Upregulation Of Caspase-3 Expression Research Articles

Study on the effects of toosendanin against Eimeria tenella and its impact on the apoptosis of cecal cells in chicks

To investigate the therapeutic effect of toosendanin (TSN) against Eimeria tenella (E. tenella) in chicks. In this experiment, a chick model of artificially induced E. tenella infection was established. The anti-coccidial effect was investigated by treating different doses of TSN. A preliminary mechanism of action was conducted, using cecal cell apoptosis as a starting point. TSN at the concentration of 5 mg/kg BW showed the best effect against E. tenella with the ACI value of 164.35. In addition, TSN reduced pathological damage to cecal tissue, increased the secretion of glycogen and mucus in cecal mucosa, and enhanced the mucosal protective effect. It also elevated the levels of IFN-γ, IL-2, and IgG in serum, and raised the sIgA content in cecal tissue of infected chicks, thereby improving overall immune function. TSN was observed to promote the apoptosis of cecum tissue cells by TUNEL staining analysis. Immunohistochemistry analysis revealed that in TSN-treated groups, the expression of Caspase-3 and Bax was elevated, while the expression of Bcl-2 was reduced. TSN induced apoptosis in host cells by dose-dependently decreasing the Bcl-2/Bax ratio and upregulating Caspase-3 expression. In summary, TSN exhibited significant anticoccidial efficacy by facilitating apoptosis in host cecal cells, with the most pronounced effect observed at a dosage of 5 mg/kg body weight.

Fer-mediated activation of the Ras-MAPK signaling pathway drives the proliferation, migration, and invasion of endometrial carcinoma cells.

The role of Feline sarcoma-related protein (Fer) in various cancers has been extensively studied, but its specific involvement and underlying mechanisms in the progression of endometrial carcinoma (EC) are yet to be fully understood. The expression levels of Fer were assessed in EC tissues and cell lines using real-time quantitative PCR and western blot analysis. CCK-8 assay, Edu staining, transwell assays, and flow cytometry, were conducted to evaluate the impact of Fer on EC cells. Furthermore, a mice xenograft model and immunohistochemistry (IHC) staining were utilized for in vivo analysis. The levels of Ras, pMek1/2, and pErk1/2 were determined by western blot assay. Ras-MAPK signaling pathway inhibitor was utilized to study the regulatory role of Fer on EC cells. Our findings revealed that Fer exhibited upregulation in both EC tissues and cell lines, concomitant with the activation of the Ras-MAPK signaling pathway. Silencing of Fer resulted in the suppression of cell proliferation, migration, invasion, and Ras-MAPK signaling pathway, while promoted hypoxia-induced apoptosis in RL95-2 and KLE cells. Fer overexpression stimulated cell proliferation, migration, invasion, and Ras-MAPK signaling pathway in Ishikawa and AN3-CA cells, which were reversed after treatment with either Ras or MAPK inhibitor. Moreover, silencing of Fer suppressed tumor growth and downregulated the expression of Ki-67, Ras, pMek1/2, and pErk1/2, but had no significant effect on Mek1/2 and Erk1/2, while upregulated caspase-3 expression in vivo. In summary, the upregulation of Fer in EC cells resulted in the enhancement of cell proliferation, migration, and invasion through the activation of the Ras-MAPK signaling pathway.

Protective effect of quercetin against 5-fluorouracil-induced cardiac impairments through activating Nrf2 and inhibiting NF-κB and caspase-3 activities.

