Inductor Of Apoptosis Research Articles

Evaluating genotoxic risks in Brazilian public health agents occupationally exposed to pesticides: a multi-biomarker approach.

This is the first study demonstrating genotoxic effects and whole transcriptome analysis on community health agents (CHAs) occupationally exposed to pesticides in Central Brazil. For the transcriptome analysis, we found some genes related to Alzheimer's disease (LRP1), an insulin-like growth factor receptor (IGF2R), immunity genes (IGL family and IGJ), two genes related to inflammatory reaction (CXCL5 and CCL3), one gene related to maintenance of cellular morphology (NHS), one gene considered to be a strong apoptosis inductor (LGALS14), and several transcripts of the neuroblastoma breakpoint family (NBPF). Related to comet assay, we demonstrated a significant increase in DNA damage, measured by the olive tail moment (OTM), in the exposed group compared to the control group. Moreover, we also observed a statistically significant difference in OTM values depending on GSTM1 genotypes. Therefore, Brazilian epidemiological surveillance, an organization responsible for the assessment and management of health risks associated to pesticide exposure to CHA, needs to be more proactive and considers the implications of pesticide exposure for CHA procedures and processes.

Evaluation of Polybrominated Diphenyl Ether Toxicity on HepG2 Cells – Hexabrominated Congener (BDE‐154) Is Less Toxic than Tetrabrominated Congener (BDE‐47)

Apoptotic cell death is one of the main consequences of exposure to brominated flame retardants, including polybrominated diphenyl ethers. However, few of these compounds have had their potential toxicity investigated. BDE-154 is one of the most poorly studied polybrominated diphenyl ether (PBDE) congeners, but its level in the environment and in biological fluids is rising. In addition, its chemical structure differs from the other congeners with well-documented toxicity, so BDE-154 may display a distinct toxicity pattern. This study has evaluated how BDE-154 affects the human hepatoblastoma cell line (HepG2) and has looked into the impact of this congener on human health. In addition, this study has related the effects of BDE-154 with the effects of BDE-47 to clarify the mechanism of PBDE toxicity. The HepG2 cell line was exposed to BDEs for 24 and 48 hr and submitted to assays to examine proliferation, viability, mitochondrial membrane potential, reactive oxygen species accumulation, phosphatidylserine exposure, nuclear fragmentation and evaluation of pro-caspase 3, pro-caspase 9, cytochrome c release, and apoptosis inductor factor release by Western blot analysis. BDE-154 induced mitochondrial damage and led to apoptotic death of HepG2 cells, but these effects were less intense than the effects promoted by BDE-47. Unlike other extensively reported congeners, BDE-154 was only toxic at the higher tested concentrations, whereas BDE-47 cytotoxicity was evident even at lower concentrations. Hence, like the toxicity pattern of other classes of substances such as polychlorinated biphenyls, the toxicity pattern of BDEs also depends on their chemical structure and aromatic substituent.

Study of Association between Pre-Senile Cataracts and the Polymorphisms rs2228000 in XPC and rs1042522 in p53 in Spanish Population.

PurposeTo determine if the presence of certain polymorphisms in the DNA repair gene XPC and the apoptosis inductor gene p53 is associated with pre-senile cataract development.MethodsWe have performed a retrospective study over three groups of patients. The group with pre-senile cataract formed by 72 patients younger than 55 with cataract surgery. The group with senile cataract formed by 101 patients older than 55 with cataract surgery. The group without cataract was formed by 42 subjects older than 55 without lens opacities. We analyzed the presence of SNP rs2228000 from XPC and rs1042522 from p53; and the relationship between risk factors such as smoking, alcohol intake, hypertension or diabetes.ResultsThe comparison of the genotype distribution in XPC, within the different groups, did not show any statistically significant association in any of our analysis (p>0,05). The comparison of the genotype distribution in p53 within the different groups did not show any statistically significant association (p>0,05); except for the comparison between the pre-senile cataract group and the group with senile cataract where the genotype Pro/Pro (C/C) in the recessive inheritance model showed a higher risk for developing pre-senile cataract (p = 0,031; OR = 1.04–15.97). This association decreased when we performed the analysis adjusting by the studied risk factors (p = 0.056).ConclusionsAllelic variants in the gene XPC are not associated with an increased risk for developing pre-senile cataract. The presence of the genotype Pro/Pro in p53 might be associated with a major risk for developing pre-senile cataract.

