Induces Apoptosis Cell Death Research Articles

Paclitaxel-betulinic acid hybrid nanosuspensions for enhanced anti-breast cancer activity

Paclitaxel-betulinic acid hybrid nanosuspensions (PTX-BA-NP) with increased anti-breast cancer activity were developed. The resultant nanosuspensions (NP) had a mean particle size of 282.54 ± 5.4 nm, a polydispersity index of approximately 0.242 ± 0.02, a zeta potential of -19.7 ± 0.19 mV and a redispersibility index of 103.3 ± 0.01%. The cumulative dissolution percentage of PTX coarse powder and PTX-BA-NP dried powder at 60 min were 15.4% and 90.8%, respectively. MCF-7 cell-based testing showed that treatment with PTX-BA-NP led to more PTX-BA-NP accumulation in the cytoplasm of breast cancer cells, less cell cycle arrest in the G2-M phase, more cell cycle arrest in the G0-G1 phase, more apoptosis-induced cell death and stronger inhibition of cell migration than paclitaxel nanosuspensions (PTX-NP). Biodistribution studies showed that tumor accumulation levels at 12 h in the PTX-BA-NP group were approximately 2.67- and 2.33-fold higher than the levels in the Taxol® and PTX-NP groups, respectively. In vivo antitumor efficacy demonstrated that PTX-BA-NP exerted the strongest tumor inhibition among the four groups, with a tumor inhibition rate of 47.79 ± 2.28%, followed by PTX-NP (35.05 ± 5.55%), Taxol® (22.67 ± 6.01%) and betulinic acid nanosuspension (BANP) (14.38 ± 6.02%). These findings indicate that PTX-BA-NP holds great promise for breast cancer therapy.

132 - Antioxidant and anticancer activities from Stryphnodendron adstringens

Stryphnodendron adstringens is a tree species native to the Brazilian Cerrado commonly known as barbatimao or casca-da-mocidade. In traditional medicine, the stem bark of this plant are used in the treatment of several diseases. Scientifically, its antifungal, wound healing, anti-inflammatory and antidiabetic properties have already been shown. The objective of this study was assess their antioxidant activity, and anticancer effects, and chemical composition of the aqueous extract from stem bark of Stryphnodendron adstringens (SBSA). The antioxidant activity of the SBSA was assessed using two free radical scavenging methods, the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays, oxidative hemolysis inhibition,and2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced lipid peroxidation inhibition in human erythrocytes. B16F10Nex-2 melanoma cells were used to assess the anticancer activity as well as the underlying mechanisms of SBSA-induced cell death. Phenolic compounds and flavonoids present in the SBSA were quantified using two colorimetric methods involving the Folin-Ciocalteu reagent and aluminum chloride complexation reaction, respectively. The concentration of phenolic compounds was 195.16 ± 0.94 mg GAE/g SBSA, and the concentration of flavonoids was 2.87 ± 0.08 mg QE/g. The SBSA showed the antioxidant activity through direct free radical scavenging as well as through oxidative hemolysis and lipid peroxidation inhibition in human erythrocytes incubated with an oxidizing agent. In addition, SBSA promoted apoptosis-induced cell death in B16F10Nex-2 by increasing the intracellular reactive oxygen species levels, inducing mitochondrial membrane potential dysfunction and activating caspase-3. Together, these data show the antioxidant and anticancer effects of the aqueous extract from stem bark of Stryphnodendron adstringens, which suggest its potential application for the treatment and/or prevention of diseases related to oxidative stress and tumor cell proliferation.

Evaluation of In Vitro Antioxidant and Anticancer Properties of the Aqueous Extract from the Stem Bark of Stryphnodendron adstringens.

