 8.2 and 7.5%, concluding the arrest of the cell cycle at G1 phase. With further study of apoptotic morphology with dual acridine orange–ethidium bromide staining and Annexin–Hoechst staining, cells at early and late apoptotic stages were observed with OIM treatment at 100 µg/mL concentration. Although such effects were noticed with OIM treatment, it could not be concluded that the extract had remarkable anti-proliferative effects, since the small changes noticed in cell cycle arrest and apoptotic induction were attained at a high concentration of OIM 100 µg/mL. The biological activities of plants and their extracts are attributed to the presence of multifarious compounds present in them. LC-MS Q-TOF analysis confirmed the presence of biochanin A and baicalein in OIM. HPLC-based quantification of baicalein and chrysin was shown to be 3.36 and 1.11 mg/gram dry weight. To conclude, the above results suggest that the root bark of O. indicum has a broad spectrum of biological activities, including anticancer and apoptotic properties.

In vitro anti-prostate adenocarcinoma and lung cancer studies of phenoxyaniline-block-poly(methyl methacrylate) based nanocomposites via controlled radical polymerization.

A phenoxyaniline-based macroinitiator is utilized for the first time in order to produce phenoxyaniline-block-poly(methyl methacrylate) composites through single electron transfer-living radical polymerization (SET-LRP) under mild conditions. A different weight percentage of Cloisite 93A is added into the polymer mixtures in order to increase their biochemical properties. The prepared block copolymer nanocomposites are characterized using ATR-IR, UV-vis-spectroscopy, XRD, Raman, TGA, DSC, a particle size analyzer, contact angle measurements and SEM in order to characterize their structural, thermal, surface and morphological properties. Further, the developed polymeric nanocomposites are successfully applied in two different cancer cell lines (prostate adenocarcinoma and lung cancer), which show excellent anticancer properties. Also, acridine orange/ethidium bromide (AO/EtBr) dual staining is performed, which causes drastic cell death by apoptosis in both A549 and PC-3 cell lines, which indicated that the prepared polymeric nanocomposites effectively inhibit the cell proliferation and induce the apoptosis in both the cancer cells. Here nanoclay is used for cancer treatment because of its complete water solubility, which essentially causes the formation of a cationic complex between the clay and drug through electrostatic interactions. Hence, the exchange of ions between the clay and other ions in the biological environment leads to inhibition of the proliferation of prostate adenocarcinoma and lung cancer cells in the system.

Apoptosis-Mediated Anticancer Activity of Ganoderma colossus (Agaricomycetes) Extracts in Breast Cancer Cells.

Cancer is a leading cause of death worldwide. The current cancer treatments including chemo-, radio- and immuno-therapies pose various side effects, and chances of recurrence that demand for new therapeutics to overcome the issues with existing ones. Mushrooms are considered a potential source of novel therapeutic agents. Ganoderma colossus, a non-edible wood-inhabiting mushroom, is known for certain medical properties. The present study aimed to investigate the possible anticancer activity of methanolic, ethyl acetate, and chloroform extracts of G. colossus, against MCF-7 cells and the mechanism of action(s). MTT assay and gene expression studies were carried out by following the standard protocols. The results demonstrated that among the three solvents, the ethyl acetate crude extract of the mushroom exhibited potential cytotoxic activity on MCF-7 (IC50, 17.2 ± 2.7). The DNA damage induced by the solvent extracts of G. colossus was observed by H2AX foci formation. The TP53 over-expression and flow cytometry analysis indicated that checkpoint activation followed by cell cycle arrest occurred at G1/G0 phase in response to the extract treatment. The dual acridine orange/ethidium bromide (AO/EB) staining revealed apoptosis-associated changes in the cells. Analysis of caspase 3 activations by immunophenotyping confirmed the apoptotic process in the extract-treated cells. Bcl-2 and TP53 mRNA expression data by RT-PCR disclosed the apoptosis pathway. The GC- MS spectral data of the ethyl acetate crude extract of the mushroom indicated the presence of molecules capable of inducing apoptosis. The present study warrants further studies to isolate the molecule(s) from G. colossus which may be a potential drug candidate for breast cancers.

