Hoechst 33342 Research Articles

Mesalazine induces apoptosis via mitochondrial pathway in K562 cell line.

Inflammation is an initial response of the body to infection and relationship between inflammation and cancer has been established. Nuclear factor kappa B (NF-κB) is a central factor in inflammation and its activity contributes to tumor progression and apoptosis prevention consequently leading to cancer promotion. As a result, NF-κB inhibitors can cause apoptosis. In this study, the effect of mesalazine as a NF-κB inhibitor on growth and apoptosis of K562 cells has been investigated. The K562 cells were first cultured in RPMI-1640 medium containing 10.00% fetal bovine serum. After that, they were treated for 72 hr with different concentrations of mesalazine (20.00, 40.00, 60.00 and 80.00 μM mL-1). The MTT assay was used to evaluate cell viability. Hoechst staining and RT-PCR of apoptosis related genes (Bcl-2 and Bax) were carried out to illustrate apoptosis induction and immunocytochemistry was performed to investigate changes in c-Myc protein level. According to the results of MTT assay, all of applied mesalazine concentrations decreased K562 cells viability. Hoechst staining showed that the fragmented nuclei increased indicating apoptosis induction. Immuno-cytochemical ‎ results showed that mesalazine decreased c-Myc in treated cells. The RT-PCR results also showed an increase in Bax and a decrease in Bcl-2 expressions in mesalazine-treated cells. As the results suggest, mesalazine reduces cell viability by inducing apoptosis in K562 cell line; therefore, it can be used as a candidate for the leukemia treatment.

The Apoptosis of Liver Cancer Cells Promoted by Curcumin/TPP-CZL Nanomicelles With Mitochondrial Targeting Function.

The mitochondrion is one of the most important cellular organelles, and many drugs work by acting on mitochondria. Curcumin (Cur)-induced apoptosis of HepG2 in liver cancer cells is closely related to the function of inhibiting mitochondria. However, the mitochondrion-targeting curcumin delivery system was rarely been reported. It is important to develop a high-efficiency mitochondrion-targeting curcumin vector that can deliver curcumin into mitochondria directly. Here, a special mitochondrion-targeting delivery system based on triphenylphosphine bromide (TPP)-chitosan-g-poly-(N-3-carbobenzyloxy-l-lysine) (CZL) with TPP functional on the surface is designed to perform highly efficient mitochondria-targeting delivery for effective liver cancer cell killing in vitro. The TEM images showed that the nanomicelles were spherical; the results of fluorescence test showed that TPP-CZL nanomicelles could promote the cellular uptake of drugs and finally targeted to the mitochondria. The results of cell survival rate and Hoechst staining showed that curcumin/TPP-CZL nanomicelles could promote the apoptosis of liver cancer cells. Curcumin/TPP-CZL nanomicelles could significantly reduce the mitochondrial membrane potential, increase the expression of pro apoptotic protein Bcl-2, and reduce the expression of antiapoptotic Bax protein, and these results were significantly better than curcumin/CZL nanomicelles and curcumin. It is a potential drug delivery system with high efficiency to target mitochondria of liver cancer cells.

New organoruthenium(II) complexes containing N, X-donor (X = O, S) heterocyclic chelators: Synthesis, spectral characterization, in vitro cytotoxicity and apoptosis investigation

Four Schiff base ligands namely ((1E)-1-(1-(6-chloro-2-oxo-2H-chromen-3-yl) ethylidene)semicarbazide) (HL1), ((1E)-1-(1-(6-chloro-2-oxo-2H-chromen-3-yl)ethylidene thiosemicarbazide) (HL2), ((1E)-1-(1-(7-hydroxy-2-oxo-2H-chromen-3-yl)ethylidene) semicarbazide) (HL3) and ((1E)-1-(1-(7-hydroxy-2-oxo-2H-chromen-3-yl)ethylidene) thiosemicarbazide) (HL4) have been prepared, and the direct reaction of these ligands with [RuCl2(η6-p-cymene)]2 yielded the new cationic ruthenium(II) arene complexes (C I - C IV). The compounds were characterized using various spectroscopic methods. Spectroscopic analysis revealed that the ligands coordinated to ruthenium ion in a neutral bidentate fashion via N & S/O atoms. The in vitro anticancer activity of the ligands and complexes have been screened using MTT assay with two cancer cell lines, namely MCF-7 (human breast cancer cells) and A549 (human lung carcinoma). Interestingly, the cytotoxic nature of the complexes is much more potent than the standard drug cisplatin and their parent ligands with low IC50 values. The non-toxic nature of the compounds has been confirmed with human umbilical vein endothelial cells HUVEC cells. The morphological changes of the MCF-7 and A549 cells with the complexes C I- C IV have been studied by AO-EB (Acridine Orange-Ethidium Bromide) and Hoechst staining methods, and these results revealed that the complexes induced cell death only through apoptosis. The cytotoxic nature of the compounds was found to be C IV > C III > C II > C I > cisplatin > ligands. Complex C IV showed better activity due to the electron-donating hydroxyl group present in the 7th position of the coumarin ring. These results highlighted the strong possibility to develop highly active ruthenium arene complexes as anticancer agents.