In Vitro Study of Cytophysiological Characteristics of Multinuclear Macrophages from Intact and BCG-Infected Mice.

Peritoneal macrophages were isolated from intact and BCG-infected BALB/c mice and explanted in vitro. Multinuclear macrophages formed in these cultures differed by the number of nuclei, expression of apoptosis inductors and regulators (TNF-α, p53 protein, caspase 3, and Bcl-2 protein), and cytophysiological characteristics (phagocytic activity, ROS generation, and antimycobacterial properties). Our results indicate that the formation of multinuclear macrophages is accompanied by induction of apoptosis (p53 signaling pathway) and appearance of multinuclear macrophage-derived cells characterized by high phagocytic and antimycobacterial activity.

Fast Centrifugal Partition Chromatography Fractionation of Concentrated Agave (Agave salmiana) Sap to Obtain Saponins with Apoptotic Effect on Colon Cancer Cells.

Separation of potentially bioactive components from foods and plant extracts is one of the main challenges for their study. Centrifugal partition chromatography has been a successful technique for the screening and identification of molecules with bioactive potential, such as steroidal saponins. Agave is a source of steroidal saponins with anticancer potential, though the activity of these compounds in concentrated agave sap has not been yet explored. In this study, fast centrifugal partition chromatography (FCPC) was used coupled with in vitro tests on HT-29 cells as a screening procedure to identify apoptotic saponins from an acetonic extract of concentrated agave sap. The three most bioactive fractions obtained by FCPC at partition coefficients between 0.23 and 0.4 contained steroidal saponins, predominantly magueyoside b. Flow cytometry analysis determined that the fraction rich in kammogenin and manogenin glycosides induced apoptosis, but when gentrogenin and hecogenin glycosides were also found in the fraction, a necrotic effect was observed. In conclusion, this study provides the evidence that steroidal saponins in concentrated agave sap were potential inductors of apoptosis and that it was possible to separate them using fast centrifugal partition chromatography.

Analysis of Tumor Necrosis Factor Function Using the Resonant Recognition Model.

The tumor necrosis factor (TNF) is a complex protein that plays a very important role in a number of biological functions including apoptotic cell death, tumor regression, cachexia, inflammation inhibition of tumorigenesis and viral replication. Its most interesting function is that it is an inhibitor of tumorigenesis and inductor of apoptosis. Thus, the TNF could be a good candidate for cancer therapy. However, the TNF has also inflammatory and toxic effects. Therefore, it would be very important to understand complex functions of the TNF and consequently be able to predict mutations or even design the new TNF-related proteins that will have only a tumor inhibition function, but not other side effects. This can be achieved by applying the resonant recognition model (RRM), a unique computational model of analysing macromolecular sequences of proteins, DNA and RNA. The RRM is based on finding that certain periodicities in distribution of free electron energies along protein, DNA and RNA are strongly correlated to the biological function of these macromolecules. Thus, based on these findings, the RRM has capabilities of protein function identification, prediction of bioactive amino acids and protein design with desired biological function. Using the RRM, we separate different functions of TNF as different periodicities (frequencies) within the distribution of free energy electrons along TNF protein. Interestingly, these characteristic TNF frequencies are related to previously identified characteristics of proto-oncogene and oncogene proteins describing TNF involvement in oncogenesis. Consequently, we identify the key amino acids related to the crucial TNF function, i.e. receptor recognition. We have also designed the peptide which will have the ability to recognise the receptor without side effects.

