Down-regulation Of Bcl-2 Expression Research Articles

Investigation of biological activity of nickel (II) complex with naproxen and 1,10-phenanthroline ligands

After the accidental discovery of cis-platinum, extensive attempts have centralized on the rational design of metallic compounds for cancer treatment. Here a solvent-dependent complex of nickel (II) with 1,10-phenanthroline and naproxen, [Ni(1,10-phenanthroline)(naproxen)2(solvent)], solvent = 83% H2O and 17% EtOH in the crystal structure, has been synthesized and specified by the X-ray structure analysis. It’s in vitro DNA binding was inspected by the multispectroscopic methods and gel electrophoresis. The data of DNA-viscosity and competition fluorimetric test by methylene blue (MB) and Hoechst 33258 confirm groove binding mode of the complex to CT-DNA. Comparison of the results of this binding study with previous work revealed that the mode of binding of small compounds to DNA is highly influenced by the structure of the compounds. The DNA cleavage potency of the complex was appraised by the agarose gel electrophoretic and it was found that the complex does not have any momentous cleavage potency on the pUC18 plasmid DNA. The cytotoxicity of the complex on HT 29, HepG2 and HEK-293 cell lines by MTT method indicates that %inhibition of the complex on HT 29 is better than HepG2, compared with cisplatin drug. On HEK-293 cells, %inhibition growth of normal cells of the complex is less than cisplatin. Flow cytometry analysis of the complex on the HT 29 cells indicated the apoptosis cell death. RT-PCR studies revealed down-regulation of BCL2 expression, while the expression of BAX, caspase 3 and BAX/BCL2 genes was up-regulated in HT 29 cells by the complex. Highlights A solvent-dependent nickel (II) with naproxen and 1,10-phenanthroline with aqueous solubility was synthesized and characterized. All experimental results indicate a groove mode of binding of the complex to CT-DNA. Potential biological characteristics confirmed that the complex is a promising candidate as anticancer agent. Communicated by Ramaswamy H. Sarma

Biological and Molecular Evidences for the Antitumor and Antioxidant Potential of Quercetin In Vitro

To investigate the protective mechanisms of quercetin in promoting HeLa cell apoptosis and protecting human umbilical vein endothelial cells (HUVECs) from oxidative damage, in this study, MTT test was preformed to observe the inhibition rate of quercetin on HeLa cell proliferation (50.0, 75.0 and 100.0 mol/L), and the apoptosis rate was measured with flow cytometry. Western blotting was used to detect the protein expression levels in signaling pathways in quercetin-treated HeLa cells and HUVECs. MTT assay results showed that quercetin could inhibit (P < 0.001) the proliferation of HeLa cells in a time-concentration dependent manner. Apoptosis results showed that quercetin can increase the early and late apoptosis rate (P < 0.001). Western blotting revealed that quercetin can down-regulate BCL-2 expression level (P < 0.05), up-regulate BAX and CASPASE-3 expression level (P < 0.05) in HeLa cells. Flow detection results showed that the number of HUVECs labeled with CD105/CD62E and CD31/CD34 in the oxidation group decreased significantly (P < 0.01), increased significantly (P < 0.001) in the drug group. Quercetin can decrease the expression levels of p16, p53 and p21 significantly (P < 0.05). In conclusion, quercetin can promote the apoptosis of HeLa cell by regulating the expression level of CASPASE-3, BCL-2 and BAX; and protect HUVECs against oxidative damage and inhibit the apoptosis by down-regulating p16, p53 and p21.