5-Fluorouracil (5-FU) is a chemotherapy used to treat many types of cancer. Cardiotoxicity is one of the common drawbacks of 5-FU therapy. Quercetin (Qu) is a bioflavonoid with striking biological activities. This research aimed to assess the ameliorative effect of Qu against 5-FU-mediated cardiotoxicity. Thirty-five rats were allocated into five groups: control group (normal saline), 5-FU group (30 mg/kg, intraperitoneally), Qu group (50 mg/kg, oral), 25 mg/kg Qu+5-FU group, and 50 mg/kg Qu+5-FU. The experimental animals were received the above-mentioned drugs for 21 days. Results showed that 5-FU significantly elevated creatine kinase, lactate dehydrogenase, serum cholesterol and triglyceride, and upregulated troponin and renin mRNA expression. Additionally, cardiac oxidant/antioxidant imbalance was evident in elevated oxidants (malondialdehyde and nitric oxide) and depleted antioxidants (superoxide dismutase, catalase, glutathione peroxidase, and glutathione). 5-FU also downregulated the gene expression of nuclear factor erythroid 2-related factor 2. Furthermore, 5-FU significantly increased cardiac pro-inflammatory cytokines (tumor necrosis factor-alpha and interleukin-1 beta) and upregulated gene expression of nuclear factor kappa-B. 5-FU significantly enhanced cardiac apoptosis through upregulating caspase-3 expression and downregulating B-cell lymphoma 2. Immunohistochemical and histopathological examinations verified the above-mentioned findings. However, all these changes were significantly ameliorated in Qu pre-administered rats. Conclusively, Qu counteracted 5-FU-mediated cardiotoxicity through potent antioxidant, anti-inflammatory, and anti-apoptotic effects.

Eupalinolide O Induces Apoptosis in Human Triple-Negative Breast Cancer Cells via Modulating ROS Generation and Akt/p38 MAPK Signaling Pathway.

Background Triple-negative breast cancer (TNBC) is a subtype of breast cancer with limited therapeutic options. Eupalinolide O (EO) was reported to inhibit tumor growth. This study is aimed at exploring the role of EO on TNBC both in vivo and in vitro. Methods. In in vitro experiments, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and clonogenic assay were conducted to measure the impact of EO on TNBC cell growth at different concentrations and time points. Flow cytometry was conducted to evaluate cell apoptosis. Mitochondrial membrane potential (MMP) loss, caspase-3 activity, and reactive oxygen species (ROS) generation were assessed. The expressions of apoptosis-related mRNAs and Akt/p38 MAPK signaling pathway-related proteins were measured. In in vivo experiments, by injecting TNBC cells into the nude mice to induce xenograft tumor, mice were treated with EO for 20 days. Then, in vivo bioluminescence imaging system was utilized to monitor the growth and distribution of TNBC cells. Tumor volume and weight were also recorded. Hematoxylin-eosin (HE) staining and ELISA assay were applied to observe tumor tissue morphology and ROS levels. Furthermore, western blotting was conducted to observe the expression of apoptosis-related proteins and Akt/p38 MAPK signaling pathway-associated proteins. Results EO inhibited the cell viability and proliferation of TNBC cells but not normal epithelial cells. Furthermore, EO induced apoptosis, decreased MMP, and elevated caspase-3 activity and ROS content in TNBC cells. Meanwhile, the expression of apoptosis-related mRNAs and Akt/p38 MAPK pathway-related proteins was regulated by EO treatment. Besides, in vivo experiments demonstrated EO not only suppressed tumor growth, Ki67 expression, ROS generation, and Akt phosphorylation but also upregulated caspase-3 expression and p-38 phosphorylation. Conclusion EO may induce cell apoptosis in TNBC via regulating ROS generation and Akt/p38 MAPK pathway, indicating EO may be a candidate drug for TNBC.

Suppressed XIAP and cIAP expressions in human brain cancer stem cells using BV6- and GDC0152-encapsulated nanoparticles

BackgroundSmac mimetics functioned against inhibitors of apoptosis proteins (IAP) often fail to achieve sufficient ability in glioblastoma multiforme (GBM) treatment due to the obstruction of the blood-brain barrier (BBB). MethodsBV6- and GDC0152-encapsulated solid lipid nanoparticles (SLNs) with surface transferrin (Tf) and folic acid (FA) (BV6-GDC0152-Tf-FA-SLNs) were developed to downregulate IAP in U87MG cells and human brain cancer stem cells (HBCSCs) for GBM treatment. Significant findingsAn increase in stearic acid (SA) level enlarged SLNs with improved entrapment of BV6 and GDC0152. 1H NMR study revealed hydrogen bonding between SA and the two IAP antagonists, supporting controlled release of BV6 and GDC0152 from SLNs. Increasing sodium dodecyl sulfate concentration reduced the size of SLNs, and raised the particle stability. Conjugated Tf and FA on SLNs favored permeating the BBB as corroborated from decreased transendothelial electrical resistance, and raised BBB permeability of propidium iodide, BV6 and GDC0152. The ability of BV6-GDC0152-Tf-FA-SLNs to target IAP in U87MG cells and HBCSCs was validated from the downregulated XIAP and cIAP-1 expressions and upregulated caspase-3 expression in immunofluorescence staining, flow cytometry and western blot analysis. Hence, Tf- and FA-grafted SLNs can be an effective colloidal delivery system to deliver BV6 and GDC0152 across the BBB, facilitate IAP targeting, and enhanced drug bioavailability of the two Smac mimetics in GBM cells for potential clinical trials.