Abstract 2914: Gold magneto nanoparticles induced apoptosis in liver cancer cells through changes of Ca channel pump detected by LIBS Technique and RT-PCR array

Abstract Laser-induced breakdown spectroscopy technique (LIBS) was used in thesis study to address the effect of gold magneto nanoparticles on the liver cancer cells (HepG2) by examining the variation in genotoxicity and cytotoxicity. HepG2 cell line was cultured at 105 cells/ml and expanded, after which, they were incubated with gold magneto nanoparticles at concentrations 10, 25, 50 μg/ml respectively for 12 hours. The effect of these nanoparticles in cell apoptosis as well as in autophagy was evaluated by acridine orange staining and Annexin V/propidium iodide double staining as well as apoptosis specific PCR array assay for evaluation of apoptosis in the treated cells. At a concentration of nanocomposite 10, 25, 50 μg/ml, cells showed increase in the apoptosis figure from 11.25% ± 3.1% to 28% ± 2.04%, 39% ± 3.6%, 60% ± 4.3% respectively. Apoptosis specific PCR array also, showed very high expression of TNF, BIRC3 and NFKB1 with increasing in the fold regulation ratio of 155.03,158.03 and 149.2- fold respectively. On the other hand, extra investigation of these treated cells by LIBS shows an increasing in the iron oxide nanoparticles concentration as well as calcium concentration. This increase in the calcium content in the treated cells may play an important role in the program cell apoptosis which was prove in our study by both Flow cytometry as well as by profile apoptosis PCR array. Finally, our data showed the possibility of the using LIBS as inductor of cells apoptosis by measurement Ca concentration changes in liver cancer cells (HepG2 cells). Keywords: Apaptosis, LIBS; HepG2 cell; Genotoxicity Citation Format: Ola Ahmed, Hisham Imam, Abdel-Rahman N. Zekri. Gold magneto nanoparticles induced apoptosis in liver cancer cells through changes of Ca channel pump detected by LIBS Technique and RT-PCR array. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2914. doi:10.1158/1538-7445.AM2015-2914

Influence of the Paracoccidioides brasiliensis 14-3-3 and gp43 proteins on the induction of apoptosis in A549 epithelial cells.

The fungal strain Paracoccidioides brasiliensis remains viable inside of epithelial cells and can induce apoptosis in this population. However, until now, the molecules that participate in this process remained unknown. Thus, this study evaluated the contribution of two P. brasiliensis molecules, the 14-3-3 and glycoprotein of 43 kDa proteins, which had been previously described as extracellular matrix adhesins and apoptosis inductors in human pneumocytes. Accordingly, epithelial cells were treated with these molecules for different periods of time and the expression of the apoptosis regulating-proteins Bak, Bax, Bcl-2, p53 and caspases were evaluated by terminal deoxynucleotidyl transferase dUTP nick end labelling, flow cytometry and real-time polymerase chain reaction analysis. Our results demonstrated that treatment with these molecules induces apoptosis signalling in pulmonary epithelial cells, showing the same pattern of programmed cell-death as that observed during infection with P. brasiliensis. Thus, we could conclude that P. brasiliensis uses these molecules as virulence factors that participate not only in the fungal adhesion process to host cells, but also in other important cellular mechanisms such as apoptosis.

THE CHARACTERIZATION OF PROAPOPTOTIC ACTIVITY OF LIPOPOLYSACCHARIDE IN ISOLATED SUBPOPULATIONS OF HUMAN T-LYMPHOCYTES

Lipopolysaccharide (LPS) is used as activator and inductor of immune cells apoptosis in case of infectious processes modeling. It is not clear if LPS is direct inductor of apoptosis in human T-cells. The subpopulations of effector and naive CD4+ T-helper and CD8+ cytotoxic T lymphocytes of human peripheric blood were isolated and cultivated in presence of LPS. It has been shown that LPS was inductor of apoptosis in studying T-cells populations.