Hydroalcoholic extract of Taraxacum officinale induces apoptosis and autophagy in 4T1 breast cancer cells.

Triple-negative breast cancer (TNBC) is an aggressive and deadly breast cancer sub-type with limited therapeutic options. Dandelion (Taraxacum officinale) exhibiting extensive anti-cancer activity is reported to be effective against TNBC; however, its anti-tumor effect mechanisms have not been fully elucidated. The purpose of this study was to determine the anti-cancer activity of hydroalcoholic extract of dandelion (HADE) on 4T1 cells, and the mechanism of HADE-induced cell death. The effect of HADE on cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assays. Apoptotic cell death was monitored by flow cytometry. The DNA fragmentation was evaluated by Acridine orange/Ethidium bromide (AO/EB) staining. Nitric oxide (NO) level was detected using Griess assay. The effects of HADE on Atg-7, Beclin-1, Bcl2, Bax and p53 genes were investigated by real-time reverse transcription-polymerase chain reaction. The results showed that HADE inhibited cell growth and proliferation in a dose- and time-dependent manner. The HADE induced 4T1 breast cancer cell death via apoptosis and autophagy. The DNA fragmentation was improved as the concentration of HADE increased. The NO secretion was declined with increasing concentration of HADE. Gene expression analysis confirmed HADE-induced apoptosis and autophagy in cancer cells. The Bax, Bax/Bcl-2 ratio, p53, Beclin-1 and Atg-7 over-expression as well as Bcl-2 down-regulation were also evident in treated cancer cells.

Anticancer Activity of the Bioactive Extract of the Morel Mushroom (Morchella elata, Ascomycetes) from Kashmir Himalaya (India) and Identification of Major Bioactive Compounds.

Morel mushrooms, Morchella species are highly nutritional, excellently edible and medicinal. Anticancer activity of M. elata, growing in forests of Kashmir Himalaya was studied. Ethyl acetate extract of fruiting bodies of M. elata (MEAE) was evaluated for cytotoxicity by MTT assay using Daltons lymphoma ascites (DLA), human colon cancer (HCT-116) and normal cell lines. Anti-carcinogenic and antiangiogenic activities of MEAE were tested using mouse models. Proapoptotic activity was detected by double staining of acridine orange-ethidium bromide assay. MEAE was partially purified by column chromatography and the bioactive compounds were identified by LC-MS analysis. The bioactive extract of M. elata showed significant cytotoxicic activity against DLA (P < 0.05), HCT-116 cell lines (P < 0.05) and did not possess appreciable adverse effect on the viability of normal cells. At a concentration of 100 µg/mL, 60% cell death was observed in HCT-116 cell line while 80% cell death was found in DLA cell line. The extract also possessed profound anticarcinogenic, antiangiogenic and proapoptotic activities. LC-MS analysis showed celastrol (RT 9.504, C29H38O4, MW 450.27), convallatoxin (RT 9.60, C29H42O10, MW 550.27), cucurbitacin A (RT 11.97, C32H46O9, MW 574.71) and madecassic acid (RT 14.35, C30H48O6, MW 504.70) as the major bioactive components. Current experimental studies indicated that bioactive extract of M. elata possessed significant anticancer activity. Being an excellently edible mushroom, the potential therapeutic use of M. elata and its bioactive extract in complementary therapy of cancer is envisaged.

Evaluation of antimicrobial and antioxidant activity of cleome gynandra l. and assess its anticancer activity against in vitro cancer model