Regulatory T cells are less sensitive to glucocorticoid hormone induced apoptosis than CD4+ T cells

Earlier we have reported that thymic regulatory T cells (Treg) are resistant to in vivo glucocorticoid hormone (GC)-induced apoptosis, while the most GC-sensitive DP thymocytes died through the activation of mitochondrial apoptotic pathway. Here we analyzed the apoptosis-inducing effect of high dose (10–6 M) in vitro dexamethasone (DX) treatment in mouse thymic- and splenic Tregs and CD4+ T cells. Activation of both extrinsic and intrinsic apoptotic pathways started after 2 h of DX treatment in CD4 SP thymocytes and was 3 × higher than in CD4+ splenocytes, while in Treg cells, weak activation of the extrinsic apoptotic pathway started only after 3 h. We also investigated the expression of 21 apoptosis-related molecules using a protein array and found higher level of both pro-and anti-apoptotic molecules in Tregs compared to CD4+ T cells. 4 h in vitro DX treatment induced upregulation of most apoptosis-related molecules both in Tregs and CD4+ T cells, except for the decrease of Bcl-2 expression in CD4+ T cells. We found high basal cytosolic Ca2+ levels in untreated Treg cells, which further increased after DX treatment, while the specific TCR-induced Ca2+ signal was lower in Tregs than in CD4+ T cells. Our results suggest that in the background of the relative apoptosis resistance of Treg cells to GCs might be their high basal cytosolic Ca2+ level and upregulated Bcl-2 expression. In contrast, downregulation of Bcl-2 expression in CD4+ T cells can explain their higher, DX-induced apoptosis sensitivity.

Genotoxic and Cytotoxic Activities of Lantadene A-Loaded Gold Nanoparticles (LA-AuNPS) in MCF-7 Cell Line: An in vitro Assessment.

Gold nanoparticles (AuNPs) have been widely used in many applications. Their usage as drug delivery vehicles has also gained considerable attention due to their chemical and optical properties as well as their good biocompatibility. The present study was conducted to evaluate the efficiency of AuNPs in enhancing the cytotoxic and apoptotic induction activity of lantadene A (LA), separated from Lantana camara leaves, on the breast tumor cell line MCF-7 in vitro. By utilizing plant-mediated synthesis method of nanostructures, LA-loaded AuNPs (LA-AuNPs) were prepared and their formation was confirmed by means of ultraviolet-visible spectroscope, atomic force microscope, scanning electron microscope, and zeta potential. The cytotoxic effect of LA-AuNPs was analyzed using a methylthiazol tetrazolium assay and compared to free AuNPs and LA. The results indicated a significant increase in the reduction of MCF-7 cells viability after incubation with LA-AuNPs. As determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, LA-AuNPs induced a greater ratio of DNA-fragmented cells compared to LA-treated and untreated cells. Also, by operating real-time polymerase chain reaction, LA-AuNPs-treated cells displayed an increased upregulation of p53 expression and downregulation of BCL-2 expression in addition to a significant reduction in the level of BCL-2-BAX ratio. No significant effect was shown on the expression of BAX. Collectively, our results indicate that LA-AuNPs showed promising cytotoxicity to MCF-7 cells as a novel nanoscale preparation, likely via induction of apoptotic genes and stimulation of DNA fragmentation.

RETRACTED ARTICLE: Two Mixed-Ligand Co(II)-Based Coordination Polymers: Treatment Activity on Pediatric Asthma Via Inducing Eosinophil Apoptosis Through Down-Regulating Bcl-2 Expression

In this work, two coordination polymers containing Co(II) ions as nodes, namely, [Co(NDC)(L)(H2O)]n (1, H2NDC = 1,4-naphthalene dicarboxylic acid) and [Co(DCTP)(L)·(H2O)2]n (2, H2DCTP = 2,5-dichloroterephthalic acid) were prepared based on 1,4-bis(5,6-dimethylbenzimidazole)butane (L), the flexible N-donor ligand and different dicarboxylate ligand. Their treatment activity on the pediatric asthma was evaluated in the biological research. Firstly, the enzyme linked immunosorbent assay (ELISA) detection was carried out to determine the IL-1β and TNF-α content in the bronchus. The inflammatory response in the bronchial epithelial cells was determined by the real time reverse transcription-polymerase chain reaction (RT-PCR). After that, the eosinophil apoptosis in the airway was measured. Finally, the western was performed for the confirmation of eosinophil apoptosis by detecting Bcl-2 and caspase-3.