Abstract 3791: The inhibition of protein kinase CK2 activity by the CIGB-300 synthetic peptide impairs the three-dimensional cell growth, Wnt and nuclear factor қB signaling pathways in lung cancer cells

Abstract CK2 is a serine/threonine kinase involved in cell growth, survival and apoptosis. The CIGB-300 is a synthetic peptide capable of binding to CK2 substrates thus preventing the enzyme activity. Previously we have determined that CIGB-300 presented an inhibitory concentration 50 dose (IC50) of 119±2.4 µM in NCI-H125 human lung cancer cells. Throughout an Annexin V-FITC assay we could determine that CIGB-300 is a potent apoptosis inductor since the treatment with this drug for 60 minutes induced apoptosis at a level comparable with that observed with etoposide (10 µM). Concomitantly with cell death induction, we could observe caspase-3 activation and the decreased expression of c-myc and cyclin D1 and D2. In this work we analyzed whether CIGB-300 is able to alter the ability of cells to grow in 3D and to modulate signaling pathways involved in tumor progression. First, we developed stable spheroids of NCI-H125 cells, which were able to grow in culture for at least 14 days. After 5 days of treatment, CIGB-300 induced a significant growth inhibition (median volume: 4.07x107±0.70x107 µm3 in control spheroids vs 2.13x107±0.35x107 µm3 in spheroids treated with the IC50 dose p<0.05). From the sixth day of treatment the size of the spheroids was dramatically reduced, returning to the initial volume on day eight. To analyze the effect of CIGB-300 on canonical Wnt signaling, this pathway was initially activated by incubating NCI-H125 cells with a conditioned media containing Wnt3a factor. This incubation led to a significant increase in the levels of cytoplasmic β-catenin. By Western blot we could observe that treatment with CIGB-300 blocked this increase, suggesting its role in the modulation of the Wnt pathway. In order to analyze the nuclear factor κB (NF-κB) dependent pathway, this transcription factor was activated by a phorbol ester treatment (PMA, 15 nM). Short treatment (15 min) with CIGB-300 induced an important reduction in NF-κB nuclear levels. However, this inhibition was reverted 24 h later as determined by Western blot and reporter gene expression assays. Our results indicate that the treatment with CIGB-300 induces a significant anti-proliferative response in a three-dimensional model, which resembles in vivo tumor growth conditions, also affecting key signaling pathways involved in tumor progression. Altogether, our data suggest that this peptide may become a new strategy for the treatment of lung cancer malignancies. Citation Format: Stéfano M. Cirigliano, María Inés Díaz Bessone, Carolina Flumian, Damián Emilio Berardi, Silvio E. Perea, Elisa Bal De Kier Joffé, Hernán Farina, Laura Todaro, Alejandro Urtreger. The inhibition of protein kinase CK2 activity by the CIGB-300 synthetic peptide impairs the three-dimensional cell growth, Wnt and nuclear factor қB signaling pathways in 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 3791. doi:10.1158/1538-7445.AM2014-3791

Modeling pulmonary fibrosis by abnormal expression of telomerase/apoptosis/collagen V in experimental usual interstitial pneumonia.

Limitations on tissue proliferation capacity determined by telomerase/apoptosis balance have been implicated in pathogenesis of idiopathic pulmonary fibrosis. In addition, collagen V shows promise as an inductor of apoptosis. We evaluated the quantitative relationship between the telomerase/apoptosis index, collagen V synthesis, and epithelial/fibroblast replication in mice exposed to butylated hydroxytoluene (BHT) at high oxygen concentration. Two groups of mice were analyzed: 20 mice received BHT, and 10 control mice received corn oil. Telomerase expression, apoptosis, collagen I, III, and V fibers, and hydroxyproline were evaluated by immunohistochemistry, in situ detection of apoptosis, electron microscopy, immunofluorescence, and histomorphometry. Electron microscopy confirmed the presence of increased alveolar epithelial cells type 1 (AEC1) in apoptosis. Immunostaining showed increased nuclear expression of telomerase in AEC type 2 (AEC2) between normal and chronic scarring areas of usual interstitial pneumonia (UIP). Control lungs and normal areas from UIP lungs showed weak green birefringence of type I and III collagens in the alveolar wall and type V collagen in the basement membrane of alveolar capillaries. The increase in collagen V was greater than collagens I and III in scarring areas of UIP. A significant direct association was found between collagen V and AEC2 apoptosis. We concluded that telomerase, collagen V fiber density, and apoptosis evaluation in experimental UIP offers the potential to control reepithelization of alveolar septa and fibroblast proliferation. Strategies aimed at preventing high rates of collagen V synthesis, or local responses to high rates of cell apoptosis, may have a significant impact in pulmonary fibrosis.

