Ag-doped ZnO nanoparticles synthesized through green method using Artemisia turcomanica extract induce cytotoxicity and apoptotic activities against AGS cancer cells: an in vitro study

In recent years, probiotics and their derivatives have been recognized as important therapeutic agents in the fight against cancer. Therefore, this study aimed to investigate the anticancer effects of membrane vesicles (MVs) from Lentilactobacillus buchneri strain HBUM07105 probiotic isolated from conventional and unprocessed yogurt in Arak province, Iran, against gastric and colon cancer cell lines. The MVs were prepared from the cell-free supernatant (CFS) of L. buchneri and characterized using field-emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM) and SPS-PAGE techniques. The anticancer activity of MVs was evaluated using MTT, flow cytometry, qRT-PCR techniques, and a scratch assay. The study investigated the anti-adenocarcinoma effect of MVs isolated from L. buchneri on a human gastric adenocarcinoma cell line (AGS) and a human colorectal adenocarcinoma cell line (HT-29) at 24, 48, and 72-h time intervals. The results demonstrated that all prepared concentrations (12.5, 25, 50, 100, and 200 µg/mL) of MVs reduced the viability of both types of human adenocarcinoma cells after 24, 48, and 72 h of treatment. The analysis of the apoptosis results revealed that the percentage of AGS and HT-29 cancer cells in the early and late stages of apoptosis was significantly higher after 24, 48, and 72 h of treatment compared to the untreated cancer cells. After treating both AGS and HT-29 cells with the MVs, the cells were arrested in the G0/G1 phase. These microvesicles demonstrate apoptotic activity by increasing the expression of pro-apoptotic genes (BAX, CASP3, and CASP9). According to the scratch test, MVs can significantly decrease the migration of HT-29 and AGS cancer cells after 24, 48, and 72 h of incubation compared to the control groups. The MVs of L. buchneri can also be considered a potential option for inhibiting cancer cell activities.

Objective: To investigate the cytotoxic, apoptotic and inhibitory activities on cell migration and invasion of plumbagin in the human cholangiocarcinoma (CCA) cell line (CL-6) in comparison with human embryonic fibroblast cell line (OUMS). Methods: Cytotoxicity activity was evaluated using MTT assay. Inhibitory effect on cell migration and invasion were investigated using label-free real-time cell analysis and QCM ECMatrix cell invasion chamber, respectively. Apoptotic activity was evaluated using flow cytometry and CellEvent™ Caspase 3/7 assay. Results: Based on results of the cytotoxicity test in CL-6 cells, 50% inhibitory concentration (IC50, Mean±SD) values of plumbagin and the standard drug 5-fluorouracil were (24.00±3.33) and (1 036.00±137.77) μmol/L, respectively. The corresponding values for OUMS cells were (57.00±5.23) and (2 147.00±209.98) μmol/L, respectively. The selectivity index was 2.28. The inhibitory activities of plumbagin on cell migration and invasion were potent and concentration-dependent with IC50 of 25.0 μmol/L and complete inhibition at 25.0 μmol/L. Flow cytometry analysis showed that plumbagin at 12.5 μmol/L (half IC50) induced CL-6 cell apoptosis (43.24% of control) through stimulation of caspase 3/7 activities. Complete cell apoptosis was observed at 12.5 μmol/L. Conclusions: The cytotoxic activity and inhibition of migration and invasion including apoptosis induction in the human CCA cell line (CL-6) suggest that plumbagin could be a promising candidate for CCA chemotherapeutics. However, its relatively low selective cytotoxic effect on CCA cells is a major concern.

Gastric cancer (GC) is one of the most prevalent cancer types worldwide and one of the leading causes of cancer-related deaths. Gastric cancer is an aggressive and heterogeneous disease with a poorly understood carcinogenesis at the molecular level. Therefore, the research for effective drug therapy strategies plays a significant role in treating the disease. One of these effective treatment strategies is herbal-based therapeutics, which have low side effects and contain many biologically active compounds. In this study, the effect of U.dioica, S. marianum and C. scolymus herbal extracts on cell viability in L929, AGS and SH-SY5Y cell lines was analyzed by XTT test to evaluate the anticancer activities. Antibacterial and antioxidant activities of extracts were determined by the agar well diffusion test and CUPRAC method, respectively. We found that U. dioica and S. marianum extracts showed no significant effect on the viability of AGS and SH-SY5Y cancer cells. C. scolymus extract demonstrated strong anticancer activity on AGS cancer cells at all concentrations but had no effect on SH-SY5Y cells. U. dioica and C. scolymus exhibited antibacterial activity against S. aureus and B. cereus, respectively. No antibacterial activity was found in S. marianum extract. U. dioica and S. marianum extracts have shown strong antioxidant activity in CUPRAC assay. In conclusion, the obtained results revealed the antibacterial and anticancer therapeutic potential of C. scolymus extract known as artichoke in gastric cancer cells. However, more research is required to better explain the therapeutic properties of these extracts.

