Cytotoxicity of Goniothalamin on the Human Hepatocellular Carcinoma HEPG2 Cell Line

Goniothalamin is a biologically active styrylpyrone derivative isolated from various Goniothalamus sp., belonging to the Annonacae family. This plant extract has been reported to be cytotoxic towards several tumor cell lines such as pancreas carcinoma (PANC-1), gastric carcinoma (HGC-27) and breast carcinoma (MCF-7). The purpose of this study was to examine and characterize the in vitro cytotoxicity effect of goniothalamin on the human hepatocellular carcinoma HepG2 cells and normal liver Chang cells and also to study the morphological and biochemical changes of goniothalamin-treated HepG2 and Chang cells. Goniothalamin (2.3 -150 μM; 24, 48 and 72 hours) treatment to HepG2 and Chang cells resulted in a dose and time dependent inhibition of cell growth as assessed by MTT and LDH assays. The data suggest that goniothalamin selectively inhibits HepG2 cells (IC50 of MTT= 4.6(±0.23) μM; IC50 of LDH= 5.20(±0.01) μM for 72 hours) with less inhibition of growth in Chang cells (IC50 of MTT= 35.0(±0.09) μM; IC50 of LDH= 32.5(± 0.04) μM for 72 hours. The cytotoxic activity of goniothalamin on HepG2 cells was confirmed by Trypan blue dye exclusion assay. Goniothalamin reduced the number of viable cells (non-stained) associated with an increase on the number of non-viable cells (stained) and the Viability Indexes were 52 ± 1.73% for HepG2 cells and 62 ± 4.36% for Chang cells at IC50 after 72 hours. Cells were exposed to goniothalamin at lowest concentration (2.3 μM), IC50 (of MTT results), and highest concentration (150 μM) for 24, 48, or 72 hours and then examined for effects on cell cycle (using the flow cytometry) or proliferation (using the BrdU ELISA assay). The cytotoxic activity of goniothalamin was related to the inhibition of DNA synthesis, as revealed by the reduction of BrdU incorporation. At 72 hours with the lowest goniothalamin concentration of 2.3 μM, the normal liver Chang cells retained 97.6% of control proliferation while the liver cancer HepG2 cells were reduced to 19.8% of control proliferation. Goniothalamin caused the accumulation of hypodiploid apoptotic cells in cell cycle analysis by flow cytometry. Goniothalamin arrested HepG2 and Chang cells in the G2/M phase with different degrees. Light microscopy examination of HepG2 and Chang cells exposed to different concentrations of goniothalamin up to 72 h demonstrated changes in cellular morphology; i.e. cell rounding followed by a loss of adherence with subsequent cell shrinkage and blebbing. In addition, the apoptotic cells were more abundant in goniothalamin-treated HepG2 cells (84 ± 4.58%) for 72 hours than in untreated cell (4 ± 2.65%) upon measurement by TUNEL staining. In view of the toxicity of goniothalamin, the kind of cell death, namely apoptosis or necrosis, was assessed. Therefore, staining with fluorescence labeled annexin V in combination with propidium iodide was performed on HepG2 and Chang cells exposed to goniothalamin. The laser scanning cytometry of propidium iodide and annexin V-stained cells indicated that the growth inhibiting effect of goniothalamin was consistent with a strong induction of apoptosis at late stage. This is because the cellular membrane integrity was lost, so the cells exhibited annexin V- and propidium iodidedouble positive up to 85.87 ± 0.78 and 57.69 ± 1.12 in HepG2 and Chang cells after 24 hours, respectively. In order to confirm apoptotic mechanism in the goniothalamintreated cells, caspase 3 activity upon the same treatment conditions was carried out. The results indicate that caspase 3 activity was significantly elevated early in IC50 treated Chang cells (574% of control) after 24 hours and late in IC50 treated cells after 72 hours in HepG2 cells (879% of control). Our findings suggest a potential mechanism for the strong growth inhibitory effect of goniothalamin on this HepG2 liver cancer cells. However, less sensitivity to normal liver Chang cell line was observed by this compound. An important feature of the cytotoxicity by goniothalamin is that it is mediated through apoptosis