Cleome gynandra Linn. is the native herb distributed at tropical and sub-tropical temperature. In this study, the antioxidant, antimicrobial and anticancer properties of the ethanolic extract of Cleome gynandra leaves were investigated also the anti-cancer activity was assessed against A549 lung cancer cell lines. The antioxidant activities were evaluated by 1,1‑diphenyl‑2‑picrylhydrazyl free radical scavenging activity. The IC50 value of DPPH was ranged from 65.67 µg/ml for the leaf extract. The anti-microbial activity was showed against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia and Salomonella typhi. The cytotoxic study was conducted and the results exhibited the strongest inhibitory activity against A549 lung cancer cell line as concentration dependent manner. The cytotoxic activity of the leaf extract was supported by the induction of apoptosis cell. The other assays to evaluate anticancer activity are DAPI (4,6 – diamminodino-2-phenylindole), PI (propidium iodide) and EtBr/AO (Ethidium bromide/acridine orange) staining methods. All the study results revealed the efficient anticancer activity of the ethanolic leaf extract of Cleome gynandra leaves. Thus, the herbal drug treatment might highly be recommended to treat effectively lung cancer cell line as an ideal choice or combinational therapy.

The Essential Oil from Oliveria decumbens Vent. (Apiaceae) as Inhibitor of Breast Cancer Cell (MCF-7) Growth.

Oliveria decumbens Vent. is an aromatic and medicinal plant traditionally used in Iran for the treatment of infections, gastrointestinal diseases, cancer, and inflammation. This research was aimed at investigating the pharmacological potential of O. decumbens essential oil (OEO) and its main compounds, focusing on OEO's cytotoxic effects on MCF-7 breast cancer cells. OEO was obtained by hydro-distillation, and the chemical constituents were identified using GC-MS. Thymol, carvacrol, γ-terpinene, and p-cymene were the main OEO constituents. When MCF-7 cells were treated with OEO, the expressions of genes related to apoptosis (BIM and Bcl-2), tumor suppression (PTEN), and cell growth inhibition (AURKA), were evaluated using real-time PCR. Moreover, molecular docking was used for studying in silico the interaction of OEO principal compounds with PTEN and AURKA. The expression of AURKA was significantly reduced since the OEO treatment enhanced the expression of PTEN. Through in silico molecular docking, it was revealed that thymol, carvacrol, p-cymene, and γ-terpinene can activate PTEN and thus inhibit AURKA. Additionally, the DNA fragmentation assay, acridine orange/ethidium bromide (AO/EB) double-staining assay, and real-time PCR highlighted the fact that the OEO treatment could activate apoptosis and inhibit cell proliferation. Therefore, OEO is a viable candidate to be employed in the pharmaceutical industry, specifically as a possible agent for cancer therapy.

Enhancement of cytotoxic and antioxidant activities of Digenea simplex chloroform extract using the nanosuspension technique

Digenea simplex (D. simplex), an Egyptian marine red macroalga, contains a diverse group of phytochemicals with unique bioactivities. At the same time, the synthesis of nanosuspension (NS) has received increasing interest to optimize the technological aspects of drugs. Thence, the main objective of this work was to use the chloroform extract (ChlE) of D. simplex to prepare its nanosuspension (ChlE-NS) formulation to increase its aqueous solubility, thereby improving its bioactivity. By using FTIR, GC/MS analysis, and phytochemical screening assays, the chemical profiling of ChlE was assessed. NS was prepared by the antisolvent precipitation technique using 1.5% w/v polyvinyl alcohol (PVA). A light microscope, FTIR, particle size distribution, polydispersity index (PDI), and zeta potential (ZP) measurements was used to characterize the prepared NS. Four cancer cell lines were used in the MTT experiment to investigate the anticancer potential of ChlE and ChlE-NS. An apoptotic mechanism was established using acridine orange/ethidium bromide (AO/EB) dual staining, DNA fragmentation, and increased caspase activity. ChlE and ChlE-NS were also evaluated as antioxidants using DPPH and ABTS free radical assays. The results showed that, when compared to ChlE, ChlE-NS had greater cytotoxic activity against the four cancer cell lines. However, results of antioxidant activity showed that ChlE-NS had an IC50 of 36.86 ± 0.09 and 63.5 ± 0.47%, while ChlE had values of 39.90 ± 0.08 and 86.5 ± 0.8% in DPPH and ABTS assays, respectively. Based on the results of this research, D. simplex ChlE-NS may be an effective strategy for enhancing ChlE's cytotoxic and antioxidant activities.Graphical abstract

Synthesis of Quinoline-2-Carboxylic Acid Aryl Ester and Its Apoptotic Action on PC3 Prostate Cancer Cell Line.