Activity of 4-arylchromene derivatives as apoptosis inductors and potential anticancer drugs

The correlation models describing the quantitative structure-activity relationship have been developed for a series of the apoptosis-inducing 4-aryl-4H-chromenes and 4-aryl-2H-chromenes taking advantage of the fragment descriptors of the molecular structure. The set of models have been picked up, containing no more than 28 parameters and having the standard deviation of the predicted apoptosis-inducing activity of below 0.3 logarithmic units. Basing of the frequency of the descriptors in the 438 picked models, the structural elements crucial for the pro-apoptosis activity have been detected. The structural modifications have been suggested to enhance the studied activity of the compounds.

Highlights of This Issue

TRAIL is a potent inductor of apoptosis through activation of death receptors overexpressed on tumor cells. To improve the cytotoxic activity of TRAIL, Seifert and colleagues applied the IgE heavy-chain domain 2 (EHD2) as covalently linked homodimerization module to generate a tetravalent and bifunctional molecule by fusing an scFv to the N-terminus and a single-chain derivative of TRAIL to the C-terminus of the EHD2, combining target cell selectivity with an increased TRAIL activity. The fusion protein exhibited improved antitumor activities in vitro and in vivo and established the EHD2 as a versatile building block for the generation of targeted multivalent and multifunctional protein therapeutics.Clinically, BRAF-mutated melanoma tumors often develop acquired resistance to vemurafenib through several pathways. Effective drug combination strategies are highly desirable to overcome this resistance. Wang and colleagues discovered previously that vemurafenib-resistant cells escape G0–G1 cell-cycle arrest. Here, they demonstrated that combination of vemurafenib with a novel tubulin inhibitor, ABI-274, synergistically arrested melanoma cells in vitro and induced tumor regression in a vemurafenib-resistant xenograft model in vivo. The study offers a potential therapeutic strategy to overcome vemurafenib resistance mediated by RTK-driven PI3K pathway activation, and delay the development of vemurafenib resistance in patients.Acquired and intrinsic resistance to HER2 kinase inhibitors such as lapatinib presents a significant challenge for HER2-overexpressing breast cancers. Here, Brady and colleagues show that upregulation and/or mutation of PI3K p110α can promote acquired lapatinib resistance. The PI3K p110β isoform was dispensable for lapatinib resistance in their models. Lapatinib combined with the p110α-selective PI3K inhibitor BYL719 overcame lapatinib resistance in vitro and in animal xenografts, with minimal toxicity. These findings provide the basis for testing p110α-specific PI3K inhibition in patients to overcome lapatinib resistance and minimize the toxicity of pan-PI3K inhibitors, thus allowing maximal dosage and efficacy.Cross-talk between HER2 and IGF-IR and elevated IGF-IR signaling may contribute to resistance to trastuzumab (Herceptin; Genentech) therapy targeting HER2. Chen and colleagues developed bispecific antibody against HER2 and IGF-IR (Bi-Ab). The Bi-Ab alone or in combination with monospecific antibodies showed superior antitumor activity in vitro and in vivo compared with monospecific antibodies alone. The treatment of Bi-Ab resulted in extensive and sustainable tumor regression in SKOV-3 cancer xenografts overexpressed with HER2- and IGF-IR. This study indicates that cotargeting two or more tumor-associated cell surface receptors by bispecific antibodies may be clinically beneficial in minimizing acquired resistance to monospecific antibody therapies.