Stryphnodendron adstringens (Mart.) Coville (Fabaceae) is a tree species native to the Brazilian Cerrado commonly known as barbatimão. In traditional medicine, decoctions or infusions of the stem bark of this plant are used in the treatment of several diseases. The objective of this study was to analyze the chemical composition of Stryphnodendron adstringens aqueous extracts (SAAE) prepared from the stem bark to assess their antioxidant activity and anticancer effects as well as characterize cell death mechanisms against murine B16F10Nex-2 melanoma cells. From the SAAE, gallic acid, gallocatechin, epigallocatechin, dimeric and trimeric proanthocyanidins mainly composed of prodelphinidin units and the isomeric chromones C-hexosyl- and O-pentosyl-5,7-dihydroxychromone were identified. The SAAE showed antioxidant activity through direct free-radical scavenging as well as through oxidative hemolysis and lipid peroxidation inhibition in human erythrocytes. Furthermore, SAAE promoted apoptosis-induced cell death in melanoma cells by increasing intracellular reactive oxygen species (ROS) levels, inducing mitochondrial membrane potential dysfunction and activating caspase-3. Together, these data show the antioxidant and anticancer effects of Stryphnodendron adstringens. These results open new perspectives for studies against other tumor cell lines and in vivo models as well as for the identification and isolation of the chemical constituents responsible for these effects.

Abstract 4392: Novel combination of Di-indolylmethane (DIM) and dexamethasone induces apoptosis in myeloma cells by modulating the telomerase activity

Abstract Background and Objectives Multiple myeloma (MM) is a B-cell malignancy involving excessive proliferation and accumulation of atypical plasma cells specifically in the bone marrow (BM) niche. Multiple primary and secondary genetic alterations play crucial role in deregulating the biology of myeloma cells giving them a malignant phenotype, hence generating a need for novel efficacious and safe therapeutic agents. Increased expression of telomerase and higher telomerase activity (RTA) has been found in majority of cancer but displays very low activity in majority of normal somatic cells. Therefore, this maiden attempt unveils the therapeutic potential of Di-indolylmethane alone or in combination with dexamethasone by targeting telomerase activity in myeloma cells. Methodology In this study, two myeloma cell lines (RPMI8226 & U266) were procured from ATCC (Manassas, VA). Di-indolylmethane (DIM) and dexamethasone (Dexa) were purchased from Sigma Aldrich (St. Louis, MO). In in-vitro study, IC50 concentration of DIM and Dexa on both cells was determined using MTT dye. Cell-cycle and apoptosis assay was performed to determine the anti-proliferative and apoptotic effects of DIM and Dexa alone or in combination. Further, caspase-3/7 activity and mitochondrial membrane potential (MMP) was determined to assessed pathway of apoptosis. Effects of single drug or combination on telomerase activity were also assessed in both myeloma cells. Results Results demonstrated that, DIM showed an IC50 conc. of 40μM in both myeloma cells while these cells were resistant to Dexa even at higher dose. In cell cycle assay, significant increase in sub-G0/G1 phase were observed with increasing dose of DIM while combination of DIM and Dexa significantly increased sub-G0/G1 cell populations in comparison to vehicle control. Similarly annexin-V/PI apoptosis assay also demonstrated increased apoptosis of both myeloma cells. Further, DIM alone or in combination with Dexa demonstrated drastic increased caspase-3/7 activity and significant decrease in MMP in both myeloma cells. Additionally, increasing dose of DIM significantly decreased telomerase activity and combination of DIM and Dexa was much more potent in decreasing telomerase activity in comparison to vehicle treatment and DIM alone. Conclusions In nutshell, this maiden attempt provided initial evidences of therapeutic potential of DIM as novel anticancer agent alone or in combination with standard chemotherapeutic agent Dexa. Further, novel combination of DIM and Dexa was much more potential in inducing apoptosis cell death and chemo-sensitizing myeloma cells than single drug treatment regime. Also intrinsic pathway of apoptosis was followed by DIM in inducing apoptosis in myeloma. Furthermore, decreased TA by DIM emphasized its potential as novel molecular therapeutic target which need further validation. Citation Format: Raman Kumar, Nidhi Gupta, . Himani, Alpana Sharma. Novel combination of Di-indolylmethane (DIM) and dexamethasone induces apoptosis in myeloma cells by modulating the telomerase activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4392.

