MCF-7 Human Breast Carcinoma Cells Research Articles

Fatty Acids and Other Chemical Compositions of Some Seagrasses Collected from the Saudi Red Sea with Potential of Antioxidant and Anticancer Agents

The fatty acids (FAs), amino acids (AAs), phenolic and polysaccharides from Cymodocea serrulata, Halodule uninervis, Halophila stipulacea and Thalassia hemprichii were studied, their antioxidant and anticancer activities were also investigated. The results showed that T. hemprichii and C. serrulata were rich in gallic acid and protocatechuic acid with values of 45.26 and 25.26 mg/g, respectively. The polysaccharides of T. hemprichii and H. stipulacea were mainly composed of galactose (~ 21%), mannose (~ 14.5%) and glucose (~ 12.5%). The protein contents ranged from 3.06% (H. stipulacea) to 9.56% (T. hemprichii). Palmitic acid (C16:0) percentage was more than half of the total FAs content of H. uninervis and T. hemprichii, while linoleic acid (C18:2) was 35.7 and 28.6%, respectively, in both seagrasses. The highest steroid content was found in C. serrulata at 92.67%, and triterpenes lupeol (15.51%) and β-amyrin (16.59%) in H. stipulacea as the major oxygenated-unsaponifiable matters. T. hemprichii extract demonstrated the highest effective antioxidant activity with IC50 value of 1.2 µg/ml, while H. uninervis extract displayed potential cytotoxic effect against human SKOV-3 ovarian and MCF-7 breast carcinoma cells with IC50 values of 0.52 and 2.3 µg/ml, respectively.

Evaluation of Antiviral, Antibacterial and Antiproliferative Activities of the Endophytic Fungus Curvularia papendorfii, and Isolation of a New Polyhydroxyacid.

An endophytic fungus isolated from Vernonia amygdalina, a medicinal plant from Sudan, was taxonomically characterized as Curvularia papendorfii. Ethyl acetate crude extract of C. papendorfii revealed an important antiviral effect against two viral pathogens, the human coronavirus HCoV 229E and a norovirus surrogate, the feline coronavirus FCV F9. For the last one, 40% of the reduction of the virus-induced cytopathogenic effect at lower multiplicity of infection (MOI) 0.0001 was observed. Selective antibacterial activity was obtained against Staphylococcus sp. (312 µg/mL), and interesting antiproliferative activity with half maximal inhibitory concentration (IC50) value of 21.5 ± 5.9 µg/mL was observed against human breast carcinoma MCF7 cell line. Therefore, C. papendorfii crude extract was further investigated and fractionated. Twenty-two metabolites were identified by gas chromatography coupled to mass spectrometry (GC–MS), and two pure compounds, mannitol and a new polyhydroxyacid, called kheiric acid, were characterized. A combination of spectroscopic methods was used to elucidate the structure of the new aliphatic carboxylic acid: kheiric acid (3,7,11,15-tetrahydroxy-18-hydroxymethyl-14,16,20,22,24-pentamethyl-hexacosa-4E,8E,12E,16,18-pentaenoic acid). Kheiric acid showed an interesting result with a minimum inhibitory concentration (MIC) value of 62.5 µg/mL against meticillin-resistant Staphylococcus aureus (MRSA). Hence, endophytes associated with medicinal plants from Sudan merit more attention, as they could be a treasure of new bioactive compounds.