The aim of the present study is to synthesise an aryl ester compound from quinoline-2-carboxylic acid to evaluate its apoptotic, cell cycle blockade, and antiproliferative activity on the prostate cancer cell lines (PC3). Chromatographic and spectroscopic analysis was used to identify the synthesised carboxylic acid compound. The synthesised compound was treated with a PC3 cell line for 24h with control. The cells were treated at various concentration ranges of 0, 3.91, 7.81, 15.63, 31.25, 62.5, 125, 250, 500, and 1000µg/mL each. The cytotoxicity effect was studied by MTT assay, and their anticancer activity was further evaluated using cell cycle analysis, DNA fragmentation assay, acridine orange-ethidium bromide staining, and Western blot analysis. The end antiproliferative result showed that PC3 cell viability decreases in a concentration-dependent manner and the synthesised compound exhibited potent cytotoxicity against PC3 cells with an IC50 value of 26µg/mL at the concentration of 125µg. The increase in the number of apoptotic cells was observed after treating PC3 cells with the sample in double-staining methods. S phase of the cell cycle was significantly blocked by the test sample, and a typical ladder pattern of internucleosomal fragmentation was observed. A decrease in the live cells was observed with the sample in AO/ET-BR. A significant increase in the Bax expression and a decrease in Bcl-2 expression observed enhance the activity of caspases-7 and -9. The synthesised compound had shown to possess excellent cytotoxic effect through inducing apoptosis, especially causing cell cycle arrest at the S phase.

Synthesis, spectral characterization, protein binding and cytotoxic evaluation of new cobalt(II) and cobalt(III) complexes containing benzimidazolylterpyridine as ligand

Two new six-coordinate cobalt(II) [Co(bitpy)2]Cl2 (1) and cobalt(III) [Co(bitpy)2](ClO4)3 (2) complexes containing benzimidazolylterpyridine (bitpy) ligand have been synthesized and characterized using spectroscopic and electrochemical techniques. The g ‖ and g ⊥ values in the EPR spectrum for 1 were 2.215 and 2.012, respectively, and clearly authenticated the formation of stable octahedral low spin Co(II) meridional isomer complex. Similarly, 1H NMR spectrum of 2 revealed low spin and its diamagnetic nature. The interaction of the complexes with bovine serum albumin (BSA) has been investigated and revealed that the cobalt complexes quenched the intrinsic fluorescence by following the static quenching mechanism. MTT assay was used to check the anticancer potential of 2 against human breast cancer cells (MCF-7) and human cervical cancer cells (HeLa). The IC50 values of 2 were lower than 7 µM in both the cancer cells and higher than 100 µM for non-cancerous cells (human embryonic kidney cells), indicating the specificity of the complex toward cancer cells. Furthermore, the morphological changes induced by 2 on the tested cell lines were monitored by AO-EB (acridine orange-ethidium bromide) and DAPI (4′,6-diamidino-2-phenylindole) staining methods.

Quality by design steered development of Niclosamide loaded liposomal thermogel for Melanoma: In vitro and Ex vivo evaluation

Melanoma is the most malignant form of skin cancer across the globe. Conventional therapies are currently ineffective which could be attributed to the rampant chemo-resistance, metastasis, inability to cross the skin barriers and accumulate within the tumor microenvironment. This advent brings in the principles of drug repurposing by repositioning Niclosamide (NIC), an anthelmintic drug for skin cancer. Incorporation into the liposomes facilitated enhanced melanoma cell uptake and apoptosis. Cytotoxicity studies revealed 1.756-fold enhancement in SK-MEL-28 cytotoxicity by NIC-loaded liposomes compared to free drug. Qualitative and quantitative cell internalization indicated greater drug uptake within the melanoma cells illustrating the efficacy of liposomes as efficient carrier systems. Nuclear staining showed blebbing and membrane shrinkage. Elevated ROS levels and apoptosis shown by DCFDA and acridine orange-ethidium bromide staining revealed greater melanoma cell death by liposomes compared to free drug. Incorporating NIC liposomes into the thermogel system restricted the liposomes as a depot onto the upper skin layers. Sustained zero order release up to 48 h with liposomes and 23.58-fold increase in viscosity led to the sol-to-gel transition at 33℃ was observed with liposomal thermogel. Ex vivo gel permeation studies revealed that C-6 loaded liposomes incorporated within the thermogel successfully formed a depot over the upper skin layer for 6 h to prevent transdermal delivery and systemic adverse effects. Thus, it could be concluded that NIC loaded liposomal thermogel system could be an efficacious therapeutic alternative for the management of melanoma.