New manganese(II), iron(II), cobalt(II), nickel(II) and copper(II) saccharinate complexes of 2,6-bis(2-benzimidazolyl)pyridine as potential anticancer agents

New mononuclear complexes [Mn(NO3)(sac)(H2O)(bzimpy)]·2DMF (Mn), [Fe(sac)2(H2O)(bzimpy)]·2H2O (Fe), [Co(bzimpy)2](sac)2·2H2O (Co), [Ni(bzimpy)2](sac)2·H2O·i-PrOH (Ni) and [Cu(sac)2(bzimpy)]·3DMF (Cu) (sac = saccharinate and bzimpy = 2,6-bis(2-benzimidazolyl)pyridine) were synthesized and structurally characterized by elemental analysis, UV–Vis, IR, ESI-MS and X-ray diffraction. The anticancer activity of the metal complexes against A549 (lung), MCF-7 (breast), HT29 (colon) cancer cells and MCF10A (normal human breast epithelial) cells was tested and compared with those of cisplatin and bzimpy. The complexes displayed potent cytotoxic activity especially in MCF-7 and A549 cell lines, but they were practically inactive against the normal cells. Mechanistic studies with Mn and Cu complexes on A549 cells indicated that the complexes induced G0/G1 arrest. Both complexes increased intracellular ROS (reactive oxygen species) levels and successfully caused both mitochondrial dysfunction and double-strand DNA breaks. The up-regulated Bax and down-regulated Bcl-2 expression levels, caspase-3/7 activation and reduced Fas expression indicated that Mn and Cu induced ROS-dependent mitochondria-mediated intrinsic apoptosis in A549 cells.

MiR-1184 regulates the proliferation and apoptosis of colon cancer cells via targeting CSNK2A1

MicroRNA (miRNA) exerts an important part in colon cancer cell proliferation and apoptosis. Meanwhile, the dysregulation of some miRNAs is detected in colon cancer cells. However, it remains unclear about the underlying mechanism of their effects on tumor pathogenesis. The current work aimed to examine the miR-1184 effect on colon cancer cells. The differentially expressed miRNAs (DEMs), including miR-9-3p, miR-1184, miR-492, miR-92a-1-5p and miR-20a-3p, were obtained from the GSE115108 and GSE132619 data sets using the ‘GEO2R’ online tool. Based on the findings, miR-1184 was significantly down-regulated within colon cancer cells and tissues. Moreover, the experimental results of CCK8, flow cytometry, colony formation and Western blotting assays showed that, miR-1184 over-expression suppressed colon cancer cell proliferation through inhibiting Ki67 expression and promoted their apoptosis through up-regulating cleaved caspase-3 and down-regulating Bcl-2 expression. By contrast, miR-1184 inhibition exerted the opposite effects. A total of 110 target genes of miR-1184 were predicted using the TargetScan and miRTarBase databases, which were then used to construct the protein-protein interaction (PPI) network based on the DAVID and STRING websites and to perform GO and KEGG pathway enrichment analyses. The MCODE plug-in of cytoscape was utilized to verify that CSNK2A1 was the target gene and key gene in significant modules. MiR-1184 directly targets CSNK2A1 via using RNA immunoprecipitation assay and luciferase reporter gene assay. According to the results, CSNK2A1 over-expression reversed the functions of miR‐1184 over-expression in suppressing colon cancer cell proliferation and enhancing their apoptosis. In conclusion, over-expression of miR-1184 inhibits colon cancer cell proliferation but promotes their apoptosis through down-regulating CSNK2A1 expression.