BackgroundCephalotaxus spp. are known to possess various therapeutic potentials. Cephalotaxus griffithii, however, has not been evaluated for its biological potential. The reason may be the remoteness and inaccessibility of the habitat where it is distributed. The main aim of this study was to: (1) evaluate multiple biological potentials of stem bark of C. griffithii, and (2) identify solvent extract of stem bark of C. griffithii to find the one with the highest specific biological activity.MethodsDried powder of stem bark of C. griffithii was exhaustively extracted serially by soaking in petroleum ether, acetone and methanol to fractionate the chemical constituents into individual fractions or extracts. The extracts were tested for total phenolic and flavonoid content, antioxidant (DPPH radical scavenging, superoxide radical scavenging, and reducing power models), antibacterial (disc diffusion assay on six bacterial strains), cytotoxic (MTT assay on HeLa cells), and apoptotic activity (fluorescence microscopy, DNA fragmentation assay, and flow cytometry on HeLa cells).ResultsAmong the three extracts of stem bark of C. griffithii, the acetone extract contained the highest amount of total phenolics and flavonoids and showed maximum antioxidant, antibacterial, cytotoxic (IC50 of 35.5 ± 0.6 μg/ml; P < 0.05), and apoptotic (46.3 ± 3.6% sub-G0/G1 population; P < 0.05) activity, followed by the methanol and petroleum ether extracts. However, there was no significant difference observed in IC50 values (DPPH scavenging assay) of the acetone and methanol extracts and the positive control (ascorbic acid). In contrast, superoxide radical scavenging assay-based antioxidant activity (IC50) of the acetone and methanol extracts was significantly lower than the positive control (P < 0.05). Correlation analysis suggested that phenolic and flavonoid content present in stem bark of C. griffithii extracts was responsible for the high antioxidant, cytotoxic, and apoptotic activity (P < 0.05).ConclusionsStem bark of C. griffithii has multiple biological effects. These results call for further chemical characterization of acetone extract of stem bark of C. griffithii for specific bioactivity.

Comparison of different properties of zinc oxide nanoparticles synthesized by the green (using Juglans regia L. leaf extract) and chemical methods

This review describes the state of the art regarding the prebiotic role of chitin and the interactions of chitin and chitosan with cancer cells. Chitin is the second most abundant polysaccharide in nature and a constitutive component of crustacean shells and the exoskeleton of insects. Chitosan is the deacetylated form of chitin, which is obtained by chemical processing or the enzymatic activity of deacetylases found in microorganisms and insects. Edible insects have recently been introduced in Western countries, thus raising concerns regarding food safety and due to their chitin content and the release of chitosan during the digestive process. The roles of insect chitin and chitosan in the gastrointestinal tract, microbiome modulation, and cancer have been widely investigated. Several in vitro and in vivo studies have shown the possible microbiota modulation of chitin and its relevant communication with the immune system, thus confirming its prebiotic activity. No evidence has been provided on the cancerogenic activity of chitin; however, studies have suggested that chitin has a cytotoxic effect on cancer cell lines. Chitosan has been confirmed to exhibit apoptotic and cytotoxic activities on cancer cells in several in vitro studies on cancer cell lines and in vivo models. In conclusion, the literature does not show a direct connection between the presence of chitin or chitosan and the onset of cancer. However, cytotoxic and apoptotic activities in relation to cancerous lines have been demonstrated.

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In vitro cytotoxic and toxicological activities of ethanolic extract of Kaempferia galanga Linn. and its active component, ethyl-p-methoxycinnamate, against cholangiocarcinoma