Ilimaquinone (Marine Sponge Metabolite) Induces Apoptosis in HCT-116 Human Colorectal Carcinoma Cells via Mitochondrial-Mediated Apoptosis Pathway.

Ilimaquinone (IQ), a metabolite found in marine sponges, has been reported to have a number of biological properties, including potential anticancer activity against colon cancer. However, no clear understanding of the precise mechanism involved is known. The aim of this study was to examine the molecular mechanism by which IQ acts on HCT-116 cells. The anticancer activity of IQ was investigated by means of a cell viability assay followed by the determination of induction of apoptosis by means of the use of acridine orange–ethidium bromide (AO/EB) staining, Annexin V/PI double staining, DNA fragmentation assays, and TUNEL assays. The mitochondrial membrane potential (ΔΨm) was detected using the JC-1 staining technique, and the apoptosis-associated proteins were analyzed using real-time qRT-PCR. A molecular docking study of IQ with apoptosis-associated proteins was also conducted in order to assess the interaction between IQ and them. Our results suggest that IQ significantly suppressed the viability of HCT-116 cells in a dose-dependent manner. Fluorescent microscopy, flow cytometry, DNA fragmentation and the TUNEL assay in treated cells demonstrated apoptotic death mode. As an additional confirmation of apoptosis, the increased level of caspase-3 and caspase-9 expression and the downregulation of Bcl-2 and mitochondrial dysfunction were observed in HCT-116 cells after treatment with IQ, which was accompanied by a decrease in mitochondrial membrane potential (ΔΨm). Overall, the results of our studies demonstrate that IQ could trigger mitochondria-mediated apoptosis as demonstrated by a decrease in ΔΨm, activation of caspase-9/-3, damage of DNA and a decrease in the proportion of Bcl-2 through the mitochondrial-mediated apoptosis pathway.

Di-(2-Ethylhexyl) Phthalate and Microplastics Induced Neuronal Apoptosis through the PI3K/AKT Pathway and Mitochondrial Dysfunction.

Di-(2-Ethylhexyl) phthalate (DEHP) and microplastics (MPs) have released widespread residues to the environment and possess the ability to cause damage to humans and animals. However, there are still gaps in the study of damage to neurons caused by DEHP and MPs in mice cerebra and whether they have combined toxic effects. To investigate the underlying mechanism of action, mice were fed 200 mg/kg DEHP and 10 mg/L MPs in vivo. In vitro, NS20Y (CBNumber: CB15474825) cells were treated with 25 μM DEHP and 775 mg/L MPs. Next, qRT-PCR and western blot analysis were performed to evaluate PI3K/AKT pathway genes, mitochondrial dynamics-related genes, apoptosis-related genes, and GSK-3β and its associated genes, mRNA, and protein expression. To determine pathological changes in the mice cerebra, hematoxylin and eosin (H&E) staining, transmission electron microscopy, and TUNEL staining were employed. To determine the levels of reactive oxygen species (ROS) and apoptosis cells in vitro, ROS staining, acridine orange/ethidium bromide (AO/EB) staining, and flow cytometry were performed. Our results demonstrated that DEHP and MPs caused changes in mitochondrial function, and GSK-3β and its associated gene expression in mice through the PI3K/AKT pathway, which eventually led to apoptosis of neurons. Moreover, our findings showed that DEHP and MPs have a combined toxic effect on mice cerebra. Our findings facilitate the understanding of the neurotoxic effects of DEHP and MPs on neurons in the cerebra of mice and help identify the important role of maintaining normal mitochondrial function in protecting cerebrum health.