MiR-222-3p inhibits formation of medulloblastoma stem-like cells by targeting Notch2/c-myc signaling pathway

Background Medulloblastoma (MB) is an embryonal tumor of the cerebellum, which commonly occurs in childhood. Herein, we investigated the effects of miR-222-3p on the formation of MB stem-like cells via the Notch2/c-myc pathway. Methods Quantitative real-time PCR (qRT-PCR) or western blotting was performed to determine the expression of miR-222-3p and Notch2, c-myc, proliferating cell nuclear antigen (PCNA), and caspase-3. Luciferase reporter gene, RNA immunoprecipitation (RIP), and RNA pull-down assay were applied to confirm the interaction between miR-222-3p and Notch2. Cell growth was examined by Cell Counting Kit-8. Cell cycle distribution and the number of stem cell marker CD133+ cells were examined using flow cytometry. The sphere formation assay was performed. Results miR-222-3p expression was decreased and Notch2 expression was increased in human medulloblastoma cells. miR-222-3p overexpression inhibited cell viability and the sphere formation, induced cell cycle arrest, decreased the number of CD133+ cells, and up-regulated caspase-3 expression and down-regulated PCNA, Notch2, and c-myc expression. However, Notch2 overexpression counteracted these effects of miR-222-3p overexpression. Simultaneous overexpression of Notch2 and miR-222-3p increased the c-myc promoter-luciferase activity which was decreased by miR-222-3p overexpression. Luciferase reporter gene, RIP, and RNA pull-down assay revealed that miR-222-3p targeted Notch2. Conclusion MiR-222-3p suppressed cell viability, altered cell cycle distribution, and inhibited the formation of MB stem-like cells via the Notch2/c-myc pathway.

Naringenin and morin reduces insulin resistance and endometrial hyperplasia in the rat model of polycystic ovarian syndrome through enhancement of inflammation and autophagic apoptosis.

Polycystic Ovary Syndrome (PCOS) is a gynecologic disorder with unsatisfactory treatment options. Hyperandrogenism and insulin resistance (IR) are two symptoms of PCOS. The majority of PCOS patients (approximately 50% to 70%) have IR and moderate diffuse inflammation of varying degrees. We investigated in-vitro and in-vivo effects of naringenin, morin and their combination on PCOS induced endometrial hyperplasia by interfering with the mTORC1 and mTORC2 signaling pathways. The vaginal smear test ensured the regular oestrous cycles in female rats. Serum cytokines (TNF-α and IL-6) were assessed using the ELISA test, followed by in-vivo and in-vitro determination of prominent gene expressions (mTORC1and C2, p62, LC3-II, and Caspase-3 involved in the inflammatory signaling mechanisms through RT-PCR, western bloting, or immunohistochemical analysis. In addition, the viability of naringenin or morin treated cells was determined using flow cytometry analysis. The abnormal oestrous cycle and vaginal keratosis indicated that PCOS was induced successfully. The recovery rate of the oestrous cycle with treatments was increased significantly (P<0.01) when compared to the PCOS model. Narigenin, morin, or a combination of the two drugs substantially decreased serum insulin, TNF-α, IL-6 levels with improved total anti-oxidant capacity and SOD levels (P<0.01). Treatments showed suppression of HEC-1-A cells proliferation with increased apoptosis (P<0.01) by the upregulation of Caspase-3 expression, followed by downregulation of mTORC, mTORC1, and p62 (P<0.01) expressions with improved LC3-II expressions (P<0.05) respectively. The histological findings showed a substantial increase in the thickness of granulose layers with improved corpora lutea and declined the number of cysts. Our findings noticed improved inflammatory and oxidative microenvironment of ovarian tissues in PCOS treated rats involving the autophagic and apoptotic mechanisms demonstrating synergistic in-vitro and in-vivo therapeutic effects of treatments on PCOS-induced endometrial hyperplasia.