Abstract 2759: Immune cell killing assay for analyzing the effects of immunomodulating agents in vitro

Abstract Immuno-oncology has emerged as one of the most promising new approaches in cancer drug development. Greater cancer cell specificity of treatments can be achieved when the immune cells of a patient are used against tumors. Therefore, it is important to study the effects of immune cells on cancer cells using different models in vitro and in vivo. Our aim was to validate an in vitro T cell killing assay where activated peripheral blood mononuclear cells (PBMCs) target different cancer cells and increase their apoptosis. MCF-7 human breast carcinoma cells (ATCC) were used as target cells and were seeded on 96-well plates. 24 hours later, human PBMCs (Lonza) were added to the culture as effector cells. Simultaneously, the PBMCs were activated with 100 ng/ml anti-CD3 and 10 ng/ml IL2. Apoptosis was detected using IncuCyte Annexin V reagent, which was introduced to the cells at the same time with activators. Cell growth, proliferation and apoptosis were monitored with a live cell imager IncuCyte S3 (Sartorius) for three days. Cell viability was determined by CellTiter Glo (Promega) at the end of the study. The target-to-effector ratio of 1:5 and 1:10 both showed an increase in apoptosis of MCF-7 cells in the wells with activated PBMCs. This was detected with a confluence parameter (percentage) of IncuCyte S3. The T cell killing assay can be utilized for studying cancer therapeutics based on immune-oncology (e.g. pembrolizumab and atezolizumab) and to evaluate the potential of immune cells in killing cancer cells in the presence of drugs affecting the target-effector interaction. Citation Format: Jenni H. Mäki-Jouppila, Jussi M. Halleen, Katja M. Fagerlund. Immune cell killing assay for analyzing the effects of immunomodulating agents in vitro [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2759.

Nucleosomes effectively shield DNA from radiation damage in living cells.

Eukaryotic DNA is organized in nucleosomes, which package DNA and regulate its accessibility to transcription, replication, recombination and repair. Here, we show that in living cells nucleosomes protect DNA from high-energy radiation and reactive oxygen species. We combined sequence-based methods (ATAC-seq and BLISS) to determine the position of both nucleosomes and double strand breaks (DSBs) in the genome of nucleosome-rich malignant mesothelioma cells, and of the same cells partially depleted of nucleosomes. The results were replicated in the human MCF-7 breast carcinoma cell line. We found that, for each genomic sequence, the probability of DSB formation is directly proportional to the fraction of time it is nucleosome-free; DSBs accumulate distal from the nucleosome dyad axis. Nucleosome free regions and promoters of actively transcribed genes are more sensitive to DSB formation, and consequently to mutation. We argue that this may be true for a variety of chemical and physical DNA damaging agents.

Centaurea bruguierana inhibits cell proliferation, causes cell cycle arrest, and induces apoptosis in human MCF-7 breast carcinoma cells.

Centaurea bruguierana, of the Asteraceae family, has a long history of use in traditional medicines for the treatment of various ailments. However, the anticancer activity and underlying mechanisms have not yet been assessed. The C. bruguierana was extracted with methanol and fractionated into four different fractions. Different cancer cells and one non-cancerous were used to examine the cytotoxic effects of these fractions using MTT assay. The most potent fraction, C. bruguierana ethyl acetate fraction (CB EtOAc), was explored for its effects on cell cycle progression and apoptosis induction by Hoechst staining and annexin V-PI double staining in MCF-7 cells. The expression of apoptosis-related genes was quantified by RT-PCR. Of all fractions, CB EtOAc was found to have the strongest antiproliferative activity (IC50 = 10μg/mL) against MCF-7 cells. The antiproliferative activity of the CB EtOAc fraction against MCF-7 was correlated with arrested of cell cycle in the G1 phase, nuclear fragmentation, and the exposure of phosphatidylserine. The induction of apoptosis by CB EtOAc in MCF-7 cells was also associated with an increase in the Bax/Bcl-2 ratio and higher expression of caspases. Overall, our results demonstrated that CB EtOAc showed apoptosis-inducing effects, suggesting that C. bruguierana may be a promising source for a novel chemotherapeutic agents for the treatment of breast cancer.