Cross-interactions between Norepinephrine, Methicillin-Resistant Staphylococcus aureus and Human Osteoblast Cells in Culture Conditions

Objective: The role of norepinephrine (NE) on growth, adhesion and invasion of methicillin-resistant Staphylococcus aureus (MRSA) ATTC 43300 was examined in human osteoblast (HOB) cells. The effects of NE and/or MRSA on the viability and cell death pathways of HOB cells were also investigated. Furthermore, the alterations of bacterial response to oxidative stress (H2O2) were analyzed in the presence/ absence of NE. Materials and Methods: Bacterial growth was detected spectrophotometrically. The colony counting method was examined for adhesion-invasion. The alteration of HOB cell viability was determined by methyl thiazolyl diphenyl-tetrazolium bromide (MTT) assay. The death pathways of HOB cells were examined microscopically using acridine orange-ethidium bromide dual staining and dichlorofluorescein-diacetate (DCF-DA) dye. The bacterial response to H2O2 was investigated by agar dilution. Results: The growth of bacterium was not affected in the presence of NE. Bacterial adhesion was decreased by NE (p&lt;0.0001) while high-level NE induced invasion (p=0.013). HOB cell viability was reduced by MRSA and/or NE (p&lt;0.001). MRSA and co-existence of MRSA and NE caused necrosis more than apoptosis in HOB cells (p&lt;0.05). NE did not alter the bacterial response to oxidative stress. Conclusion: Norepinephrine has different effects on the biological properties of both MRSA and HOB cells.

Pomolic acid inhibits proliferation of human lung carcinoma cells via induction of apoptosis and suppression of cell migration and invasion

Purpose: To investigate the anti-proliferative effect of pomolic acid on lung cancer cells (A549), and theunderlying mechanism.Methods: The viability of pomolic acid-treated A549 cells was determined by MTT and colony formation assays. Cell colony formation was monitored with acridine orange/ethidium bromide (AO/EB) staining. Protein expressions of Bax and Bcl-2 were assayed by western blotting.Results: Pomolic acid suppressed the growth of A549 cells, with an half-maximal inhibitory concentration of (IC50) of 10 μM (p &lt; 0.05). However, the IC50 of pomolic acid for normal BEAS-2B cells was 80 μM. Pomolic acid also decreased colony formation of A549 cells. At 20 μM, the percentage of A549 colonies decreased to 14 % of control. The dose-dependent cytotoxicity of pomolic acid against A549 cells was mediated via induction of apoptosis and oxidative stress. Pomolic acid treatment enhanced the expression of Bax and decreased the expression of Bcl-2 in A549 cells. Moreover, pomolic acid inhibited the migration and invasion in A549 cells in a dose-dependent manner (p &lt; 0.05).Conclusion: These results indicate the potent anticancer effect of pomolic acid against human lung cancer cells. Thus, pomolic acid has promising potential as a lead molecule for the development of chemotherapy.

5-Hydroxyisophthalic acid and neocuproine containing copper(II) complex as a promising cytotoxic agent: Structure elucidation, topology, Hirshfeld surface, DFT calculations, and molecular docking analysis

We aimed to synthesize a new copper(II) complex, namely [Cu(H2IPA)(NC)(Cl)]⋅H2O (1) (where, H3IPA = 5-hydroxy isophthalic acid and NC = 2,9-dimethyl-1,10-phenanthroline or Neocuproine) as a promising chemotherapeutic agent. Complex 1 was structurally characterized by single-crystal X-ray diffraction (SCXRD), UV-Visible, Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and powder X-ray diffraction (PXRD) methods. Single crystal diffraction analysis reveals that Cu(II) ion exists in a distorted square pyramidal geometry, with the ligation of a chloride ion, two oxygen atoms, and two nitrogen atoms. Topological structural simplification of complex 1 shows ins as an underlying net. The prominent intermolecular interactions and their quantitative contributions in complex 1 were investigated using Hirshfeld Surface (HS) analysis. The DFT calculations were also performed to validate the experimental results (XRD, FTIR, and UV-Visible). In vitro cytotoxicity of complex 1 was performed against acute myeloid leukemia (THP-1), colorectal (SW480), and prostate cancer (PC-3) cell lines by utilizing MTT assay. The result shows that complex 1 has the ability to inhibit the growth of cancer cells at a lower concentration. The morphological changes that occur during apoptosis were carried out with acridine orange/ethidium bromide (AO/EB) staining methods on THP-1 and SW480 cancer cell lines. Additionally, comprehensive molecular docking studies were performed to understand the potential binding interactions of complex 1 with target protein and DNA double helix.