A novel biocoagulant agent from mushroom chitosan as water and wastewater therapy

A new commercial cationic polyelectrolyte chitosan (CM), obtained from the waste of mushroom production, was examined using models of water and wastewater namely kaolin and palm oil mill effluent (pome). As it is biocompatible, widely available, and economically feasible, chitosan mushroom has high potential to be a suitable replacement for alum. Also, it can be a promising alternative to chitosan obtained traditionally from Crustaceans due to its higher zeta potential and homogeneity based on the raw material required for its production. A wide range of coagulant dose (5-60 mg l-1) and wastewater pH (2-12) were taken into account to find the optimal conditions of coagulation. The optimal doses are 10 and 20 mg l-1 at best pH (11 and 3) when treated with kaolin and palm oil mill effluent, respectively, while 1200 mg l-1 of alum was not enough to reach the efficiency of chitosan mushroom. On the other hand, the optimum dose of chitosan mushroom (20 mg l-1) at pH 3 of pome produced (75, 73, and 98%) removal of chemical oxygen demand (COD), biological oxygen demand (BOD), and total suspended solids (TSS), respectively. The significant potential of chitosan mushroom was proved by zeta potential measurement. Indeed, it possesses the highest zeta potential (+70 mV) as compared to the traditional chitosan produced from crustaceans. In short, chitosan mushroom as a biocoagulant is eco-friendly and it enhances water quality that meets the requirements of environmental conservatives

The effect of sulfate contents on the surface properties of iron–manganese doped sulfated zirconia catalysts

The iron–manganese doped sulfated zirconia catalysts were prepared via precipitation method; the sulfation was carried out by impregnation with different amounts of sulfate (4%, 10% and 16% SO4− 2 by weight) with the addition of Fe–Mn doped and calcined at 600 °C for 3 h. The prepared catalysts were characterized by TGA-DTA, XRD, BET, FT-IR, TEM, TPD-NH3 and XPS. XRD and BET results revealed that the addition of sulfate imparts special stabilization to the catalytically active tetragonal phase of zirconia. All the iron–manganese doped sulfated zirconia catalysts were found to have strong acid sites, high surface area and small crystallite size

Effect of recombinant human erythropoietin and doxorubicin in combination on the proliferation of MCF-7 and MDA-mb231 breast cancer cells

Patients with cancer often exhibit signs of anemia as the result of the disease. Thus, cancer chemotherapies often include erythropoietin (EPO) in the regime to improve the survival rate of these patients. The aim of the present study was to determine the effect of EPO on doxorubicin-treated breast cancer cells. The cytotoxicity of doxorubicin alone or in combination with EPO against the MCF-7 and MDA-MB 231 human breast cancer cells were determined using an MTT cell viability assay, neutral red (NR) uptake assay and lactate dehydrogenase (LDH) assay. The estimated half maximal inhibitory concentration values for doxorubicin and the combination of doxorubicin with EPO were between 0.140 and 0.260 µg/ml for all cells treated for 72 h. Treatment with doxorubicin in combination with EPO led to no notable difference in cytotoxicity, compared with treatment with doxorubicin alone. The antiproliferative effect of doxorubicin at a concentration of 1 µg/ml on the MDA MB 231 cells was demonstrated by the decrease in viable cells from 3.6x10(5) at 24 h to 2.1x10(5) at 72 h of treatment. In order to confirm apoptosis in the doxorubicin-treated cells, the activities of caspases-3/7 and 9 were determined using a TBE assay. The results indicated that the activities of caspases-3/7 and 9 were significantly elevated in the doxorubicin-treated MDA-MB-231 cells by 571 and 645%, respectively, and in the MCF 7 cells by 471 and 345%, respectively, compared with the control cells. EPO did not modify the effect of doxorubicin on these cell lines. The results of the present study suggested that EPO was safe for use in combination with doxorubicin in the treatment of patients with breast cancer and concurrent anemia

Characterization, drug release profile and cytotoxicity of dentatin-hydroxypropyl-β-cyclodextrin complex