ROLE OF HEPATOPROTECTORS AND IMMUNOMODULATORS IN REGULATION OF HEPATOCYTE APOPTOSIS INDUCED BY ANTITUBERCULOSIS TREATMENT

It was currently shown that hepatopathy due to drug toxicity is associated with increased apoptosis of hepatocytes. Therefore, development of drugs which regulate cell death is of great importance. To involve some hepatoprotectors (ademethionine, reamberin, remaxol) and immunomodulators (cycloferon) into regulation of apoptosis in experimental models of liver first-line antituberculousis drugs (isoniazid, rifampicin, pyraztinamide). Levels of apoptoasis (TUNEL), expression of CD95 (receptor of tumor necrosis factor - by immunohistochemistry), expression of caspase-8, caspase-3 and pS3 (Western-blotting) were measured. Exposition offirst-line antituberculousis drugs leads to dysthrophia of liver parenchyma cells with increased apoptosis of hepatocytes and activation of CD95, caspase-8 (external way) and overexpression of p53 and caspase-3. It was found that reamberin, cycloferon and remaxol have hepatoprotective effect improving liver histology; ademethionine administered by intraperitoneal injection showed no positive effects. Reamberin demonstrated apoptosis-inhibiting effect in the experiment whereas other drugs were found to be apoptosis inductors for hepatocytes in toxic hepatopathy. Legulation of apoptosis by cycloferon and remaxol mediated by external and p53-dependent pathway is confirmed by increased expression of CD95 and p53 protein. Ademethionine might induce apoptosis by the intrinsic pathway.

Immunological Profile of Silent Brain Infarction and Lacunar Stroke

Neuroinflammation is believed to be involved in the pathophysiological mechanisms of silent brain infarcts (SBI). However, the immunological profile of SBI has been scarcely investigated. In the context of a national research project named SILENCE, aimed at investigating clinical, biochemical and pathogenic features of SBI, we have measured the plasma profile of some inflammatory-related molecules in SBI patients (n = 21), patients with recent lacunar infarcts (LI, n = 28) and healthy controls (n = 31), consecutively enrolled in four Italian centres. A panel of chemokines (MIG, CTACK, IL16, SDF1a, MCP1), growth factors (SCF, SCGFb, HGF, IL3), immunoglobulin-type adhesion molecules (ICAM1, VCAM1), proinflammatory cytokines (IL18, INFa2, MIF, IL12p40), cell surface receptors on T-cells (IL2Ra), and inductors of apoptosis (TRAIL) was assessed in plasma samples by Luminex xMAP™ technology. Immunological parameters were compared using non-parametric statistics and performance to distinguish SBI and LI was evaluated by receiver operating characteristic (ROC) analysis. Plasma levels of ICAM1 were significantly higher in both SBI and LI patients as compared to controls (SBI≥LI>Ctrl). A different trend was observed for IL16 (SBI<LI>Ctrl), SCF (LI<SBI>Ctrl) and SCGFb (SBI>LI<Ctrl). SBI subjects had significantly increased levels of MIG when compared to controls (LI≤SBI>Ctrl) and IL18 when compared to LI patients (Ctrl≤SBI>LI). All the other immunological markers did not significantly differ among groups. According to ROC analysis, the best predictor for SBI condition was the chemokine MIG (AUC = 0.84, sensitivity 86%, specificity 77%), while SCF had the best performance in distinguishing LI patients (AUC = 0.84, sensitivity 86%, specificity 68%). These results confirm the involvement of inflammatory processes in cerebrovascular disorders, particularly in SBI, a very common age-related condition. The differences in plasma profile of inflammatory molecules may underlie different pathological mechanisms in SBI and LI patients.

Abstract 5256: Effect of the CIGB-300 synthetic peptide in the modulation of signaling pathways involved with malignant progression of human and murine lung tumors.