Synthesis and anticancer activity of some pyrimidine derivatives with aryl urea moieties as apoptosis-inducing agents

A new series of pyrimidine derivatives containing aryl urea moieties was designed and synthesized. The anticancer activities of all compounds were evaluated in vitro against colon and prostat cancer cell lines by MTT assay. Among these compounds, 4b exhibited the highest cytotoxic activity against SW480 cancer cell line with IC50 value of 11.08 µM. Mechanistic studies showed that compound 4b arrested cell cycle at G2/M phase and induced apoptosis through upregulating Bax, Ikb-α and cleaved PARP and downregulating Bcl-2 expression levels. Moreover, compound 4b induced loss of mitochondrial membrane potential in SW480 cells. These results suggest that pyrimidine with urea moieties could be a template for designing new anticancer agents.

Prunus Armeniaca L. Seed Extract and Its Amygdalin Containing Fraction Induced Mitochondrial-Mediated Apoptosis and Autophagy in Liver Carcinogenesis.

Despite significant advances in therapeutic interventions, liver cancer is the leading cause of cancer mortality in the world. Potential phytochemicals have shown to be promising agents against many life-threatening diseases because of their low toxicity and potential effectiveness. The current study aims to conduct an in vitro investigation of the anticancer activity of Apricot Extract (AE) and Amygdalin Containing Fraction (ACF), additionally studying their therapeutic effects on DMBAinduced liver carcinogenesis mice model to highlight their related biochemical and molecular mechanisms. Amygdalin was isolated from the seeds of P. armeniaca L. Male mice received AE or ACF, DMBA, DMBA+AE, DMBA+ACF, and vehicles. The oxidative stress and antioxidant markers, cell proliferation by flow cytometric analysis of Proliferating Cell Nuclear Antigen (PCNA) expression, angiogenesis marker (VEGF), inflammatory marker (TNF-α), apoptotic, anti-apoptotic and autophagy genes expression (caspase-3, Bcl-2, and Beclin-1) were investigated. AE and ACF were found to stimulate the apoptotic process by up-regulating caspase-3 expression and down-regulating Bcl-2 expression. They also reduced VEGF and PCNA levels and increased the antioxidant defense system. Moreover, AE and ACF treatments also inhibited HepG2 and EAC cell proliferation and up-regulated Beclin-1 expression. This study provides evidence that, in DMBA-induced hepatocarcinogenesis, the key proteins involved in the proliferation, angiogenesis, autophagy, and apoptosis are feasible molecular targets for hepatotherapeutic potential using AE and ACF.

A protective role of resveratrol against the effects of immobilization stress in corpora lutea of mice in early pregnancy

In the present study, we aimed to investigate a protective role for resveratrol against the effects of immobilization stress on corpora lutea (CL) of mice in early pregnancy. A total of 45 early-pregnant mice were divided into no immobilization stress (NIS) group, immobilization stress (IS) group, and immobilization and resveratrol treatment (IS+RES) group (n=15). Mice were immobilized in plastic tubes (50 mL) for 3 h per day during day 1 to 7 of pregnancy. In the IS+RES group, 5 mg kg−1 d−1 of resveratrol was administered just prior to application of stress. We analyzed apoptotic activity in CL by Western blotting analysis (WB), transmission electron microscopy (TEM), and immunohistochemistry (IHC). Serum progesterone levels were examined with radioimmunoassay (RIA). IHC results showed that the intensity of positive staining for Bax was increased, and for Bcl-2 was decreased in CL after IS, while resveratrol treatment reversed the positive staining for Bax and Bcl-2. WB revealed that immobilization stress up-regulated the expression of Bax and caspase-9, and down-regulated Bcl-2 expression, while resveratrol treatment attenuated the effects of immobilization stress on the expression of Bax, Bcl-2 and caspase-9. According to our TEM results, apoptosis as defined by chromatin condensation was found in CL after immobilization stress, while resveratrol inhibited the apoptosis. We also demonstrated that immobilization stress decreased progesterone concentrations and ovarian expression of StAR, while resveratrol restored the concentrations of progesterone and expression of StAR back to normal. These results indicated that immobilization stress induced luteal regression while resveratrol inhibited luteal regression, suggesting that resveratrol plays a protective role on corpora lutea of mice during early pregnancy.