Localised light delivery on melanoma cells using optical microneedles.

Light-based therapy is an emerging treatment for skin cancer, which has received increased attention due to its drug-free and non-invasive approach. However, the limitation of current light therapy methods is the inability for light to penetrate the skin and reach deep lesions. As such, we have developed a polylactic acid (PLA) microneedles array as a novel light transmission platform to perform in vitro evaluation regarding the effect of light therapy on skin cancer. For the first time, we designed and fabricated a microneedle array system with a height fixation device that can be installed in a cell culture dish and an LED array for blue light irradiation. The effect of the blue light combined with the microneedles on cell apoptosis was evaluated using B16F10 melanoma cells and analyzed by Hoechst staining. Our results demonstrate that blue light can be transmitted by microneedles to skin cells and effectively affect cell viability.

Apoptosis inducing potential of Cyclosporine C in human lung cancer cells (A549)

Cyclotides are a large group of plant based cyclic peptides that have attracted attention in the field of peptide drug design. Cyclic peptides exhibit a more potent biological activity when compared to linear peptides due to their stable cyclic structure. Cyclosporine C is a naturally occurring lipophilic cyclic peptide which is used as an immunosuppressant drug. The objective of this study was to establish Cyclosporine C as a viable novel anticancer drug in the treatment of lung cancer and to find the optimal concentration at which Cyclosporine C results in a significant decrease of viable cancer cells to determine the physiological changes caused by Cyclosporine C and also to find the pathway of apoptosis that is induced by Cyclosporine C. The cytotoxic effect of Cyclosporine C was assessed using techniques such as MTT, Hoechst staining, DNA fragmentation, Flowcytometry and Western blot which were carried out on Lung adenocarcinoma cell line A549. The results obtained showed a significant decrease in A549 viable cells at a concentration of 200μg/mL of Cyclosporine C as observed by the MTT assay. The nuclear condensation and DNA fragmentation of apoptotic cells were observed using Hoechst 33258 staining and DNA fragmentation assays respectively. Western blot confirms that Cyclosporine C treated cells expressed a significant level of caspase-9 enzyme. From the results obtained, we can conclude that Cyclosporine C induces apoptosis in Lung adenocarcinoma cell line A549. The apoptosis is induced by caspace-9 enzyme dependent pathway. The antitumor activity of Cyclosporine C suggests that it has potential as a novel anticancer drug for human lung cancer treatment.

Histone deacetylase HDAC2 regulates microRNA-125a expression in neuroblastoma.

BackgroundNeuroblastoma (NB) is an infrequent childhood malignancy of the peripheral sympathetic nervous system and is accountable for about 10% of pediatric tumors. microRNA (miR)‐125a has been implicated to serve as a tumor suppressor in various cancers. Herein, we set out to ascertain whether miR‐125a exerts antitumor effects in NB.MethodsDownregulated miRNAs were identified by miRNA microarray analysis of NB tissues and paracancerous tissues. The expression of miR‐125a in NB tissues and cells was detected by reverse transcription‐quantitative (RT‐q) PCR, followed by prognostic analysis. Gene Ontology (GO) enrichment analysis was performed on target genes of differentially expressed miRNAs. Cell proliferation, apoptosis, and differentiation were detected by cell counting kit‐8 (CCK‐8), Hoechst staining, immunofluorescence, and western blot. NB cells were injected into nude mice to detect tumorigenic, apoptotic, and differentiation activities in vivo. Dual‐luciferase assay and chromatin immunoprecipitation (ChIP) were carried out to verify the binding relationship between miR‐125a and PHOX2B or histone deacetylases 2 (HDAC2), respectively. Finally, rescue experiments were conducted.ResultsmiR‐125a was downregulated in NB tissues and cells, which was associated with poor prognosis. miR‐125a reduced NB cell proliferation and augmented apoptosis and differentiation. NB cells with miR‐125a overexpression decreased cell tumorigenesis and increased apoptosis and differentiation in xenograft tumor tissues. miR‐125a targeted PHOX2B, which was highly expressed in NB tissues and cells. HDAC2, highly expressed in NB tissues and cells, repressed miR‐125a transcription through histone deacetylation. Overexpression of HDAC2 or PHOX2B rescued the effects of miR‐125a on NB cell proliferation, apoptosis, and differentiation.ConclusionHDAC2 inhibited miR‐125a transcription through deacetylation, and miR‐125a suppressed NB development through binding to PHOX2B.

Synthesis of a novel nanobioconjugate for targeted photodynamic therapy of colon cancer enhanced with cannabidiol.