In vitro evaluation of cytotoxic and apoptotic activities of ethanolic extract of sweet apricot kernel on PANC-1 pancreatic cancer cells

PurposeThis study aimed to evaluate the cytotoxic/apoptotic effects of sweet apricot kernel ethanolic extract (SAEE) on human cancerous PANC-1 and 293/KDR normal cells.Design/methodology/approachThe extract was prepared by maceration, and its chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS). The biological effects of SAEE on PANC-1 and 293/KDR cells were investigated using MTT (3–(4, 5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide) assay, DAPI (4',6-diamidino-2-phenylindole) and AnnexinV/propidium iodide (PI) staining. The expression of pro- and anti-apoptotic genes was evaluated by real-time quantitative polymerase chain reaction (real-time q-PCR) analysis.FindingsThe SAEE showed the selective growth inhibitory activity against PANC-1 cells with an IC50 (the 50% inhibitory concentration) value of about 1 mg/mL at 72 h. Further investigations by DAPI staining and flow cytometry revealed nucleus fragmentation and elevation of apoptotic cells, respectively. Also, a significant decrease in B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated x protein (Bax) ratio (0.41, p = 0.001) and the up-regulation of caspase-3 expression (1.5 fold, p = 0.002) indicated the induction of apoptosis in PANC-1 cells but not in 293/KDR non-cancerous cells. These results suggest that SAEE could induce apoptosis in cancer cells via a mitochondrial dependent pathway. Furthermore, GC-MS analysis showed that the SAEE is rich in γ-sitosterol and γ-tocopherol. Overall, the findings suggest that because of the selective impacts of SAEE on PANC-1 cells, it can be considered as a supportive care in adjuvant therapy for pancreatic cancer. However, the potent anticancer effects of main components of SAEE and its clinical value as an antitumor drug should be further investigated.Research limitations/implicationsConsiderable limitations of this study were that the related mechanisms of selective impacts of SAEE on cancerous and normal cells and potent cytotoxic/apoptotic effects of γ-sitosterol and γ-tocopherol as major components of SAEE were not investigated.Originality/valueRecently, a growing interest has been dedicated to plant-based natural products. Sweet apricot kernel exerts a number of pharmacological activities; however, the anticancer effect, related mechanisms and its active compounds were rarely investigated. In this study, the authors aimed to evaluate the cytotoxic/apoptotic effects of SAEE on human cancerous PANC-1 and 293/KDR normal cells.

Kruppel-like factor 4 upregulates the resistance to apoptosis induced by tumor necrosis factor α in synovial fibroblasts with rheumatoid arthritis

Objective The objective is to examine the effect of tumor necrosis factor α (TNFα) on apoptosis and proliferation of rheumatoid arthritis synovial fibroblasts (RASFs) and to elucidate the regulatory roles of Kruppel-like factor 4 (KLF4) in TNFα-induced RASF apoptosis. Methods Changes in cell proliferation were measured using an 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide (MTT) assay, and changes in cell apoptosis were detected by flow cytometry and Hoechst 33258 staining. Changes in the apoptosis-related protein caspase-3 and the apoptosis-related genes bcl-2/bax were measured by western blot and real-time PCR, respectively Results TNFα stimulation increased cell proliferation ( p &lt; 0.05), decreased cell apoptosis ( p &lt; 0.05), declined caspase-3 expression ( p &lt; 0.05), and upregulated bcl-2/bax level ( p &lt; 0.05) in RASFs. KLF4 gene silencing decreased cell proliferation ( p &lt; 0.05), increased cell apoptosis ( p &lt; 0.05), upregulated caspase-3 expression ( p &lt; 0.05), and downregulated bcl-2/bax level ( p &lt; 0.05) induced by TNFα in RASFs. Conclusions TNFα caused a decrease in RASF apoptosis, and KLF4 promoted resistance to TNFα-induced apoptosis and cell proliferation.