Imidazole-thiazolidinone inhibits oesophageal cancer cell proliferation via induction of apoptosis and cell cycle arrest at S phase

Purpose: To investigate the effect of imidazole-thiazolidinone on oesophageal cancer (OC) cell proliferation, and the mechanism of action involved.Methods: Human OC cells (HCE-6 and KYSE-1170) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin solution at 37 ˚C for 24 h in a humidified atmosphere of 5 % CO2 and 95 % air. After attaining 60 - 70 % confluency, the cells were treated with serum-free medium and graded concentrations of imidazolethiazolidinone (up to 160 μM) for 24 h. Normal cell culture without imidazole-thiazolidinone served as control. Cells in logarithmic growth phase were selected and used in this study. Cell proliferation and apoptosis were assessed using 3 (4,5 dimethyl thiazol 2 yl) 2,5 diphenyl 2H tetrazolium bromide (MTT), and flow cytometric assays, respectively. The levels of expression of apoptosis-related proteins were determined using Western blotting.Results: Treatment of HCE-6 and KYSE-1170 cells with imidazole-thiazolidinone for 48 h led to significant and dose-dependent reduction in their proliferation, as well as significant and dosedependent increase in the number of apoptotic cells (p < 0.05). Light microscopy revealed significantreduction in HCE-6 cell count, detached cells, reduced cell size and irregular cytoplasmic vacuoles. Imidazole-thiazolidinone treatment significantly and dose-dependently decreased HCE-6 and KYSE-1170 cell migration, and arrested HCE-6 cell cycle at S phase (p < 0.05). In HCE-6 cells, imidazolethiazolidinone treatment significantly and dose-dependently upregulated the expressions of cleaved caspase-3/8/9 and bax, but down-regulated bcl-2 expression significantly and dose-dependently (p < 0.05). However, metalloproteinases 2 and 9 (MMP-2 and MMP-9) expressions in HCE-6 and KYSE-1170 cells were significantly and dose-dependently down-regulated by imidazole-thiazolidinone treatment (p < 0.05).Conclusion: The results obtained in this study suggest that imidazole-thiazolidinone suppresses OC cell proliferation via induction of apoptosis and arrest of cell cycle at S phase.
 Keywords: Imidazole-thiazolidinone, Oesophageal cancer, Metastasis, Cell cycle arrest, Apoptosis

Citral Induced Apoptosis through Modulation of Key Genes Involved in Fatty Acid Biosynthesis in Human Prostate Cancer Cells: In Silico and In Vitro Study

The isomers of citral (cis-citral and trans-citral) were isolated from the Cymbopogon citratus (DC.) Stapf oil demonstrates many therapeutic properties including anticancer properties. However, the effects of citral on suppressing human prostate cancer and its underlying molecular mechanism have yet to be elucidated. The citral was isolated from lemongrass oil using various spectroscopic analyses, such as electron ionized mass spectrometry (EI-MS) and nuclear magnetic resonance (NMR) spectroscopy respectively. We carried out 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to evaluate the cell viability of citral in prostate cancer cells (PC-3 and PC3M). Furthermore, to confirm that PC3 undergoes apoptosis by inhibiting lipogenesis, we used several detection methods including flow cytometry, DNA fragmentation, Hoechst staining, PI staining, oil staining, qPCR, and Western blotting. Citral impaired the clonogenic property of the cancer cells and altered the morphology of cancer cells. Molecular interaction studies and the PASS biological program predicted that citral isomers tend to interact with proteins involved in lipogenesis and the apoptosis pathway. Furthermore, citral suppressed lipogenesis of prostate cancer cells through the activation of AMPK phosphorylation and downregulation of fatty acid synthase (FASN), acetyl coA carboxylase (ACC), 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR), and sterol regulatory element-binding protein (SREBP1) and apoptosis of PC3 cells by upregulating BAX and downregulating Bcl-2 expression. In addition, in silico studies such as ADMET predicted that citral can be used as a safe potent drug for the treatment of prostate cancer. Our results indicate that citral may serve as a potential candidate against human prostate cancer and warrants in vivo studies.