Hydrolysis reaction promotes changes in coordination mode of Ru(II)/acylthiourea organometallic complexes with cytotoxicity against human lung tumor cell lines

In this study, Ru(II)-arene complexes with acylthiourea ligands of the type [Ru(η6‑p‑cymene)(PPh3)(T)Cl]PF6(1–5) and [Ru(η6‑p‑cymene)(PPh3)(T)]PF6(1a, 4a), where PPh3 = triphenylphosphine and T = N‑acyl‑N′(monosubstituted)thiourea, were synthesized and characterized, and their cytotoxic properties were also evaluated. 1a and 4a were obtained from the hydrolysis reaction of 1 and 4. All complexes showed unusual coordination modes for acylthiourea ligands, which are coordinated in a monodentate fashion (S) in 1–5, while they found to be bidentate (S,N), in 1a and 4a. To the best of our knowledge, 1a and 4a are the first crystallographically reported ruthenium compounds with acylthiourea coordinated via S and N(amide) atoms. The cytotoxicity of the compounds was evaluated in human lung cells, A549 and MRC-5. The IC50 values ranging from 0.25 to 0.61 μM after 48 h incubation in lung cancer cells indicate that the compounds showed high cytotoxicity with values significantly lower than the reference drug, cisplatin (11.84 μM). Interaction studies were carried out using human serum albumin (HSA) and DNA. All complexes showed similar cytotoxic activity, however complex 1a, which is the hydrolysis product of 1, presented the highest activity and selectivity among all seven compounds synthesized here. Complexes 1 and 1a inhibited the colony formation decreasing the colony size and inducing morphology changes in A549 cells. These complexes induced apoptosis cell death and promoted cell cycle arrest in the Sub-G1 phase with a decrease in the cell number at the S phase.

Alkaloids from Stephania venosa as Chemo-Sensitizers in SKOV3 Ovarian Cancer Cells via Akt/NF-κB Signaling.

Crebanine (CN), tetrahydropalmatine (THP), O-methylbulbocapnine (OMBC) and N-methyl tetrahydropalmatine (NMTHP) are isoquinoline derived natural alkaloids isolated from tubers of Stephania venosa. We investigated chemo-sensitizing effects of these alkaloids in ovarian cancer cells and evaluated underlying molecular mechanisms involved in chemo-sensitivity. Detection of cell apoptosis was evaluated by using flow cytometry. Cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Chou-Talalay median effect principle was used to evaluate potential drug interactions. Protein analyses were performed on ovarian carcinoma cells using Western blotting upon treatment with anticancer drug and alkaloids. Aporphine alkaloids, such as CN and OMBC, enhanced cisplatin sensitivity in intrinsic cisplatin resistant SKOV3 cells, but not in cisplatin sensitive A2780 cells. Protoberberine alkaloids, such as THP and NMTHP, had no synergistic effect on cisplatin sensitivity in either cell line. Chemo-sensitizing effects of CN and OMBC in SKOV3 cells were mediated via activating apoptosis-induced cell death through caspase-3, -8 and cleaved poly ADP-ribose polymerase (PARP) and via inhibiting anti-apopotic and survival protein expression, such as Bcl-xL, Baculoviral IAP repeat-containing protein 3 (cIAP-2), survivin and interleukin (IL) -6. Cisplatin stimulated protein kinase B (Akt) and nuclear factor-kappaB (NF-κB) signaling pathways, but not mitogen-activated protein kinase (MAPK), activator protein 1 (AP-1) and signal transducer and activator of transcription 3 (STAT3) in SKOV3 cells. Akt/NF-κB signaling was blocked by CN and OMBC leading to increased sensitization to cisplatin. These findings demonstrate that CN and OMBC sensitizes SKOV3 cells to cisplatin via inhibition of Akt/NF-κB signaling and the down regulation of NF-κB mediated gene products. Our results suggest that alkaloids obtained from S. venosa could be used as chemo-sensitizers in ovarian cancer to sensitize and minimize the dose related toxicity of platinum-based chemotherapeutic drugs.

Ficus virens proanthocyanidins induced apoptosis in breast cancer cells concomitantly ameliorated 5-fluorouracil induced intestinal mucositis in rats

5-Fluorouracil (5-FU) is a commonly used chemotherapeutic agent for breast cancer. However, its use often leads to drug resistance and mucositis. This study aimed to investigate whether proanthocyanidins from Ficus virens possessed anti-breast cancer and anti-mucositis activities. The results showed that the cytotoxic effects of the proanthocyanidins against MDA-MB-231 and MCF-7 breast cancer cells were in the order of stem barks proanthocyanidins (SPAs) > leaves proanthocyanidins > fruits proanthocyanidins. Moreover, SPAs induced apoptosis in both cell lines which were accompanied with an increase in loss of mitochondrial membrane potential, production of reactive oxygen species, Bax to Bcl-2 protein expression ratio, and activated caspase 3. Furthermore, intraperitoneal injection of 5-FU (150 mg/kg body weight) resulted in body weight loss and jejunal injury in the rats while administration of SPAs (100 mg/kg body weight) counteracted these changes. Collectively, our study demonstrated that SPAs induced apoptosis cell death in breast cancer cells while ameliorating the symptoms of intestinal mucositis in rats.Therefore, SPAs merits further exploration as a potential therapeutic agent for breast cancer and chemotherapy-induced mucositis.