Synthesis, cytotoxicity and in vitro antibacterial screening of novel hydrazones bearing thienopyridine moiety as potent COX-2 inhibitors

The starting precursors 2-carbohydrazides, bearing thienopyridine moiety, were prepared using 2-thioxopyridine-3-carbonitriles as key synthons. Next, 2-carbohydrazides were reacted with a variety of 4-substituted benzylidinemalononitriles or 4-substituted benzaldehydes to afford a new series of the target hydrazones incorporating thienopyridine moiety. The elemental analyses and spectral data were used to demonstrate the structures of new hydrazones series. The in vitro antibacterial activities of the target hydrazones were evaluated against different strains of Gram-positive and Gram-negative bacteria. In comparison with chloramphenicol as a reference drug, hydrazones 13b, 13c and 13d, linked to p-Cl, p-Br and p-Me moiety, respectively, exhibited the strongest activities against all tested bacteria with MIC values in the range of 6.2–12.5 μg/mL. In addition, several new hydrazones were tested as in vitro cytotoxic agents against each of human breast carcinoma MCF-7 cell line, colon cancer Caco2 cell line and liver hepatocellular carcinoma HEPG2 cell line. The hydrazones 13b, 13c and 13d demonstrated the best cytotoxicity against the tested eukaryotic cells. Furthermore, both experimental and docking studies could predict the promising inhibitory activities of hydrazones 13c and 13d against COX-2 enzyme with IC50 of 0.110 and 0.104 µM, respectively, when compared with Celecoxib (IC50 of 0.115 µM).

Redox-sensitive polyglutamic acid-platinum(IV) prodrug grafted nanoconjugates for efficient delivery of cisplatin into breast tumor

Targeting cisplatin to the sites of action and decreasing its side effects are still major challenges. Here, we introduced a polyglutamic acid-platinum(IV) prodrug nanoconjugates (γ-PGA-CA-Pt(IV)) constructed by polyglutamic acid and modified platinum(IV) prodrug to reserve the anti-tumor efficacy of cisplatin with decreased side effects. We describe the synthesis, physico-chemical characterization, and redox- and pH-sensitive releasing behavior of the nanoconjugate. In vitro studies revealed that, when incubated with glutathione in advance, the γ-PGA-CA-Pt(IV) nanoconjugate induced significant apoptosis in human breast carcinoma MCF-7 cells. From in vivo antitumor efficacy evaluation, the γ-PGA-CA-Pt(IV) nanoconjugate obviously improved the survival rate of tumor-bearing mice with inhibition of the tumor growth compared with cisplatin. Meanwhile, the nanoconjugates showed remarkable improved safety profile than the free cisplatin.

Synthesis of a novel fluorescent berberine derivative convenient for its subcellular localization study

Berberine is a naturally occurred isoquinoline alkaloid that shows great potential for developing anticancer drugs. However, the problem stays of poor understanding of the mechanisms of anticancer action of berberine. It depends on evaluation of berberine’s pharmacokinetics, namely monitoring of its uptake and distribution in cells, tissues and organs. In order to address these problems, we have designed and synthesized a novel berberine derivative BBR-BODIPY bearing a fluorescent tag that allows screening its interaction with the targeted cells. It was shown that the synthesized fluorescent derivative could penetrate into human breast carcinoma MCF7 cells, and then induced apoptosis detected by the Western Blot analysis as changed expression of apoptosis-related proteins, including Bax, Bcl2, and Cyto C released from mitochondria, Cleaved Caspase 9, Cleaved PARP, Pro-Caspase 3, and Cleaved Caspase 3. The results of MitoTracker analysis followed by the confocal microscopy of sub-cellular localization of BBR-BODIPY in the MCF7 cells demonstrated excellent cell-penetrating ability of this compound even at low concentrations, and mitochondria was the main site of its accumulation. Together with the results of Western Blot analysis, these data indicated that the mitochondria pathway might be involved in berberine-induced apoptosis.