Progress of the anoikis effector bcl-2 inhibitor of transcription 1

bcl-2 inhibitor of transcription 1 (Bit1) is an anoikis effector that can play a role of caspase-independent apoptosis by down-regulating bcl-2 expression. Previous studies have found that Bit1 affects cell survival and apoptosis through Erk and FAK-PI3K-Akt-NF-κB pathways. It is worth noting that the expression of Bit1 has shown the obvious tumor specificity in different tumors, and it is closely related to TNM stage, differentiation and prognosis of the tumor. This review summarizes the expression, main mechanism, and significance of Bit1 in different tumors. Key words: Apoptosis; Neoplasms; Bcl-2 inhibitor of transcription 1

Epigallocatechin-3-gallate protects Wharton's jelly derived mesenchymal stem cells against in vitro heat stress

The deteriorating effects of heat stress abrogate the therapeutic implications of human Wharton's jelly derived mesenchymal stem cells (hWJMSCs) transplanted in burn wounds. Topically applied green tea extract comprising epigallocatechin-3-gallate (EGCG) is known to repair burn wounds. Here, we investigated the protective role of EGCG priming of hWJMSCs against heat-induced stress in vitro along with the involved underlying mechanism. EGCG ameliorated heat-induced injuries as demonstrated by significantly improved cell morphology, viability, triggered cell migration and enhanced expression of heat shock proteins. In addition, decreased lactate dehydrogenase release and reduced percentage of senescent and apoptotic cells were observed. EGCG priming alleviated the detrimental effects of thermal stress in hWJMSCs as observed by significant down-regulation in expression of BCL2 associated X (BAX), interleukin 6 (IL6), and interleukin 1 beta (IL1β) genes, while proliferating cell nuclear antigen (PCNA), BCL2 like 1 (BCL2L1), vascular endothelial growth factor (VEGF), transforming growth factor beta 1 (TGFβ1), hepatocyte growth factor (HGF) and interleukin 4 (IL4) genes were up-regulated. Accompanying gene expression data, EGCG primed cells exposed to heat stress also exhibited remarkably increased secretion of VEGF, HGF, epidermal growth factor (EGF), stromal-derived factor 1 (SDF1) proteins while the reduced release of IL-6, and tumor necrosis factor-alpha (TNF-α) proteins. Further, synergistic activation of extracellular-signal-regulated kinase (ERK) and protein kinase B (PKB/AKT) proteins was observed. These findings suggest that EGCG priming might enhance the therapeutic efficacy of hWJMSCs in the burnt tissue through regulation of ERK and AKT signaling pathways, and improved cellular responses.

Trisenox Disrupts MDM2-DAXX-HAUSP Complex and Induces Apoptosis in a Mouse Model of Acute Leukemia.