There are thousands of herbs in Thailand used in traditional medicine that are claimed to possess anticancer activity, but most of them have no scientific evidences to support their alleged medicinal quality. One among them is Stephania venosa (Bl.) spreng or S.venosa. The tuber of S.venosa has been used in many Thai traditional medicines as remedy of various diseases including cancer. In this study, we aim to apply cell culture technique and design a scientific model for testing and proving the claimed anticancer activity of these Thai herbs. Lymphocytes obtained from healthy blood donors were cultured in RPMI medium with the density of 4x10[superscript 5] cells/ml and used in various tests throughout the study. The inhibitory concentration at 50% (IC[subscript 50]) of water extract of S.venosa was determined by exposing 4x10[superscript 5] lymphocytes to various concentrations of S.venosa for 48 hours, and trypan blue dye exclusion was the technique applied to detect the cytotoxic activity. Detection of pH change and the cytotoxic activity of aliquots of S.venosa were studied for a period of 12 weeks to monitor the stability of the water extract of the herb. Besides the study of cytotoxic activity, IC[subscript 50] of water extract at 300 microgram/ml was selected for the test of apoptotic activity, both in normal human lymphocytes and lymphocytes obtained from cervical cancer patients. Both sources of lymphocytes were exposed to various treatments including: 100 microgram/ml S.venosa, 300 microgramm/ml S.venosa, 0.5 Gy [superscript 60]Co irradiation and a combination of 300 microgram/ml S.venosa plus 0.5 Gy [superscript 60]Co irradiation. The apoptotic activity was detected at 48 hour after exposure by in situ terminal deoxynucleotidyl transferase assay (TdT assay). Furthermore, inhibitory effect of various concentrations of water extract of S.venosa was tested on PHA stimulated normal human lymphocytes. Results from all studies indicated that, water extract of S.venosa tuber possessed cytotoxicity and apoptotic activities against both normal lymphocytes and lymphocytes obtained from cervical cancer patients. A combination of 0.5 Gy [superscript 60]Co irradiation with 300 microgram/ml S.venosa exhibited additive effect when detected by TdT assay. Besides, water extract of S.venosa exhibited definite inhibitory activity on PHA stimulated normal human lymphocytes at IC[subscript 50] of 40 microgram/ml. With the above data, it can then be primarily concluded that water extract of S.venosa should be further investigated and developed in line of other anticancer herbal drugs.

Blumea lacera (B. lacera) (Asteraceae) is a well-known Bangladeshi medicinal plant. This study aimed to identify and characterize constituents associated with the significant cytotoxic activity of this plant that we reported previously. Here, we describe the isolation and characterization of a new steroidal glycoalkaloid (SGA) 1, the evaluation of its cytotoxic activity, apoptotic potential, and effect on cell cycle in comparison to analogous steroidal glycoalkaloids (SGAs). SGA 1 was isolated using C18 SPE and HPLC, and subsequently structurally characterized using 1D and 2D NMR, MS and other spectroscopic methods, along with a comparative inspection of the literature. Cytotoxic activity of 1 and seven SGA analogues and steroidal alkaloids (SAs), (β-solamarine, α-solanine, β-solamargine, α-solasonine, khasianine, solasodine, tomatidine HCl) were evaluated for their cytotoxicity against two healthy (NIH3T3 and VERO) and four human cancer (AGS, HT-29, MCF-7 and MDA-MB-231) cell lines using the MTT assay. Cytotoxic SGAs were further evaluated for apoptosis-inducing potential and cell cycle arresting ability against breast cancer cells (MCF-7) using the FITC Annexin V and propidium iodide (PI) assay. Bioactivity guided fractionation of the methanol extract of B. lacera led to isolation of compound 1: (25R)-3β-{O-β-D-glucopyranosyl-(1 → 4)-O-α-L-rhamnopyranosyl-(1 → 4)-[O-α-L-rhamnopyranosyl-(1 → 2)]-α-L-rhamnopyranosyl}-22αN-spirosol-5-ene. SGA 1 was the most cytotoxic compound against a number of human cancer cell lines with an IC50 of 2.62 µM against MCF-7 cells. It displayed the highest apoptotic potential (32% AV+/PI-) on MCF-7 cells compared to other cytotoxic SGA analogues and a slight, but significant cell cycle arresting effect. A new SGA 1 was isolated from B. lacera and its cytotoxic activity, as well as that of other SAGs, was evaluated. SAR investigations on SGA 1, in relation to SGA analogues, show that the number and nature of sugar moieties along with the linkages of the sugar to the aglycone are crucial for cytotoxic and apoptotic activity. This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.

Although phthalocyanines are usually used a photosensitizers for photodynamic therapy, these works focus on the directly cytotoxic effect of a new water-soluble magnesium phthalocyanine. The new water-soluble magnesium phthalocyanine 2 was synthesized, characterized and investigated for cytotoxic and apoptotic activities. The cytotoxic activities of the compound 2 were determined by using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell viability assay on human breast cancer cells (MDA-MB-231, MCF-7), human prostate cancer cells (PC-3), and human healthy lung fibroblast cells (WI-38). The cells were plated and treated 1 to 20 [Formula: see text]M of different concentrations of the compound. MTT assay results indicated that the compound 2 has concentration and time-dependent cytotoxic activities against cancer cells. We also observed that the compound displayed lower toxicity against WI-38 healthy cells than cancer cells at 48 and 72 h. The compound showed a significant cytotoxic activity difference between breast cancer cells and WI-38 healthy cells at 48 and 72 h. Selectivity index of the compound 2 against MCF-7 for 72 h was calculated ¿ 15.62. We also studied the apoptotic and necrotic effect of compound 2using TUNEL and PI staining, respectively. It was found that the synthesized compound 2 increased apoptotic and necrotic cells.