Investigation of Silver Nitrate on Cytotoxicity and Apoptosis in MCF7 Human Breast Carcinoma Cells

Background: Metal compounds have been studied in vitro for many years and these compounds’s effects are shown on tumors. Anticancer potential of silver and silver metal compounds is investigated in these days. A study on the in vitro interactions of silver nitrate (AgNO3) with MCF7 human breast carcinoma cells was performed to detect cytotoxic effects which induce apoptotic pathways.Materials and Methods: The cytotoxicity of silver nitrate which administered on MCF7 cells was assessed by MTT assay. The apoptotic influences of silver nitrate (IC50: inhibition concentration) were determined using Annexin V-FITC/PI, JC-1, TUNEL paraffin embedded and confocal microscopy assays. Silver nitrate induced cytotoxicity and apoptosis in MCF7 cells. Results: In this work, we demonstrated that the inhibition of cell growth which is time and dose dependent in MCF7 cells for 24, 48 and 72 hours. The inhibition concentration of silver nitrate (IC50) was found as 10 µM in MCF7 cells for 72 hrs. The early/late apoptotic and necrotic changes which occured with silver nitrate (IC50: 10µM) administered, were analyzed in the MCF7 cells for 72 hrs. However, the reduced mitochondrial membrane activity (ΔΨmt) was observed by silver nitrate-treated (IC50: 10µM) in the MCF7 cells for 72 hrs. In addition to these findings, a variety of apoptotic structures were demonstrated on MCF7 cells for 72 hrs. Conclusions: The results suggest that silver nitrate could be attributed as chemotherapeutic agent for medical applications in breast cancer treatment.

Paratrimerin I, cytotoxic acridone alkaloid from the roots of Paramignya trimera

Bioactivity-guided fractionation of the CHCl3-soluble extract of the roots of Paramignya trimera was carried out to obtain a new acridone alkaloid, paratrimerin I. Its structure was elucidated based on NMR spectroscopic data interpretation. Paratrimerin I showed noteworthy cytotoxicity against the HepG2 human hepatocellular and MCF-7 human breast carcinoma cell lines, with the submicromolar IC50 values of 0.43 and 0.26 µM, respectively. The N-methyl, C-4 methoxy, and C-5 hydroxy groups in the acridone skeleton can be proposed as a structural feature for good cytotoxicity.

Ultra performance liquid chromatography-tandem mass spectrometeric analysis of ethyl acetate fraction from saudiLavandula coronopifoliaPoir and evaluation of its cytotoxic and antioxidant activities

Introduction:The ethyl acetate fraction of the Saudi Lavandula coronopifolia Poir has been previously reported to have hepatoprotective activity against ethanol-induced oxidative stress. The aim of the current study was to investigate the chemical composition, cytotoxic effect, and antioxidant activities of ethyl acetate fraction of the aerial parts of SaudiL. coronopifoliaPoir.Methods:Air dried aerial parts ofL. coronopifoliawere extracted using 90% ethyl alcohol. The dried extract was suspended in water, defatted with light petroleum and then fractionated with ethyl acetate. The ethyl acetate fraction was subjected to ultra performance liquid chromatography-tandem mass spectrometeric (UPLC-ESI/MS/MS) analysis in a negative ionization mode. The antioxidant activity of the fraction was determined using free radical 2,2-diphyenyl-picrylhydrazyl (DPPH) scavenging assay and its cytotoxic effect against HepG2 (human hepatocarcinoma) and MCF-7 (human breast carcinoma) cells were determined using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium (MTT) cell viability assay.Results:The major components of the ethyl acetate fraction included carvacrol-O-diglucoside, (34.98%) and trihydroxy ursolic acid (12.07%). Moreover, the DPPH radical scavenging activity of ethyl acetate fraction was measured. The ethyl acetate fraction revealed an antioxidant potential with EC5017.8 ± 1.3 µg/mL. Additionally, he ethyl acetate fraction showed cytotoxic activity against HepG-2 and MCF-7 cells with IC50=29.3 ± 0.9 µg/mL and 14.6 ± 0.3 µg/mL, respectively.Conclusion:The ethyl acetate fraction of the SaudiL. coronopifoliahas antioxidant activity and also cytotoxic activity against breast and liver cancer cells.