Antiproliferative effect of apocynin in cervical epithelial cells infected by HPV 16 involves change of ROS production and cell cycle

The cervical cancer is characterized as a public health problem. The human papillomavirus, especially the high-risk types 16 and 18, is present in 99.7% of cases of invasive cervical cancers. Due to high prevalence, the search for new drugs for the treatment is continuous. The apocynin, compound belonging to the class of catechol-methoxy acetophenone, has been used as an effective and non-toxic inhibitor of the NADPH oxidase complex, in many experimental models. Some early studies have indicated that apocynin also exerts antiproliferative activity inducing cell death by apoptosis. Thus, we evaluated the effect of apocynin in cervical epithelial cells infected by human papillomavirus 16 (SiHa cells) by analyzing oxygen species production, cell cycle, cell morphology, cell volume, membrane integrity and mitochondria membrane potential. Our data showed that apocynin induces antiproliferative effects by decreasing reactive oxygen species production and inducing cell cycle arrest, activating pathways that induce apoptosis cell death.

Effects of low dose of bisphenol A on the proliferation and mechanism of primary cultured prostate epithelial cells in rodents.

Bisphenol A (BPA) is a well-known endocrine disruptor compound (EDC) that aggravates testosterone-induced benign prostate hyperplasia by increasing the relative weight of the ventral and dorsolateral prostate in rats. This phenomenon is primarily attributed to the exogenous estrogen effect of BPA. However, the direct effect of BPA on prostate cells has not been characterized. The present study investigated the proliferative effect and possible mechanisms of action of BPA on the prostatic epithelium of rats. The ventral prostate epithelial cells were cultured in vitro and the proliferation effects of BPA on cells were studied. The cells were identified as prostatic epithelial cells, and cell viability, cell apoptosis and the expressions of androgen receptors (AR) and estrogen receptors (ER), were detected. It was observed that 0.01–1 nM BPA promoted cell growth, with 1 nM BPA inducing the greatest increase in the rate of cell growth. However, BPA-treated cells exhibited no marked morphological changes compared with the control group. The cell apoptosis rate in each BPA-treated group was lower compared with the control group. The expression levels of ERα and ERβ increased, but the expression of AR decreased. The present study demonstrated that environmental exposure to BPA directly promoted the proliferation of prostate cells in rats through increasing the expression of estrogen receptors, reducing the expression of androgen receptors of the cells and decreasing apoptosis-induced cell death.

Stereochemical preference toward oncotarget: Design, synthesis and in vitro anticancer evaluation of diastereomeric β-lactams.

PurposeIn the battle against cancer discovery of new and novel chemotherapeutic agent demands extreme obligation. Development of anticancer compounds with higher potency and reduced side-effects is timely and challenging.Experimental DesignA small series of fourteen diastereomeric β-lactams (seven pairs) were synthesized through multi-step process exploring [2+2] ketene-imine cycloaddition as the key step. Comparative stereochemical preferences were studied through computational docking and validated by in vitro evaluation. β-tubulin was considered as possible molecular target and in vitro anticancer evaluation was conducted against SiHa, B16F10, K562 and Chang cell lines. Caspase-3 activation assay and hematoxylin/eosin staining of the cells were also accomplished.ResultsBetter docking scores of the cis- over the trans-β-lactams indicated favorable β-lactam—β-tubulin interactions in cis-geometry. In vitro (IC50) evaluation confirmed better anticancer activity of the cis-diastereoisomers. Apoptosis-induced cell death was supported by caspase-3 activation study. A cis-β-lactam [(±)-Cis-3-amino-1-phenyl-4-(p-tolyl) azetidin-2-one, 6C] was found to be more active (in vitro) than the marketed natural drug colchicine against SiHa and B16F10 (six times higher potency) cell lines. Reduced toxicity (compared to colchicine) in Chang cells confirmed better site-selectivity (accordingly less side-effects) of 6C than colchicine. Aside from 6C, most of the reported molecules demonstrated good to strong in vitro anticancer activity against SiHa and B16F10 cancer cell lines.ConclusionsStereochemical preferences of the cis-β-lactams over their trans-counterparts, toward the molecular target β-tubulin, was confirmed by docking studies and in vitro anticancer evaluation. Apoptosis was identified as the cause of cell death. The lead 6C exhibited higher potency and selectivity than the marketed drug colchicine both in silico as well as in vitro.