A novel tri-culture model for neuroinflammation.

Neuroinflammation is believed to play a primary role in the pathogenesis of most neurodegenerative diseases including Alzheimer's disease, Parkinson's disease and schizophrenia. Currently, suitable in vitro neuroinflammation models for studying cellular interactions and inflammatory mechanisms at the neurovascular unit are still scarce. In this study, we established an experimentally flexible tri-culture neuroinflammation model combining murine microglial cells (N11), mouse neuroblastoma Nuro2A cell lines and brain microvascular endothelial MVEC(B3) cells in a transwell co-culture system stimulated with lipopolysaccharides. Neuroinflammation was induced in this tri-culture model as manifested by activated N11 cells via toll-like receptor 4, resulting in increased release of proinflammatory mediators (nitric oxide, interleukin-6 and tumour necrosis factor-α) through the activation of nuclear factor-κB signalling pathway. The released inflammatory cytokines from N11 in turn, damaged the tight junction in microvascular endothelial MVEC(B3) cells, increased permeability of endothelial barrier, and induced tau phosphorylation and up-regulated caspase-3 expression in mouse neuroblastoma Nuro2A cell lines, leading to neuroinflammation injury. In summary, this tri-culture inflammation model mimics the microenvironment, the cellular crosstalk and the molecular events that take place during neuroinflammation. It provides a robust in vitro model for studying neuroinflammation mechanisms and screening for potential therapeutics to treat various neurodegenerative diseases.

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.

Nrf2 participates in the anti-apoptotic role of zinc in Type 2 diabetic nephropathy through Wnt/β-catenin signaling pathway

Zinc (Zn), as an essential trace element, has been approved to serve many roles in diabetic studies. Also Zn deficiency will aggravate renal damage in diabetes through suppression of nuclear factor-erythroid 2-related factor 2 (Nrf2) expression and function. The purpose of this study was to illustrate the role of Zn in renal apoptosis in diabetes and whether Nrf2 participated in the process. Type 2 diabetes mice model was induced by a single dose of streptozotocin (STZ) injection after high-fat diet (HFD) feeding for 3 months, then the mice were given diets supplemented with different concentrations of Zn (control, 30 ppm; low-concentration, 0.85 ppm). After 12-week treatment, morphology and associated protein expressions were examined. The results showed that low Zn diet significantly aggravated the level of renal apoptosis during diabetes, performed as the upregulation of caspase-3 expression. In addition, either low Zn diet or diabetes or both dramatically decreased the expression of Nrf2 and P-AKT in kidney. Moreover, the expression of β-catenin in kidney was increased markedly in diabetic groups. Mechanistic study applying human renal tubular epithelial cells (HK11) confirmed the role of Nrf2, as silencing Nrf2 expression abolished Zn supplementation protection against high sugar + high fat + low Zn-induced apoptosis and downregulation of β-catenin expression. All these results suggest that Nrf2 plays a key role in Zn protection against Type 2 diabetes induced renal apoptosis, which might be through Wnt/β-catenin signaling pathway.

Inhibitory effects of tanshinone IIA from Salvia miltiorrhiza Bge on human bladder cancer BIU-87 cells and xenograft in nude mice

Abstract The objective of this study was to investigate the inhibitory effects of tanshinone IIA on human bladder cancer BIU-87 cells and the xenograft in nude mice. BIU-87 cells were treated with tanshinone IIA with concentration of 0.5, 1, 2 and 4 g/L. The cell proliferation, cycle and apoptosis were detected. The nude mice with BIU-87 cell xenograft were treated with normal saline (control), 5-fluorouracil (5-FU), 200, 400 and 600 mg/kg tanshinone IIA, respectively. The animal body weight and growth of tumor were measured. The expressions of B-cell lymphoma-2 (Bcl-2), Caspase-3 and proliferating cell nuclear antigen (PCNA) protein in xenograft were detected. The apoptosis index of cells was determined. Results showed that, tanshinone IIA inhibited the proliferation of BIU-87 cells, promoted their apoptosis, and arrested more cells in G0/G1 phase. Tanshinone IIA significantly inhibited the growth of xenograft in nude mice and promoted the apoptosis of tumor cells in xenograft. Tanshinone IIA down-regulated Bcl-2 and PCNA expression and up-regulated Caspase-3 expression in xenograft. In conclusion, tanshinone IIA has inhibitory effects on human bladder cancer BIU-87 cells, and can inhibit the BIU-87 cells xenograft in nude mice.