Structural characterization, antiproliferative and immunoregulatory activities of a polysaccharide from Boletus Leccinum rugosiceps

A homogeneous water-soluble flaky polysaccharide named LRP-1 (18.82 kDa) was extracted and purified from the rare edible and medicinal fungi Leccinum rugosiceps. The structure characterization of LRP-1 was analyzed by GPC, GC, GC–MS, FT-IR, SEM, 1D/2D NMR, and its antitumor and immunoregulatory activities were determined. The results show that LRP-1 is a new polysaccharide with a backbone which is mainly composed of →1-α-D-Fucp-3→, →1-α-D-Rhap-3→, →1-β-D-Glcp-6→, →1-β-D-Glcp-3→, →1-α-D-Glcp-4→, →1-β-D-Galp-6 → and →1-α-D-Manp-6→. Results of MTT assay show that LRP-1 inhibit the growth of human hepatoma cells HepG-2 and human breast carcinoma MCF-7 cells in vitro. LRP-1 could also induce the secretion of NO and immune factors (IL-6 and TNF-α) of RAW264.7. This study suggests that LRP-1 has potential to be used as nutraceutical food with antitumor and immunoregulatory activities.

Synthesis and cytotoxic evaluation of malachite green derived oleanolic and ursolic acid piperazineamides

The coupling of acetylated piperazinylamide spacered triterpenoic oleanolic acid and ursolic acid with meta or para substituted carboxylated malachite green analogs gave conjugates 10, 11, 15, and 16 that were cytotoxic for several human tumor cell lines. Especially, an oleanolic acid-derived compound 10 was cytotoxic for MCF-7 human breast carcinoma cells (EC50 = 0.7 μM). These derivatives represent first examples of triterpenoic acid derivatives holding a cationic scaffold derived from malachite green.

High Surface Reactivity and Biocompatibility of Y2O3 NPs in Human MCF-7 Epithelial and HT-1080 Fibro-Blast Cells

This study aimed to generate a comparative data on biological response of yttrium oxide nanoparticles (Y2O3 NPs) with the antioxidant CeO2 NPs and pro-oxidant ZnO NPs. Sizes of Y2O3 NPs were found to be in the range of 35±10 nm as measured by TEM and were larger from its hydrodynamic sizes in water (1004 ± 134 nm), PBS (3373 ± 249 nm), serum free culture media (1735 ± 305 nm) and complete culture media (542 ± 108 nm). Surface reactivity of Y2O3 NPs with bovine serum albumin (BSA) was found significantly higher than for CeO2 and ZnO NPs. The displacement studies clearly suggested that adsorption to either BSA, filtered serum or serum free media was quite stable, and was dependent on whichever component interacted first with the Y2O3 NPs. Enzyme mimetic activity, like that of CeO2 NPs, was not detected for the NPs of Y2O3 or ZnO. Cell viability measured by MTT and neutral red uptake (NRU) assays suggested Y2O3 NPs were not toxic in human breast carcinoma MCF-7 and fibroblast HT-1080 cells up to the concentration of 200 μg/mL for a 24 h treatment period. Oxidative stress markers suggested Y2O3 NPs to be tolerably non-oxidative and biocompatible. Moreover, mitochondrial potential determined by JC-1 as well as lysosomal activity determined by lysotracker (LTR) remained un-affected and intact due to Y2O3 and CeO2 NPs whereas, as expected, were significantly induced by ZnO NPs. Hoechst-PI dual staining clearly suggested apoptotic potential of only ZnO NPs. With high surface reactivity and biocompatibility, NPs of Y2O3 could be a promising agent in the field of nanomedicine.