Galangin/β-Cyclodextrin Inclusion Complex as a Drug-Delivery System for Improved Solubility and Biocompatibility in Breast Cancer Treatment.

The purpose of this study was to evaluate the potential of a newly modified cyclodextrin derivative, water-soluble β-cyclodextrin–epichlorohydrin (β-CD), as an effective drug carrier to enhance the poor solubility and bioavailability of galangin (GAL), a poorly water-soluble model drug. In this regard, inclusion complexes of GAL/β-CDP were prepared. UV-VIS spectrophotometry, Fourier-transform infrared spectroscopy (FTIR), X-ray crystallography (XRD), zeta potential analysis, particle size analysis, field emission scanning electron microscopy (FESEM), and transmission electron microscopy (TEM) were applied to characterize the synthesized GAL/β-CD. Michigan Cancer Foundation-7 (MCF-7; human breast cancer cells) and rat embryo fibroblast (REF; normal cells) were employed to examine the in vitro cytotoxic effects of GAL/β-CD using various parameters. The dye-based tests of MTT and crystal violet clearly exhibited that GAL/β-CD-treated cells had a reduced proliferation rate, an influence that was not found in the normal cell line. The cells’ death was found to follow apoptotic mechanisms, as revealed by the dye-based test of acridine orange/ethidium bromide (AO/EtBr), with the involvement of the mitochondria via caspase-3-mediated events, as manifested by the Rh 123 test. We also included a mouse model to examine possible in vivo toxic effects of GAL/β-CD. It appears that the inclusion complex does not have a significant influence on normal cells, as indicated by serum levels of kidney and liver enzymatic markers, as well as thymic and splenic mass indices. A similar conclusion was reached on the histological level, as manifested by the absence of pathological alterations in the liver, kidney, thymus, spleen, heart, and lung.

Radiosensitizing effect of dendrosomal nanoformulation of curcumin on cancer cells.

Curcumin was found to possess numerous pharmacological activities in clinical research, however, its biological effects together with radiation are yet to be addressed. The present study investigated whether the combined treatment of dendrosomal nanoformulation of curcumin (DNC) and gamma radiation can enhance the radiosensitivity of U87MG and MDA-MB-231 cell lines. U87MG and MDA-MB-231 cell lines were exposed to 2 Gray (Gy) and 10μM DNC determined by MTT assay, then subjected to clonogenic assay, cell cycle assay, and flow cytometric apoptosis analysis. Acridine Orange/Ethidium Bromide (AO/EB) and 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) stained cells were used to study morphologic changes. The expression evaluation of putative cell cycle genes, i.e., P53, P21, CCND1, and CCNB1 was carried out by RT-qPCR. Our findings indicated that the combined treatment with DNC and radiation might cooperatively augment the efficacy of ionizing radiation in the cancer cells and notably decrease the survival and viability of the cells in a time- and concentration-dependent manner. In addition to a synergistic effect deducted by sensitizer enhancement ratio (SER) assessment, co-treatment resulted in greater apoptotic cells than the individual treatments. Further experiments then indicated that DNC could effectively induce G2/M phase cell cycle arrest and apoptosis following irradiation. Conformably, there was a decrement of CCND1 and CCNB1 expression, and an increment of P53, P21 expression. The data implied that DNC as a radiosensitizer can enhance the lethal effect of ionizing radiation on cancer cells which could be a promising adjuvant therapy in clinical treatments.