Trisenox (TX) is successfully used for both de novo and relapsed acute promyelocytic leukemia (APL) treatment. Although TX toxicity to APL cells is mediated by oxidative stress, DNA damage, cell cycle arrest, and apoptosis, its mode of action in the transgenic mice model of APL is poorly understood. We hypothesized that TX regulates cell cycle and apoptosis in APL mice by p53 activation, DNA damage, and reduced expression of MDM2-DAXX-HAUSP complex. To test hypothesis, we treated APL mice with different doses (0, 1.25.2.5.5.0 & 7.5 mg/kg body wt) of TX and collected the liver and bone marrow cells. We applied several techniques to check the expression of PML-RARα, complex molecules, and DNA damage in APL mice bone marrow cells and liver. Our findings indicate that TX reduced the expression of PML-RARα and complex molecules, induced DNA damage and activated p53 leading to cell cycle arrest and apoptosis in APL mice liver. We found that TX promoted more promyelocytes formation with dense granules in bone marrow cells. It also transmitted the DNA damage signal through protein kinase (ATM & ATR) leading to disruption of complex and activation of p53 in APL mice liver. TX induced cell cycle arrest through activation of p53, p21, and reduced expression of cyclin D1 and cyclin dependent kinases (CDK 2, 4 & 6) in mice liver. It also caused apoptosis through upregulation of caspase 3 and Bax expression, and down-regulation of Bcl2 expression. Taken together, these molecular targets provide new insights into TX mode of action in APL mice.

MicroRNA-16 Down-regulates BCL2 Expression and Induces Papillary Thyroid Carcinoma Cell Apoptosis Via Extracellular-regulated Kinase Pathway

MicroRNA-16 Down-regulates BCL2 Expression and Induces Papillary Thyroid Carcinoma Cell Apoptosis Via Extracellular-regulated Kinase Pathway

Protective Role of Hydrogen Gas on Oxidative Damage and Apoptosis in Intestinal Porcine Epithelial Cells (IPEC-J2) Induced by Deoxynivalenol: A Preliminary Study.

To explore the protective role of hydrogen gas (H2) on oxidative damage and apoptosis in intestinal porcine epithelial cells (IPEC-J2) induced by deoxynivalenol (DON), cells were assigned to four treatment groups, including control, 5 μM DON, H2-saturated medium, and 5 μM DON + H2-saturated medium treatments. After 12 h of different treatments, the cell viability, biomarkers of cell redox states, and gene expression of antioxidant enzymes and apoptosis were observed and detected. Furthermore, caspase-3 and Bax protein expressions were measured by Western blot analysis. Our results demonstrated that the 5 μM DON significantly caused cytotoxicity to IPEC-J2 cells by reducing cell viability and increasing lactate dehydrogenase release in culture supernatants. Moreover, DON treatments significantly increased levels of 8-hydroxy-2′-deoxyguanosine, 3-nitrotyrosine, and malonaldehyde; however, they decreased total superoxide dismutase and catalase activities and downregulated messenger RNA (mRNA) expression related to antioxidant enzymes in cells. The 5 μM DON treatment also downregulated Bcl-2 expression and upregulated caspase-3 and Bax expression. However, the H2-saturated medium significantly improved cell growth status and reversed the change of redox states and expression of genes and proteins related to apoptosis induced by DON in IPEC-J2 cells. In conclusion, H2 could protect IPEC-J2 cells from DON-induced oxidative damage and apoptosis in vitro.

Role of protein kinase D1 in regulating the growth, apoptosis and drug sensitivity of oral squamous carcinoma cells

This study aimed to investigate the role of protein kinase D (PKD)1 in regulating the growth, apop-tosis, and drug sensitivity of the squamous carcinoma cell line SCC-25. The SCC-25 cell line was transfected with either the control-shRNA or PKD1-shRNA plasmids. The stable transfected cells were selected, and the efficiency of PKD1 knockdown was detected by Western blot. The growth and apoptosis of SCC-25 were analyzed with a cell counting kit-8 (CCK8) and flow cytometry. The 50% inhibitory concentrations (IC50) of paclitaxel in the control and PKD1 knockdown cell lines were detected by CCK-8. The expression levels of Bax, Bcl-2, and P-gp were detected by Western blot. PKD1 was constitutively expressed and phosphorylated in various cancer cell lines. Inhibiting the expression of PKD1 in SCC-25 cells by RNA interference could inhibit the growth and promote the apoptosis of SCC-25 cells via downregulating Bcl-2 expression. Additionally, inhibiting PKD1 expression could downregulate the expression of P-gp, thereby decreasing both the IC50 and resistance index of paclitaxel. PKD1 plays an important role in regulating the biobehavior of SCC-25. It is a potential therapeutic target for oral squamous carcinoma.