Antiproliferative and Apoptotic Effects of Novel Anti-ROR1 Single-Chain Antibodies in Hematological Malignancies

Receptor tyrosine kinase–like orphan receptor (ROR) proteins are a conserved family of tyrosine kinase receptors that function in developmental processes including cell survival, differentiation, cell migration, cell communication, cell polarity, proliferation, metabolism, and angiogenesis. ROR1 has recently been shown to be expressed in various types of cancer cells but not normal cells. Pharmacokinetics and pharmacodynamics of single-chain Fragment variable (scFv) antibodies provide potential therapeutic advantages over whole antibody molecules. In the present study, scFvs against a specific peptide from the extracellular domain of ROR1 were selected using phage display technology. The selected scFvs were further characterized using polyclonal and monoclonal phage enzyme-linked immunosorbent assay (ELISA), soluble monoclonal ELISA, colony PCR, and sequencing. Antiproliferative and apoptotic effects of selected scFv antibodies were also evaluated in lymphoma and myeloma cancer cell lines using MTT and annexin V/PI assays. The results of ELISA indicated specific reactions of the isolated scFvs against the ROR1 peptide. Colony PCR confirmed the presence of full-length VH and Vκ inserts. The percentages of cell growth after 24 h of treatment of cells with individual scFv revealed that the scFv significantly inhibited the growth of the RPMI8226 and chronic lymphocytic leukemia (CLL) cells in comparison with the untreated cells (p < 0.05). Interestingly, 24-h treatment with specific scFv induced apoptosis cell death in the RPMI8226 and CLL cells. Taken together, our results demonstrate that targeting of ROR1 using peptide-specific scFv can be an effective immunotherapy strategy in hematological malignancies.

Phaleria macrocarpa (Boerl.) fruit induce G0/G1 and G2/M cell cycle arrest and apoptosis through mitochondria-mediated pathway in MDA-MB-231 human breast cancer cell

Ethnopharmacological relevancePhaleria macrocarpa (Scheff) Boerl, is a well-known folk medicinal plant in Indonesia. Traditionally, P. macrocarpa has been used to control cancer, impotency, hemorrhoids, diabetes mellitus, allergies, liver and hearth disease, kidney disorders, blood diseases, acne, stroke, migraine, and various skin diseases. Aim of the studyThe purpose of this study was to determine the in situ cytotoxicity effect P. macrocarpa fruit ethyl acetate fraction (PMEAF) and the underlying molecular mechanism of cell death. Materials and methodsMDA-MB-231 cells were incubated with PMEAF for 24h. Cell cycle and viability were examined using flow cytometry analysis. Apoptosis was determined using the Annexin V assay and also by fluorescence microscopy. Apoptosis protein profiling was detected by RayBio® Human Apoptosis Array. ResultsThe AO/PI staining and flow cytometric analysis of MDA-MB-231 cells treated with PMEAF were showed apoptotic cell death. The cell cycle analysis by flow cytometry analysis revealed that the accumulation of PMEAF treated MDA-MB-231 cells in G0/G1 and G2/M-phase of the cell cycle. Moreover, the PMEAF exert cytotoxicity by increased the ROS production in MDA-MB-231 cells consistently stimulated the loss of mitochondrial membrane potential (∆Ψm) and induced apoptosis cell death by activation of numerous signalling proteins. The results from apoptosis protein profiling array evidenced that PMEAF stimulated the expression of 9 pro-apoptotic proteins (Bax, Bid, caspase 3, caspase 8, cytochrome c, p21, p27, p53 and SMAC) and suppressed the 4 anti-apoptotic proteins (Bcl-2, Bcl-w, XIAP and survivin) in MDA-MB-231 cells. ConclusionThe results indicated that PMEAF treatment induced apoptosis in MDA-MB-231 cells through intrinsic mitochondrial related pathway with the participation of pro and anti-apoptotic proteins, caspases, G0/G1 and G2/M-phases cell cycle arrest by p53-mediated mechanism.