BackgroundPyocyanin, a specific extracellular secondary metabolite pigment produced by Pseudomonas aeruginosa, exhibits redox activity and has toxic effects on mammalian cells, making it a new and potent alternative for treating cancer. Breast cancer (BC) treatment is now defied by acquired and de novo resistance to chemotherapy, radiation, or targeted therapies. Therefore, the anticancer activity of purified and characterized pyocyanin was examined against BC in our study.ResultsThe maximum production of pyocyanin (53 µg/ml) was achieved by incubation of the highest pyocyanin-producing P. aeruginosa strain (P32) in pH-adjusted peptone water supplemented with 3% cetrimide under shaking conditions at 37 °C for 3 days. The high purity of the extracted pyocyanin was proven by HPLC against standard pyocyanin. The stability of pyocyanin was affected by the solvent in which it was stored. Therefore, the purified pyocyanin extract was lyophilized to increase its shelf-life up to one year. Using the MTT assay, we reported, for the first time, the cytotoxic effect of pyocyanin against human breast adenocarcinoma (MCF-7) with IC50 = 15 μg/ml while it recorded a safe concentration against human peripheral blood mononuclear cells (PBMCs). The anticancer potential of pyocyanin against MCF-7 was associated with its apoptotic and necrotic activities which were confirmed qualitatively and quantitively using confocal laser scanning microscopy, inverted microscopy, and flow cytometry. Caspase-3 measurements, using real-time PCR and western blot, revealed that pyocyanin exerted its apoptotic activity against MCF-7 through caspase-3 activation.ConclusionOur work demonstrated that pyocyanin may be an ideal anticancer candidate, specific to cancer cells, for treating MCF-7 by its necrotic and caspase-3-dependent apoptotic activities.

This study aimed to investigate the cytotoxic effects of 2-amino thiazole compound (3) on MCF-7 breast and AGS gastric cancer cells. We examined the cytotoxic effects of various concentrations (10-25-50-100 μg/mL) of 2-aminothiazol compound (3) on MCF-7 breast cancer and AGS gastric cells at 24, 48 and 72 hours using MTT assay. MTT assays demonstrate that the 2-amino thiazole (3) compound has a time- and dose-dependent inhibitory effect on the proliferation of MCF-7 and AGS cancer cells. Analysis of the data obtained from MTT assay showed that IC50 values for thiazole were 80.13, 71.03 and 59.24 µg/ml and 75.03, 38.12 and 28.01 µg/ml for 24, 48 and 72 hours on MCF-7 and AGS cells, respectively. The 100 μg/mL dose was demonstrated to be most effective on both cancer cells. Our results suggest that the 2-amino-4-phenylthiazole (3) compound has a dose-dependent effect on cytotoxicity in MCF7 breast and AGS gastric cancer cells.

Background: The orchid plants of Dendrobium, belong to the largest family of flowering plants orchidaceae. The genus Dendrobium reported to possess various therapeutic activities including anticancer activity. In the present scenario, cancer is one of the most leading causes of death worldwide. Therefore, the main objective of the present study was to demonstrate the cytotoxic and apoptotic inducing activity of Dendrobium crepidatum and Dendrobium chrysanthum ethanolic extracts in T-cell lymphoma. Methods: MTT assay was done to evaluate the cytotoxic effect of the extracts on lymphoma cells. The reactive oxygen species (ROS) generation by the ethanolic extracts was also examined by DCFH-DA. Fluorescence microscopy studies and DNA fragmentation assay were done to investigate the apoptotic cell death. Flow cytometry was done to check the effect of ethanolic extracts on cell cycle phase distribution. Results: The results revealed that both the extracts led to dose dependent cytotoxic effect in T-cell lymphoma along with increased generation of intracellular reactive oxygen species in a dose and time dependent manner. The fluorescent microscopic studies showed that the extracts induced cellular shrinkage, chromatin condensation and nuclear fragmentation, the major hallmarks of apoptosis, in dose dependent manner. The presence of DNA ladder further confirms the process of nuclear fragmentation, and the cell cycle analysis showed significant delay at G2/M phase of the cell cycle with both the extracts. Conclusion: The whole study suggests that Dendrobium crepidatum and Dendrobium chrysanthum ethanolic extracts induced substantial cytotoxic and apoptotic activity in T-cell lymphoma. Therefore, further studies are needed to establish the detailed mechanism of anticancer activity of both the ethanolic extracts.

In vitro anticancer activity of fucoidan extracted from Sargassum cinereum against Caco-2 cells