Amelioration of Monosodium Glutamate-induced Testicular Damage and Infertility in Male Rats by Water Melon and Cantaloupe Seeds Extract and Juices

Aims: Monosodium glutamate (MSG) is extensively used as food additive and flavor enhancer, there is a growing concern that this may affect the male reproductive system and fertility. The objective of this study is to investigate the effect of MSG on fertility and testes of mature male rats and the ameliorative role of water melon and cantaloupe (seeds extract and juices).&#x0D; Study Design: Thirty-six male Sprague - Dawely rats (150-180 g) were randomly assigned into six groups (n=6). Group (1): orally administered with distilled water. Group (2): orally administered with 60 mg/kg of MSG. Groups (3 and 4): orally administered with 60 mg/kg of MSG + 200 mg/kg of water melon seeds extract and juice respectively. Groups (5 and 6): orally administered with 60 mg/kg of MSG + 200 mg/kg of cantaloupe seeds extract and juice respectively.&#x0D; Results: Results showed that administration of MSG for 6 weeks caused abnormalities of semen characteristics, increased DNA damage and up-regulation of caspase3 expression in the testes tissue. Also, the levels of plasma sex hormones were decreased and the oxidant-antioxidant status was disturbed, moreover, MSG caused alteration in the histopathological structures of testicular tissue. Administration of seeds extract or juices of water melon and cantaloupe almost corrected the biochemical and histopathological alteration produced by MSG.&#x0D; Conclusion: This study concluded that water melon and cantaloupe seeds and juice extracts have an ameliorative role against MSG-induced testicular damage and infertility in rats.

Anti-cancer effects of chemotherapeutic agent; 17-AAG, in combined with gold nanoparticles and irradiation in human colorectal cancer cells.

The present study evaluated the anti-cancer effects of irradiation (Ir) alone, Ir after heat shock protein 90 inhibitor; 17-allylamino-17-demethoxygeldanamycin (17-AAG) and gold nanoparticle (GNP) treatments in human colorectal cancer cell line (HCT-116), with the targeting of related mechanisms. Water-soluble tetrazolium salt-1 assay was utilized to study the cytotoxic effects of 17-AAG, GNP, Ir in single and combination cases on the cell viability of HCT-116 cells. The cells were examined with DNA fragmentation electrophoresis and evaluated for apoptosis induction. Caspase-3 expression as a critical apoptosis element in protein level was detected by western blotting. Treatment with 17-AAG in a dose dependent manner for 24h inhibited the cellular viability of HCT-116 cells. GNP at a dose of 70μM had the lowest cytotoxic effects and was thus selected for combination treatment studies. Based on the results, GNP at a dose of 70μM did not have a significant effect on cellular viability of HCT-116. In contrast, the evaluation of double and triple combinations, GNP with Ir (2Gy of 6 MV X-ray radiation) and 17-AAG in double combinations induced significant cytotoxicity. Both DNA damage pattern and caspase-3 protein upregulation were present in Ir,GNP/17-AAG,GNP and Ir,17-AAG combinations compared to single treatments. Furthermore, in the three combination of GNP,Ir,17-AAG, radiosensitization effects (increased caspase-3 expression) occurred with a minimum concentration of 17-AAG. According to the results of this study, 17-AAG as chemotherapeutic agent in combination with Ir and GNP exerts noticeable anti-cancer effects, inhibited cell viability, and increased apoptosis occurrence by upregulating caspase-3 expression. It is suggested that these combinations should be more evaluated as a promising candidate for colorectal cancer treatment. Graphical abstract Anti-cancer effects of chemotherapeutic agent; 17-AAG, in combined with gold nanoparticles and irradiation in human colorectal cancer cells.