DNA-dependent protein kinase regulates lysosomal AMP-dependent protein kinase activation and autophagy

ABSTRACT Macroautophagy/autophagy is a central component of the cytoprotective cellular stress response. To enlighten stress-induced autophagy signaling, we screened a human kinome siRNA library for regulators of autophagic flux in MCF7 human breast carcinoma cells and identified the catalytic subunit of DNA-dependent protein kinase PRKDC/DNA-PKcs as a positive regulator of basal and DNA damage-induced autophagy. Analysis of autophagy-regulating signaling cascades placed PRKDC upstream of the AMP-dependent protein kinase (AMPK) complex and ULK1 kinase. In normal culture conditions, PRKDC interacted with the AMPK complex and phosphorylated its nucleotide-sensing γ1 subunit PRKAG1/AMPKγ1 at Ser192 and Thr284, both events being significantly reduced upon the activation of the AMPK complex. Alanine substitutions of PRKDC phosphorylation sites in PRKAG1 reduced AMPK complex activation without affecting its nucleotide sensing capacity. Instead, the disturbance of PRKDC-mediated phosphorylation of PRKAG1 inhibited the lysosomal localization of the AMPK complex and its starvation-induced association with STK11 (serine/threonine kinase 11). Taken together, our data suggest that PRKDC-mediated phosphorylation of PRKAG1 primes AMPK complex to the lysosomal activation by STK11 in cancer cells thereby linking DNA damage response to autophagy and cellular metabolism. Abbreviations: AXIN1: axin 1; 3-MA: 3-methyladenine; 5-FU: 5-fluorouracil; AA mutant: double alanine mutant (S192A, T284A) of PRKAG1; ACACA: acetyl-CoA carboxylase alpha; AICAR: 5-Aminoimidazole-4-carboxamide ribonucleotide; AMPK: AMP-activated protein kinase; ATG: autophagy-related; ATM: ataxia telangiectasia mutated; ATR: ATM serine/threonine kinase; AV: autophagic vacuole; AVd: degradative autophagic vacuole; AVi: initial autophagic vacuole; BECN1: beclin 1; BSA: bovine serum albumin; CBS: cystathionine beta-synthase; CDK7: cyclin dependent kinase 7; CDKN1A: cyclin dependent kinase inhibitor 1A; EGFP: enhanced green fluorescent protein; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GST: glutathione S transferase; H2AX/H2AFX: H2A.X variant histone; HBSS: Hanks balanced salt solution; IP: immunopurification; IR: ionizing radiation; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MAP3K9: mitogen-activated protein kinase kinase kinase 9; mRFP: monomeric red fluorescent protein; mCh: mCherry; MCM7: minichromosome maintenance complex component 7; MTORC1: mechanistic target of rapamycin kinase complex 1; NHEJ: non-homologous end joining; NRBP2: nuclear receptor binding protein 2; NTC: non-targeting control; NUAK1: NUAK family kinase 1; PBS: phosphate-buffered saline; PIK3AP1: phosphoinositide-3-kinase adaptor protein 1; PIK3CA: phosphatidylinositol-4,5-biphosphate 3-kinase catalytic subunit alpha; PIKK: phosphatidylinositol 3-kinase-related kinase; PRKAA: protein kinase AMP-activated catalytic subunit alpha; PRKAB: protein kinase AMP-activated non-catalytic subunit beta; PRKAG: protein kinase AMP-activated non-catalytic subunit gamma; PRKDC: protein kinase, DNA-activated, catalytic subunit; RLuc: Renilla luciferase; RPS6KB1: ribosomal protein S6 kinase B1; SQSTM1: sequestosome 1; STK11/LKB1: serine/threonine kinase 11; TP53: tumor protein p53; TSKS: testis specific serine kinase substrate; ULK1: unc-51 like autophagy activating kinase 1; WIPI2: WD repeat domain, phosphoinositide interacting 2; WT: wild type.