This current work has been conducted mainly to increase solubility and drug release properties for high hydrophobic Dentatin (DEN) by incorporation it into Hydroxypropyl-β-Cyclodextrin (HPβCD) cavity. To confirm that inclusion be succeeded, the produced complex were installed onto different machines. The latter includes: Fourier transform infrared spectroscopy (FT-IR), X-ray diffractometry (XRD), differential scanning calorimetry (DSC), and field emission-scanning electron microscopy (FE-SEM). The hydrodynamic diameter and zeta potential of DEN-HPβCD complex were 2.025 ± 0.39 nm and −33.6 mV, respectively. Ultra-violet spectroscopy was employed to further confirmation of complexation process as well as to determine drug release profile. The result showed an initial burst release (19.9% within first two minutes) and then a continuous release for an extended period of 41 h (100%). The solubility of DEN was enhanced by >300 fold following complexation when a compared to DEN alone. Moreover, MTT finding showed that this complexation did not reduce cytotoxicity of DEN after applying on prostate cancer (LNCaP), human adenocarcinoma breast cancer (MDA-MB-231) and human gastric adenocarcinoma cell line (HDT). However, further investigations are required to validate efficacy of our produced inclusion using molecular analysis and in vivo studies

Cytotoxicity and physicochemical characterization of iron–manganese-doped sulfated zirconia nanoparticles

Iron–manganese-doped sulfated zirconia nanoparticles with both Lewis and Brønsted acidic sites were prepared by a hydrothermal impregnation method followed by calcination at 650°C for 5 hours, and their cytotoxicity properties against cancer cell lines were determined. The characterization was carried out using X-ray diffraction, thermogravimetric analysis, Fourier transform infrared spectroscopy, Brauner–Emmett–Teller (BET) surface area measurements, X-ray fluorescence, X-ray photoelectron spectroscopy, zeta size potential, and transmission electron microscopy (TEM). The cytotoxicity of iron–manganese-doped sulfated zirconia nanoparticles was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays against three human cancer cell lines (breast cancer MDA-MB231 cells, colon carcinoma HT29 cells, and hepatocellular carcinoma HepG2 cells) and two normal human cell lines (normal hepatocyte Chang cells and normal human umbilical vein endothelial cells [HUVECs]). The results suggest for the first time that iron–manganese-doped sulfated zirconia nanoparticles are cytotoxic to MDA-MB231 and HepG2 cancer cells but have less toxicity to HT29 and normal cells at concentrations from 7.8 µg/mL to 500 µg/mL. The morphology of the treated cells was also studied, and the results supported those from the cytotoxicity study in that the nanoparticle-treated HepG2 and MDA-MB231 cells had more dramatic changes in cell morphology than the HT29 cells. In this manner, this study provides the first evidence that iron–manganese-doped sulfated zirconia nanoparticles should be further studied for a wide range of cancer applications without detrimental effects on healthy cell functions

Physicochemical characterization and cancer cell antiproliferative effect of silver-doped magnesia nanoparticles

Silver-doped magnesia nanoparticles (Ag/MgO) were synthesized using the precipitation method and characterized by various techniques such as X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), thermal gravimetric analysis (TGA), Brunner-Emmett-Teller (BET) surface area measurements, and dispersive X-ray spectroscopy (EDX). The morphology of Ag/MgO nanoparticles was determined by transmission and scanning electron microscopy, which revealed cuboidal shaped nanoparticles with sizes ranging from 31 to 68 nm and an average size of 43.5 ± 10.6 nm. The anticancer effects of Ag/MgO nanoparticles were evaluated on human colorectal (HT29) and lung adenocarcinoma (A549) cell lines, and their caspase-3, -8, and -9 activities, as well as Bcl-2, Bax, p53, cytochrome C protein expressions were estimated. Ag/MgO nanoparticles showed selective toxicity towards HT29 and A549 cells while remaining relatively innocuous towards the normal human colorectal, CCD-18Co, and lung, MRC-5 cells. The IC50 values of Ag/MgO nanoparticles on the HT29 and A549 cells were found to be 90.2 ± 2.6 and 85.0 ± 3.5 μg/mL, respectively. The Ag/MgO nanoparticles upregulated caspase-3 and -9 activities, downregulated Bcl-2, upregulated Bax and p53 protein expressions in the cancer cells. The morphology of the Ag/MgO nanoparticle treated HT29 and A549 cells was typical of apoptosis, with cell detachment, shrinkage, and membrane blebbing. The results suggest that Ag/MgO nanoparticles induce apoptosis in cancer cells and exhibit potential as a promising anticancer agent.The publication of this article was funded by the Qatar National Library