Abstract CK2 is a serine/threonine kinase involved in cell growth, survival and apoptosis. The CIGB-300 is a synthetic peptide capable of binding to CK2 substrates thus preventing the enzyme activity. In this work we have studied the effect of IGBC-300 on several signaling pathways involved in tumor progression, cell cycle, apoptosis and biological effects associated metastatic dissemination, using a model of human and murine lung cancer cell lines (H125 and 3LL respectively). CIGB-300 presented a lethal dose 50 (LD50) of 119±2.4 μM in H125 cells and 138.3±9.9 μM in 3LL cells. Throughout an Annexin V-FITC assay we could determine that CIGB-300 is a potent apoptosis inductor since the treatment with this drug induced apoptosis at a level comparable with that induced by 10 μM of etoposide. Besides, concomitantly with cell death induction, we could observe the activation of caspase-3 and decreased expression of c-myc and cyclin D1 and D2. The levels of nuclear and cytoplasmic components of the Wnt and NFB pathways were analyzed by Western blot after treatment with CIGB-300. In the Wnt pathway, a significant decrease in nuclear β-catenin levels could be detected. Related to NFB pathway, a decrease in the nuclear p65 levels was observed, without affecting cytoplasmic levels of the IBα inhibitor. Both phenomena could be associated with a reduction in cell survival. Related to metastatic dissemination, we studied in vitro the effect of CIGB-300 on the modulation of the migratory capacity using a wound healing assay. CIGB-300 induced a significant reduction in the migratory potential in a dose dependent way in both 3LL cells (18.86 ± 2.2% vs 32.99 ± 2.7% ¼ and ½ LD50 respectively p &amp;lt;0.05) and H125 (31.73 ± 20.08% vs 51.12 ± 28.8% to ¼ and ½ LD50 respectively p &amp;lt;0.05). These results are in line with the reduction in urokinase secretion (28.64 ± 0.2% vs 44.19 ± 0.4% to ¼ and ½ LD50 respectively p &amp;lt;0.05) observed in the murine cell line 3LL treated with CIGB-300. Our results indicate that the treatment with CIGB-300 induces apoptosis and negatively modulates several signaling pathways involved in malignant progression besides affecting cell motility. This drug may become in the future a new strategy for the treatment of lung cancer, based on the blockade of different signaling pathways in lung cells. Citation Format: Alejandro J. Urtreger, Stefano M. Cirigliano, Maria I. Díaz Bessone, Carolina Flumián, Damian E. Berardi, Elisa D. Bal de Kier Joffé, Laura B. Todaro, Hernan G. Farina. Effect of the CIGB-300 synthetic peptide in the modulation of signaling pathways involved with malignant progression of human and murine lung tumors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5256. doi:10.1158/1538-7445.AM2013-5256

Quantifying Serum Level of Glycochenodeoxycholic Acid Using High Performance Liquid Chromatography in Obstructive Jaundice Patients.

Introduction. Accumulation of glycochenodeoxycholic acid (GCDC) in serum has a clinical significance as an inductor of pathological hepatocyte apoptosis, which impairs liver function. Inhibition of GCDC accumulation can be used as a marker in therapy. This study was aimed to quantify the serum level of GCDC in obstructive jaundice patients. Methodology. GCDC acid level in the serum was quantified using high performance liquid chromatography (HPLC) technique according to Muraca and Ghoos modified method. It was performed before and after decompression at day 7 and day 14. The sample was extracted with solid phase extraction (SPE) technique on SPE column. The results were analyzed using SPSS V 16.0 (P < 0.05) and quantified with standard curve on GCDC acid. Result. There were 21 cases with range of GCDC acid serum level before decompression was 90.9 (SD 205.5) μmol/L and day 7 after decompression decreased to 4.0 (SD 46.4) μmol/L and then increased to 11.3 (SD 21.9) μmol/L (P < 0.05). This method could separate GCDC acid on serum with good resolution, high precision and accuracy, and linear calibration curve on measured level range. Conclusion. HPLC can quantify GCDC acid serum on obstructive jaundice patients and can be used to support its pharmacokinetic study.