Attenuated anti-tumor activity of NK-92 cells by invasive human breast carcinoma MDA-MB-231 cells

Natural killer (NK) cells are typical innate lymphocytes that directly kill cancer cells. However, the anti-tumor function of NK cells can be modulated by various cancers in a tumor microenvironment. We investigated whether the functional characteristics of NK cells are affected by coculture with human breast carcinoma MCF-7 or MDA-MB231 cells and further examined the underlying molecular mechanisms that are associated with attenuated anti-tumor activity of NK cells. NK-92 cells were cocultured with MCF-7 or MDA-MB-231 cells at a ratio of 1:2 (target:effector) for 6 h. The cytotoxic effect of NK-92 cells was measured using CytoTox96 assay. The frequency of CD56, NKp30+, NKp44+, NKG2D+, and NKG2A+ in NK-92 cells was investigated using individual fluorescent antibodies and flow cytometry. In addition, the production of cytokines was measured using Human Cytokine Array C1 kit. Proteins in cancer cell lysates were quantified and the expressions of PI3K, pAkt, Stat3, and pStat3 were examined by western blot analysis. The cytotoxicity of NK-92 cells is greater against MCF-7 than against MDA-MB-231 cells and cocultures with MCF-7 or MDA-MB-231 cells increased the frequency of CD56dim population in NK-92 cells as well as IFN-γ production. The frequency of the NKp30+ or NKG2D+ population in NK-92 cells was significantly decreased in coculture with MCF-7 or MDA-MB-231 cells. However, the frequency of NKG2A+ expression in NK-92 cells was significantly increased in coculture with MCF-7 or MDA-MB-231 cells. The secretions of IL-6, IL-8, monocyte chemoattractant protein-1, and granulocyte–macrophage colony-stimulating factor were significantly higher in supernatant from coculture of NK-92 and MDA-MB-231 cells than in supernatants from NK-92 cell single culture or coculture of NK-92 and MCF-7 cells, which positively correlates with the enhanced expression of pStat3 in MDA-MB-231 cells cocultured with NK-92 cells. These findings demonstrate that the anti-tumor function of NK-92 cells is significantly suppressed by an invasive human breast carcinoma, MDA-MB-231, and the impairment of NK-92 cell function is associated with the upregulation of the IL-6/Stat3 signaling pathway.

IN VITRO CYTOTOXIC AND APOPTOTIC EFFECT OF CONCANAVALINS FROM CANAVALIA spp. ON BREAST CARCINOMA CELLS

Concanavalins from Canavalia spp. possess diverse bioactivities and exhibit potent anticancer effects. In this study we assessed the cytotoxic activity of concanavalins from Canavalia spp. on human breast carcinoma MCF-7 cells. The lectins concanavalin A (ConA), concanavalin C (ConC), concanavalin G (ConG) and concanavalin M (ConM) were isolated from seeds of C. ensiformis, C. cathartica, C. gladiata and C. rosea respectively. ConAand ConG demonstrated strong inhibitory effect on MCF-7 cell proliferation with IC values 81.66 µg/mLand 81.09 µg/mLrespectively, with no 50 significant effect in normal kidney HEK293T cells. MCF-7 cells treated with ConA and ConG showed apoptotic morphological changes and accumulation of sub-G /G proportion, suggesting stimulation of apoptotic pathway. These 0 1 results uphold the anticancer potential of concanavalins.