Cytotoxicity of nickel zinc ferrite nanoparticles on cancer cells of epithelial origin

In this study, in vitro cytotoxicity of nickel zinc (NiZn) ferrite nanoparticles against human colon cancer HT29, breast cancer MCF7, and liver cancer HepG2 cells was examined. The morphology, homogeneity, and elemental composition of NiZn ferrite nanoparticles were investigated by scanning electron microscopy, transmission electron microscopy, and energy dispersive X-ray spectroscopy, respectively. The exposure of cancer cells to NiZn ferrite nano-particles (15.6-1,000 μg/mL; 72 hours) has resulted in a dose-dependent inhibition of cell growth determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The quantification of caspase-3 and -9 activities and DNA fragmentation to assess the cell death pathway of the treated cells showed that both were stimulated when exposed to NiZn ferrite nanoparticles. Light microscopy examination of the cells exposed to NiZn ferrite nanoparticles demonstrated significant changes in cellular morphology. The HepG2 cells were most prone to apoptosis among the three cells lines examined, as the result of treatment with NiZn nanoparticles. In conclusion, NiZn ferrite nanoparticles are suggested to have potential cytotoxicity against cancer cells

Curcumin gum Arabic nanoparticles demonstrate potent antioxidant and cytotoxic properties in human cancer cells

The main purpose of the study was to enhance the stability and therapeutic effects of Curcumin (Cur) through nanoformulation with gum Arabic (GA) as a coating agent through an efficient synthetic approach. The antioxidant properties of the developed nanoparticles (Cur/GANPs) were assessed through several in vitro assays, such as β-carotene bleaching activity, DPPH, and nitric oxide scavenging activities in addition to evaluating its inhibitory activity on angiotensin-converting enzyme (ACE). The cytotoxicity of Cur/GANPs was evaluated in vitro using different types of human cancer cells including breast cancer (MCF7, MDA-MB231), liver cancer (HepG2), and colon cancer (HT29) cells. The prepared particles displayed an elliptical shape with a size ranging between 20–260 nm and a potential difference of –15 mV. The Cur/GANPs exhibited significant antioxidant activity compared to free curcumin when using concentrations between 31.5 and 500 µg/mL. The Cur/GANPs also had inhibited the growth of all cancer cell lines in a proportional trend with concentrations used. Hence, the encapsulation with gum Arabic has augmented the antioxidant and anti-neoplastic effects of Curcumin. Therefore, Cur/GANPs may have effective therapeutic properties in diseases attributed to oxidative stress like cancer and hypertension

Preparation, characterization, in vitro drug release and anti-inflammatory of thymoquinone-loaded chitosan nanocomposite

In this study, we formulated Thymoquinone-loaded nanocomposites (TQ-NCs) using high-pressure homogenizer without sodium tripolyphosphate. The TQ-NCs were characterized and their anti-inflammatory determined by the response of the LPS-stimulated macrophage RAW 264.7 cells in the production of nitric oxide, prostaglandin E2, tumor necrosis factor-α, interleukin-6, and interleukin-1β. The physicochemical properties of TQ-NC were determined using different machines. TQ was fully incorporated in the highly thermal stable nanoparticles. The nanoparticles showed rapid release of TQ in the acidic medium of the gastric juice. In medium of pH 6.8, TQ-NC exhibited sustained release of TQ over a period of 100 h. The results suggest that TQ-NC nanoparticles have potential application as parenterally administered therapeutic compound. TQ-NC effectively reduce production of inflammatory cytokines by the LPS-stimulated RAW 264.7 cells, indicating that they have anti-inflammatory properties. In conclusion, TQ-NC nanoparticles have the characteristics of efficient carrier for TQ and an effective anti-inflammatory therapeutic compound.The publication of this article was funded by the Qatar National Library