Biological activities and chemical components of Potentilla kleiniana Wight & Arn

In this study, we investigated the antimicrobial, antioxidant, and cytoprotective activities of ethanol extract and the ethyl acetate (EtOAc) fraction of P. kleiniana Wight & Arn. The EtOAc fraction exhibited antimicrobial effects against most of the microorganisms that were tested, including Staphylococcus aureus, Candida albicans, Pseudomonas aeruginosa, and, Escherichia coli, but not Aspergillus niger. In addition to its excellent antioxidant activity, the EtOAc fraction attenuated the UVB-induced cell death via upregulation of caspase-3 expression in human keratinocytes. The HPLC/ESI-MS/MS analysis allowed identification of the components in the EtOAc fraction. Overall, our results suggest that P. kleiniana is a valuable source of bioactive compounds for the development of pharmaceuticals and cosmetics.

Evaluation of Iodine-125 Interstitial Brachytherapy Using Micro-Positron Emission Tomography/Computed Tomography with 18F-Fluorodeoxyglucose in Hepatocellular Carcinoma HepG2 Xenografts

BackgroundIodine-125 interstitial brachytherapy (125I-IBT) is a promising treatment option for unresectable hepatocellular carcinoma (HCC). This study evaluated the usefulness of micro-positron emission tomography/computed tomography (micro-PET/CT) with 18F-fluorodeoxyglucose (18F-FDG) in assessing response to 125I-IBT in HCC HepG2 xenograft.Material/MethodsTwelve mice with bilateral HepG2 xenografts were divided into 3 equal groups implanted with iodine-125 seeds into the left xenografts with a dose of 30, 50, and 80 Gy, respectively, and the right xenografts were used as internal controls. Before and 28 days after treatment, the 18F-FDG micro-PET/CT was performed. The ratios of left to right xenografts of tumor volume (RTV), maximum standardized uptake value (RSUVmax), mean optical density of caspase-3 expression (RMODcaspase-3), and apoptosis index (RAI) were compared.ResultsThe RTV means of the 50 and 80 Gy groups were significantly lower than in the 30 Gy group after treatment (P<0.01) and the RTV means after treatment were lower than baseline in the 50 and 80 Gy groups (P<0.05). The RSUVmax mean after treatment was lower than baseline in the 80 Gy group (P<0.05). The RMODCaspase-3 and RAI means of the 80 Gy group were higher than in the 30 Gy group (P<0.05). The RSUVmax was correlated negatively to RMODcaspase-3 (r=−0.624, P<0.05) and RAI (r=−0.651, P<0.05).ConclusionsThis study suggest that 125I-IBT inhibits tumor growth via upregulating caspase-3 expression and prompting apoptosis in HCC HepG2 xenografts. The 18F-FDG micro-PET/CT may be a useful functional imaging modality to assess early response to 125I-IBT in HCC HepG2 xenograft.

Ethylacetate extract from Tetrastigma hemsleyanum inhibits proliferation and induces apoptosis in HepG2 and SMMC-7721 cells.

PurposeThis study aimed to investigate the effect of ethylacetate extract from Tetrastigma hemsleyanum (EET) on the proliferation and apoptosis of HepG2 and SMMC-7721 cells and determine the underlying mechanisms.Materials and methodsHepG2 and SMMC-7721 cells were cultured in vitro until the exponential growth phase and then treated with different concentrations of EET for 24 h. We performed a colony forming assay to determine colony forming ability, CCK8 assay to detect cell proliferation, Annexin V–FITC/PI double staining to analyze cell apoptosis, and Western blot to measure the protein expression of Caspase-3, Bcl-2, and Bax.ResultsEET significantly inhibited the proliferation of HepG2 and SMMC-7721 cells in a concentration- and time-dependent manner (P<0.05). After treatment with 0, 50, 100, 150, 200, and 250 μg/mL EET for 24 h, HepG2 the proliferation rates were 100.00%±0.00%, 90.33%±1.76%, 67.67%±0.88%, 47.33%±0.88%, 37.00%±0.00%, and 30.33%±0.67%, respectively, and 100.00%±0.00%, 18.25%±1.05%, 19.99%±0.59%, 23.42%±0.46%, 29.70%±0.79%, and 29.8%±0.41% for SMMC-7721 cells, respectively. After treatment with 0, 50, 100, 150, 200, and 250 μg/mL EET for 24 h, the apoptotic rates were 11.08%±0.72%, 27.44%±0.51%, 32.92%±0.41%, 26.20%±0.47%, 22.92%±0.24%, and 55.60%±0.08%, for HepG2 cells, respectively, and 59.18%±0.17%, 41.24%±2.05%, 52.54%±0.39%, 50.54%±1.08%, and 57.44%±1.93% for SMMC-7721 cells, respectively. Compared with the treatment groups, the control group showed a significantly lower apoptotic rate (47.91%±1.09%, P<0.05). EET at the different concentrations downregulated the protein expression of Caspase-3 in HepG2 cells and upregulated it in SMMC-7721 cells; it also downregulated the protein expression of Bcl-2 in HepG2 and SMMC-7721 cells and upregulated the protein expression of Bax in HepG2 and SMMC-7721 cells.ConclusionThese findings suggest that EET exerts antiproliferative and proapoptotic effects against HepG2 and SMMC-7721 cells mediated by downregulation or upregulation of Caspase-3 expression. Our study may help to develop EET for the pharmacological treatment of hepatoblastoma or hepatocellular carcinoma.