Augmentation of poly(ADP-ribose) polymerase-dependent neuronal cell death by acidosis

Tissue acidosis is a key component of cerebral ischemic injury, but its influence on cell death signaling pathways is not well defined. One such pathway is parthanatos, in which oxidative damage to DNA results in activation of poly(ADP-ribose) polymerase and generation of poly(ADP-ribose) polymers that trigger release of mitochondrial apoptosis-inducing factor. In primary neuronal cultures, we first investigated whether acidosis per sé is capable of augmenting parthanatos signaling initiated pharmacologically with the DNA alkylating agent, N-methyl-N′-nitro-N-nitrosoguanidine. Exposure of neurons to medium at pH 6.2 for 4 h after N-methyl-N′-nitro-N-nitrosoguanidine washout increased intracellular calcium and augmented the N-methyl-N′-nitro-N-nitrosoguanidine-evoked increase in poly(ADP-ribose) polymers, nuclear apoptosis-inducing factor , and cell death. The augmented nuclear apoptosis-inducing factor and cell death were blocked by the acid-sensitive ion channel-1a inhibitor, psalmotoxin. In vivo, acute hyperglycemia during transient focal cerebral ischemia augmented tissue acidosis, poly(ADP-ribose) polymers formation, and nuclear apoptosis-inducing factor , which was attenuated by a poly(ADP-ribose) polymerase inhibitor. Infarct volume from hyperglycemic ischemia was decreased in poly(ADP-ribose) polymerase 1-null mice. Collectively, these results demonstrate that acidosis can directly amplify neuronal parthanatos in the absence of ischemia through acid-sensitive ion channel-1a . The results further support parthanatos as one of the mechanisms by which ischemia-associated tissue acidosis augments cell death.

Chitosan-based nano-in-microparticle carriers for enhanced oral delivery and anticancer activity of propolis

This study reports a promising approach to enhance the oral delivery of propolis, improve its aqueous solubility and bioavailability, and allow its controlled release as well as enhancing its anticancer activity. Propolis was standardized then its solubility was improved via formulation into optimized solid dispersion (SD) matrices, and its release was controlled through incorporation into nanoparticles (NPs) of optimized composition followed by further inclusion into chitosan (Cs) microparticles. The anticancer activity of the newly developed propolis-loaded nano-in-microparticles (NIMs) was evaluated against human liver cancer (HepG2) and human colorectal cancer (HCT 116) cells. The prepared SDs, NPs and NIMs were characterized using SEM, TEM, DLS, FTIR, DSC and UV⿿vis spectrophotometry. The therapeutic efficiency of formulated propolis was bio-assessed via cytotoxicity measurements, mitochondrial dysfunction, apoptosis-induced cell death and cell cycle arrest. The results demonstrated a considerable enhancement in propolis solubility with a controlled release profile in different GIT environments. In-vitro cytotoxicity studies showed that the propolis-loaded NIMs induce more cytotoxic effect on HepG2 cells than HCT-116 cells and mediated three-fold higher therapeutic efficiency than free propolis. The apoptosis assay indicated that the propolis-loaded NIMs induce apoptosis of HepG2 cells and significantly decrease their number in the proliferative G0/G1, S and G2/M phases.

Aristolochic Acid I Causes Testis Toxicity by Inhibiting Akt and ERK1/2 Phosphorylation