Investigation of the apoptotic way induced by digallic acid in human lymphoblastoid TK6 cells

BackgroundThe digallic acid (DGA) purified from Pistacia lentiscus. L fruits was investigated for its antiproliferative and apoptotic activities on human lymphoblastoid TK6 cells.MethodsWe attempt to characterize the apoptotic pathway activated by DGA. Apoptosis was detected by DNA fragmentation, PARP cleavage and by evaluating caspase activities.ResultsThe inhibition of lymphoblastoid cell proliferation was noted from 8.5 μg/ml of DGA. The induction of apoptosis was confirmed by DNA fragmentation and PARP cleavage. We have demonstrated that DGA induces apoptosis by activating the caspase-8 extrinsic pathway. Caspase-3 was also activated in a dose dependent manner.ConclusionIn summary, DGA exhibited an apoptosis inductor effect in TK6 cells revealing thus its potential as a cancer-preventive agent.

Current and Emerging Strategies in Bladder Cancer

Urothelial cell carcinoma is one of the most common malignancies of the urinary tract. The standard of care, intravesical chemo- and immunotherapy, while effective, is associated with a considerable side-effect profile and approximately 30% of patients either fail to respond to treatment or suffer recurrent disease within 5 years. In the setting of muscle-invasive urothelial carcinoma, use of neoadjuvant chemotherapy is associated with overall survival benefit. Muscle invasive bladder cancer is life threatening, showing modest chemosensitivity, and usually requires radical cystectomy. Although bladder cancer is fairly well-genetically characterized, clinical trials with molecularly targeted agents have, in comparison to other solid tumors, been few in number and largely unsuccessful. Hence, bladder cancer represents a considerable opportunity and challenge for alternative therapies. In this review, we will focus on promising global or pathway-based approaches (epigenetic modulators, kinase inhibitors, angiogenesis blockage, peroxisome proliferator-activated receptor γ agonists, apoptosis inductors, virus therapy) supported by a deeper understanding of molecular biology of urothelial carcinoma, which have been recently tested in clinical trials.

MicroRNA-20a inhibits stress-induced cardiomyocyte apoptosis involving its novel target Egln3/PHD3

Excessive stress, e.g. due to biomechanical overload or ischemia/reperfusion is a potent inductor of cardiomyocyte apoptosis, which contributes to maladaptive remodeling. Despite substantial progress in the understanding of the molecular pathophysiology, many components of the signaling pathways underlying remodeling in general and apoptosis in particular still remain unknown. Recent evidence suggests that microRNAs (miRs) play an important role in the heart's response to increased cardiac stress.To identify novel modulators of stress-dependent remodeling, we conducted a genome-wide miR-screen of mechanically stretched neonatal rat cardiomyocytes (NRCM). Out of 351 miRs, eight were significantly regulated by biomechanical stress, including microRNA-20a, which is part of the miR17–92 cluster. Interestingly, further expression analyses also revealed upregulation of microRNA-20a in an in vitro hypoxia/“reperfusion” model. Given the potential apoptosis-modulating properties of the miR17–92 cluster, we subjected NRCM to hypoxia and subsequent reoxygenation. AdmiR-20a significantly inhibited hypoxia-mediated apoptosis in a dose-dependent fashion, while targeted knockdown of miR-20a in NRCM induced cardiomyocyte apoptosis. Mechanistically, the antiapoptotic effect of miR-20a appears to be mediated through direct targeting and subsequent downregulation of the proapoptotic factor Egln3.Thus, miR-20a is upregulated in acute biomechanical stress as well as hypoxia and inhibits apoptosis in cardiomyocytes. These properties reveal miR-20a as a cardioprotective micro-RNA and a potential target for novel therapeutic strategies to prevent cardiac remodeling.