TRIM52 plays an oncogenic role in ovarian cancer associated with NF-kB pathway

Emerging evidence suggests that the members of the tripartite motif (TRIM) family play a crucial role in cancer development and progression. The purpose of the study was to explore TRIM52's role in tumorigenesis and its potential molecular mechanism in ovarian cancer. The study demonstrated that knockdown of TRIM52 in SKOV3 and CAOV3 cells inhibited ovarian cancer cell invasion, migration, and proliferation, and induced cell apoptosis. On the contrary, overexpression of TRIM52 in HO8910 cells showed contrary results. Further, overexpression of TRIM52 enhanced the expression of phosphorylated IKKβ and IKBα proteins and nuclear protein P65, which implied the activation of NF-kB signal pathway. Knockdown of TRIM52 downregulated the mRNA and protein levels of NF-kB signal downstream effectors of the NF-kB pathway, including MMP9, Bcl2, IL8, and TNFα, but upregulated caspase-3 expression. These results suggested that activation of the NF-kB pathway is involved in TRIM52-mediated regulation in ovarian cancer. The nude mice study further confirmed that knockdown of TRIM52 blocked tumor growth, inhibited cell proliferation, and promoted cell apoptosis. Our data strongly suggested that TRIM52 plays an oncogenic role in ovarian cancer development associated with the NF-kB signal pathway and may be a potential target for cancer therapy.

Aflatoxin B1 impairs spermatogenesis: An experimental study for crosslink between oxidative stress and mitochondria-dependent apoptosis.

The present experimental study was carried out to investigate the crosslink between aflatoxin B1 (AFB1)-induced oxidative stress and mitochondria-dependent apoptosis in testicles. For this purpose, 24 mature male Swiss albino mice were randomly divided into control and test groups. The AFB1 was dissolved in corn oil and ethanol (95:5, v/v) vehicle. The animals in test group subdivided into three groups, which received the AFB1 at a daily dose of 20 μg/kg body weight, through intraperitoneal (i.p.) route, for 7, 14, and 21 days. The mice in the control group received the vehicle alone for 21 days. The expression of Bcl-2, Bax, p53, and caspase-3 at both mRNA and protein levels were analyzed by using reverse transcription PCR (RT-PCR) and immunohistochemistry, respectively. Moreover, the mitochondrial content of germinal epithelium, tubular differentiation (TDI), and spermiogenesis (SPI) indices was analyzed. Finally, the apoptosis was assessed by using TUNEL staining. Observations revealed that the AFB1 remarkably (P < .05) reduced Bcl-2 expression at both mRNA and protein levels. Up-regulated Bax, caspase-3, and p53 expression were revealed in AFB1-received animals, which developed time-dependently. Histological examinations exhibited a significant reduction in TDI and SPI indices. Finally, the AFB1-induced apoptosis index increased time-dependently. In conclusion, the AFB1 adversely affects the spermatogenesis via inducing oxidative stress, diminishing cellular mitochondrial content and enhancing pro-apoptotic Bax, caspase-3, and p53 expression. All these impairments result in mitochondria-dependent apoptosis.