M-phase phosphoprotein 8 promotes gastric cancer growth and metastasis via p53/Bcl-2 and EMT-related signaling pathways.

The main issue of this study is to demonstrate whether M-phase phosphoprotein 8 (MPP8) affect gastric tumor growth and metastasis. Retrospective study was proceeded in 280 patients' surgical specimens with different disease stages. Loss-of-function assays, including 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide, flow cytometry, and transwell assays were performed to evaluate the biological function of MPP8 in gastric cancer cells. Apoptosis and metastasis relative biomarkers were measured by quantitative real-time polymerase chain reaction and Western blot analysis. Compared with normal adjacent tissues, obviously elevated MPP8 expression was found in gastric cancer tissues. Elevated MPP8 expression was associated with male sex (vs female sex), intermediate differentiation (vs poorly differentiated cancer), and later stage (vs earlier stage). Furthermore, MPP8 overexpression in tumor tissues was marginally associated with a poor prognosis, with a significant relationship between MPP8 overexpression and prognosis among patients with poorly differentiated gastric cancer. Inhibition of MPP8 in these cells significantly suppressed proliferation and colony formation, promoted apoptosis, and repressed invasion. Furthermore, silencing of MPP8 remarkably increased apoptosis-related proteins (p53, Bax, and PARP) expression, but downregulated Bcl-2 expression. Silencing of MPP8 also decreased the expression of metastasis pathway-related proteins (N-cadherin and vimentin), and as well as the levels of anti-oncogene ZEB1, MET, and KRAS mRNA. Our findings demonstrated that MPP8 might be an oncogene by positively regulating gastric cancer cell function through the p53/Bcl-2 and epithelial to mesenchymal transition-related signaling pathways.

Effect of hexavalent chromium exposure on the liver and kidney tissues related to the expression of CYP450 and GST genes of Oreochromis niloticus fish: Role of curcumin supplemented diet

The present study evaluated the adverse effects of the hexavalent chromium (Cr (VI)) at sub-lethal concentrations and the ameliorative potential of curcumin (CUR) over a sub-chronic exposure period on Oreochromis niloticus. Fish were exposed to Cr (VI) (4.57 mg/L) and CUR (0.02% in diet or 200 mg/kg diet), individually or in combination for 60-days. The growth rate during the period of experiment, condition factor, body composition, hepatosomatic index (HSI), hematological parameters, oxidative stress, apoptotic and DNA damage, branchial, hepato- and nephrotoxicity were estimated in this study. Moreover, the changes in mRNA expression of Cytochromes (CYP450) and glutathione S-transferase (GST) in kidney and liver tissues were assessed by qRT-PCR. Additionally, the concentration of metallothionine in the liver, histological investigation, and lesion scoring to the branchial, hepatic, renal and gill tissues were applied. The results revealed that Cr (VI) exposure caused a significant decline in most hematological variables and growth rate with down-regulation of CYP450 and GST expression. Histologically, Cr (VI) induced diverse forms of cell injury, vascular, and inflammatory alterations with upregulation of caspase-3 and downregulation of Bcl2 expression in the examined tissues. Additionally, it elevated the levels of serum MDA and 8-hydroxy-2' -deoxyguanosine than control. CUR-supplementation resulted in a significant improvement in most indices, amelioration of histological alterations and up-regulation of CYP450 and GST expression. These results may conclude that dietary supplements with CUR could be useful for modulation of the growth with protective effects to the branchial, hepatic, and renal tissues in response to Cr (VI) exposure, thereby presenting a promising feed additive for Nile tilapia in aquaculture.