Aristolochic acid (AA) is a natural bioactive substance found in Chinese herbs that induce toxicity during ovarian maturation of animals and humans. Apoptosis is induced by various types of damage and governs the progression of biological cell removal that controls the equilibrium between cell growth and death. However, the AA toxicity mechanism during testis maturation in mouse has not been elucidated and was thus the focus of the present study. This study used TM4 Sertoli cells and an ICR mouse model, both of which were injected with aristolochic acid I (AAI) for 4 weeks. Testis dimensions and weight were surveyed to define AAI cytotoxicity in the mice testis. The MTT assay was used to analyze the cytotoxicity of AAI in TM4 Sertoli cells. An apoptosis expression mediator was analyzed through Western blotting, while the measure of apoptosis-induced cell death of TM4 Sertoli cells and testis tissues was analyzed by the TUNEL assay. We found that AAI strongly inhibits survival in TM4 cells and that AAI significantly activated apoptosis-induced cell death in TM4 Sertoli cells and mice testis tissue. In addition, AAI suppressed the expression of B-cell lymphoma 2 (Bcl-2), a factor related to anti-apoptosis. It markedly improved pro-apoptotic protein expression, including Bcl-2-associated X protein, poly(ADP-ribose) polymerase, and caspase-3 and -9. Furthermore, we observed that AAI significantly reduced the size and weight of mouse testis. Moreover, germ cells and somatic cells in testis were markedly damaged by AAI. In addition, we found that AAI significantly inhibits ERK1/2 and Akt activation in TM4 Sertoli cells and testis tissue. The data obtained in this study indicate that AAI causes severe injury for the period of testis development by impeding apoptosis related to the Akt and ERK1/2 pathway.

백굴채 추출물이 MDA-MB-231 유방암 세포주에서 세포사멸에 미치는 효과

ABSTRACT Effects of Chelidonium Majus Extract on Apoptosis Induction of MDA-MB-231 Human Breast Cancer CellsSae-Byul Jang, Dong-Youl YooDept. of Oriental Gynecology, College of Oriental Medicine,Dae-Jeon UniversityObjectives: In this study, we investigated the anti-proliferative and apoptosis inducing effect of water extract of Chelidonium majus (CM) on human breast cancer cell MDA-MB-231.Methods: The MTT assay was used to assess cell proliferation. The expre ssion of apoptosis related gene was assessed by quantitative Real-tim e PCR. Cell apoptosis detected by flow cytometry using Annexin-V/PI staining.Results: Our data revealed that CM inhibited the cell growth in a dose dependent manner (0, 62.5, 125, 250, 500 μg/ml). CM increased mRNA expression of pro-apoptotic genes Bax, caspase-3, and caspase-9. Annexin-V/PI staining assa ys revealed that apoptosis-induced cell death increased in a dose-dependent mann er in cells.Conclusions: CM induces cell death in MDA-MB-231 human breast cancer cell and shows potentials for use in cancer therapy as apoptosi s-inducing agent.Key Words: Chelidonium majus , breast cancer, MDA-MB-231, apoptosis, anti-proliferative

Rhodium Citrate Associated with Maghemite Nanoparticles Causes DNA Fragmentation Independently of Caspases 3 and Mediated by Reactive Oxygen Species

Breast cancer is the most common cancer among women, excluding non-melanoma skin cancer. Research efforts have been directed towards the development of more efficient drugs against this disease, such as metal complexes, which have been widely studied. These compounds can intercalate in DNA bases and impair DNA transcription and replication, leading to cell death. Cell death can also be associated with early induction of reactive oxygen species (ROS) production by cells treated with this kind of metal complex. Nevertheless, the use of these compounds is limited because of their systemic toxicity. In this regard, the use was proposed of dirhodium citrate [Rh2(H2cit)4] associated with magnetic nanoparticles (NPs), which are carriers that may work in decreasing systemic toxicity. We compared cell viability effects of free Rh2(H2cit)4, Rh2(H2cit)4-loaded maghemite NPs [Magh-Rh2(H2cit)4] and maghemite nanoparticles loaded with citrate (Magh-cit), on MCF-7 breast cancer cells and MCF-10A non-tumor and non-tumorigenic epithelial cells by MTT assay. Furthermore, we examined whether the NPs induce cell death by apoptosis in a cell line without caspase 3 expression (MCF-7). This feature was demonstrated by quantification of ROS through labeling cells with DCFDA, DNA fragmentation studies analyzed with a flow cytometer, release of cytochrome C from mitochondria assays, and effector caspases activation analysis (revealed by FLICA) as visualized by confocal microscopy. Our results confirmed that rhodium citrate was less cytotoxic in its free form than when associated with the tested drug delivery system. Moreover, Magh-Rh2(H2cit)4 NPs and Magh-cit NPs induced apoptosis cell death mediated by ROS and independently of caspase 3 expression.