Evaluation of the time-dependent antiproliferative activity and liver microsome stability of 3 phenyl 4-(2-oxo-3-alkylimidazolidin-1-yl)benzenesulfonates as promising CYP1A1-dependent antimicrotubule prodrugs

In this study, the antiproliferative activity of 3 phenyl 4-(2-oxo-3-alkylimidazolidin-1-yl)benzenesulfonates (PAIB-SOs) was assessed in a time-dependent manner together with their hepatic stability and metabolism using human, mouse and rat liver microsomes. CEU-818, -820 and -913 were selected as promising hit compounds. Their antiproliferative activity on human breast carcinoma MCF-7 cells was evaluated using escalating concentrations of drugs at 24, 36 and 48h and the sulforhodamine B assay. Their hepatic stability was evaluated by HPLC-UV of extracts obtained from human, mouse and rat liver microsomes. The antiproliferative activity of PAIB-SOs is concentration and time-dependent and requires between 24 and 36h of contact with MCF-7 cells to detect a significant antiproliferative activity. PAIB-SOs stability in microsomes usually decreases following this order: human ≈ (rat>mouse). The CEU-913 exhibits the longest half-life in rat and human liver microsomes while the CEU-820 exhibits the longest half-life in mouse liver microsomes. Our in vitro results suggest that PAIB-SOs should have a minimum contact time of 24h with the tumour to trigger significant antitumoural activity. The activity of mouse liver microsomes towards PAIB-SOs is higher than rat microsomes and tends to be higher than human liver microsomes.

Synthesis, characterization and in vitro evaluation of anticancer activity of a new water-soluble thiosemicarbazone ligand and its complexes

A new water-soluble thiosemicarbazone ligand, 4-hydroxyl-3-({[(methylamino)carbonothioyl] hydrazono}methyl)benzoic acid (H3LCOOH) and its complexes with Cu(II), Ni(II), Zn(II), Fe(III) and Mn(III) were synthesized and characterized. These compounds were characterized by conductivity, magnetic susceptibility, elemental analysis, FT-IR, 1H NMR, 13C NMR, and UV–Vis spectroscopy. All the compounds have sufficient solubility and stability in water as a safe, green, and biologic solvent. The compounds were evaluated for their anticancer activities against the human chronic myelogenous K562 leukemia and human breast carcinoma MCF-7 cell lines. All the complexes significantly showed more anticancer activity than the ligand alone and the Cu(II) complex showed considerable cytotoxicity against the tested cell lines. The mechanism of action of the most potent complex, Cu(HLCOO) against K562 cell line was further explored by fluorescence microscopy, flow cytometry, cell cycle and reactive oxygen species (ROS) assays. Hoechst staining and flow cytometry indicated the apoptosis induction ability of Cu(HLCOO) on K562 cells. Mechanistic studies indicated that Cu(HLCOO) complex caused cell cycle arrest at G0/G1 phase and this complex was able to inhibit the cell cycle in a time-dependent manner. Cu(HLCOO) can also increase the reactive oxygen species levels in the K562 cells. The obtained results suggest that the studied compounds can be helpful candidates for further evaluations as chemotherapeutic agents in the treatment of cancer.

Synthesis of novel metal (II) complexes tailored from 9-oxo-9H-fluorene-1-carboxylic acid via green protocol: DNA cleavage and anticancer studies

Despite being a polycyclic aromatic compound, fluorenone is not classified as carcinogenic. Some of the derivatives of fluorenone and metal complexes embedded with fluorenone based ligands exhibited challenging biological activities, including anticancer activity. The current investigation hence focuses on synthesis, characterization and anticancer activities of fluorenone based ligands and their coordinate compounds of some first row transition metals. The stoichiometry of all the synthesized metal complexes is found to be 1: 2 (metal: ligand) with the general formula [M(L)2], L = singly deprotonated ligand, whereas the geometry of all the metal complexes is found to be octahedral. The synthesized ligands and their respective metal (II) complexes were found to cleave the pBR322 DNA, during gel electrophoresis studies. The inhibition of cancer cell growth has been confirmed by fluorescence imaging techniques using DAPI as staining dye. The IC50 values of ligands and their metal (II) complexes suggest that synthesized compounds are more active towards A549 (human lung adenocarcinoma) cell line when compared with the standard drug Paclitaxel. In case of MCF-7 (human breast carcinoma) cell line, compound Cu(L2)2 is found to be more active than the standard drug Paclitaxel.