Oligonol, a low-molecular-weight polyphenol derived from lychee fruit, attenuates gluco-lipotoxicity-mediated renal disorder in type 2 diabetic db/db mice.

Oligonol is a phenolic product derived from lychee fruit extract containing catechin-type monomers and oligomers of proanthocyanidins, produced by a manufacturing process which converts polyphenol polymers into oligomers. These proanthocyanidins have been reported to exhibit beneficial bioactivities in many studies, and so oligonol, a rich source of polyphenol, is expected to show favorable effects on various chronic diseases. This article summarizes recent work whether oligonol has an ameliorative effect on diabetic indices and renal disorders associated with gluco-lipotoxicity-mediated oxidative stress, inflammation, and apoptosis in db/db mice with diabetes. Oligonol was able to improve diabetic indices, prevent the development of diabetic renal disease, and preserve renal cells and the renal morphological structure via the attenuation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-induced oxidative stress, inhibition of advanced glycation endproduct (AGE) generation, and prevention of apoptosis-induced cell death in db/db mice, being independent of changes in the body weight or serum glucose levels. The present study provides important evidence that oligonol exhibits a pleiotropic effect, representing renoprotective effects against the development of diabetic complications in type 2 diabetic db/db mice.

Abstract 5118: Decursin synergistically enhances TRAIL-induced apoptosis through CHOP dependent DR5 signaling in TRAIL resistant lung cancer cells

Abstract Targeting the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), which has been clinically established for anticancer therapy, is the attractive strategy to specifically kill cancer cells through the death receptors. Nonetheless, its therapeutic potential is limited due to the resistance in cancer cells. Thus, in the present study, we evaluated the therapeutic potential of Decursin, a pyranocoumarin isolated from the Korean Angelica gigas root as a TRAIL sensitizer in a variety of resistant lung cancer cells. Co-treatment of Decursin with TRAIL synergistically induced apoptosis in TRAIL resistant lung cancer cells, A545, H1299, H596 and Calu-1. This synergistic effect through the induction of DR5 in Decursin treated cells was also confirmed by immunoblotting and immunoanalysis of endogeneous and surface expression of DR5, respectively. In contrast, in DR knockdown cells, combination treatment Decursin and TRAIL did not induce apoptosis at all. Furthermore, we found that DR5 induction by Decursin was mediated by ER-stress and its byproduct CHOP. Depletion of CHOP via siRNA transfection completely blocked Decusin induced apoptosis cell death in TRAIL resistant lung cancer cells. Taken together, induction of the DR5 via upregulation of CHOP and its inducer, ER stress, by Decursin leads to synergistically increase the TRAIL mediated cell death in TRAIL resistant lung cancer cells. Our results indicate that Decursin is a useful therapeutic strategy for overcoming TRAIL resistance in lung tumor treatment. Key words: Decursin, DR-5, ER stress, CHOP, Trail resistant, Lung cancer Conflict of Interests: The authors disclose no potential conflicts of interest. Acknowledgment: This work was supported by the National Research Foundation of Korea (NRF) Grant funded by the Korea Government (MEST) (no. 2012-0005755). Citation Format: Kim Jae Kwang. Decursin synergistically enhances TRAIL-induced apoptosis through CHOP dependent DR5 signaling in TRAIL resistant lung cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5118. doi:10.1158/1538-7445.AM2014-5118

Gold(I) complexes with alkylated PTA (1,3,5-triaza-7-phosphaadamantane) phosphanes as anticancer metallodrugs

New stable thiolate gold(I) derivatives containing the alkylated phosphanes [PTA–CH2Ph]Br and [PTA–CH2COOMe]Br derived from 1,3,5-triaza-7-phosphaadamantane (PTA) have been prepared by different routes of synthesis. By the use of basic media to deprotonate the corresponding thiol in the former and by transmetallation reactions from tin (IV) complexes, in the later, thus avoiding side reactions on the phosphane. Strong antiproliferative effects are observed for most of the compounds, including the chloro- and bromo precursors with the series of phosphanes derived from PTA, in human colon cancer cell lines (Caco-2, PD7 and TC7 clones). Apoptosis-induced cell death is found for all compounds, being the thiolate derivatives with [PTA–CH2Ph]Br the most effective, as shown by an annexin-V/propidium iodide double-staining assay.