Photodynamic therapy (PDT) is a promising primary treatment option for colorectal cancer (CRC), however CRC is accelerated by resilient CRC stem-like cells, which decrease its efficacy. In recent years, researchers have shown an emerging interest in the anticancer stem cell effects of cannabidiol (CBD). This study developed a targeted nanobioconjugate for specific ZnPcS4 photosensitizer intracellular accumulation within in vitro cultured human CRC cells (CaCo-2) for enhanced PDT primary treatment, as well as limited its secondary spread by combining this treatment with CBD. The final nanobioconjugate (FNBC) was successfully synthesized and characterized using various methods. The cytotoxicity of the FNBC and CBD were tested on CRC cells using laser irradiation at 673 nm with a fluency of 10 J/cm2. 24 h post treatment, morphological changes were assessed via microscopy, cell viability was measured using Annexin V-FITC and cellular nuclear DNA was visualized under fluorescent microscopy, following Hoechst staining. FNBC and CBD combinative treatment induced the most significant photodamage, leaving a staggering 6%*** viable cells. Overall, through active targeting of CRC cells using the FNBC, the enhanced PDT primary treatment of CRC was achieved, and the combinative treatment with CBD noted significant limitations on its secondary spread.

In vivo and in vitro inhibition of SCLC by combining dual cancer-specific recombinant adenovirus with Etoposide.

Oncolytic virotherapy is emerging as an important modality in cancer treatment. In a previous study, we designed and constructed Ad-Apoptin-hTERTp-E1a (Ad-VT), a dual cancer-selective anti-tumor recombinant adenovirus. To explore the therapeutic effect of recombinant adenovirus Ad-VT together with Etoposide on small cell lung cancer, the ability of Ad-VT alone, Etoposide alone, and a combination of Ad-VT + Etoposide to inhibit proliferation of NCI-H446 and BEAS-2B cells was investigated using the WST-1 method. According to the inhibitory action of different combinations, a combination index (CI) was estimated by CalcuSyn software to select the best combination. The inhibitory effect of Ad-VT combined with Etoposide on NCI-H446 and BEAS-2B cells was detected by crystal violet staining and the CFST method. Hoechst, Annexin V and JC-1 staining were used to explore the inhibitory pathway of Ad-VT combined with Etoposide on NCI-H446 cells. The migratory and invasive abilities of treated NCI-H446 cells were assessed by Transwell and BioCat methods. Tumor volume, body weight and survival rate were measured to analyze the anti-tumor and toxic effects of different treatments in tumor-bearing mice. Ad-VT (20 MOI) combined with Etoposide (400nM) significantly inhibited NCI-H446 cell proliferation with reduced toxicity of Etoposide to normal cells. Ad-VT induced apoptosis of NCI-H446 cells mainly through the mitochondrial apoptosis pathway, an effect significantly increased by the combined treatment. Ad-VT together with Etoposide significantly inhibited migration and invasion of NCI-H446 cells, inhibited tumor growth in vivo and prolonged the survival of tumor-bearing mice. The above results indicate that when combined with Etoposide, Ad-VT may have an important role in synergistically inhibiting tumors.

Effect of 1,4-Napthoquinone Derivatives on Anti-proliferation and Apoptosis Induction in Skin and Lung Cancer Cells

Background: Cancer is a major public health concern around the world. Nowadays, standard therapy in combination with targeted therapy are being used for cancer treatment, but these therapeutics show side effects or unwanted symptoms and high cost. 1,4-Napthoquinone and their derivatives were used in cancer research for a long time with high efficiency than some cancer drug. In the present study, the effect of a novel 1,4-napthoquinone, DL667 and DL666 on apoptosis induction in malignant melanoma, A375 cells and lung cancer, A549 cells, respectively were studied. Objective: To investigate the effects of 1,4-napthoquinone derivatives, DL667 and DL66, on apoptosis induction in malignant melanoma, A375 and lung cancer, A549 cells, respectively. Materials and Methods: Cell growth inhibition was determined by MTT assay. Apoptosis induction was via chromatin condensation or apoptotic bodies determined by Hoechst staining. In addition, changes in mitochondrial membrane potential (ΔΨm) were done by JC-1 staining. Results: The results showed that DL667 inhibited cell growth in A375 with an IC50 at 11.2+0.419 μM in a concentration-dependent manner and DL666 inhibited cell growth of A549 cells at 200 μM. Hoechst staining assay revealed that both DL667 and DL666 could induce chromatin condensation and apoptotic bodies in A375 and A549 cells. In addition, the JC-1 staining showed that DL667 and DL666 decreased the ΔΨm, a marker event of early apoptosis via mitochondria, in A375 and A549 treated cells. Conclusion: DL667 and DL666 showed anti-proliferation and apoptosis induction in A375 and A549 cells, respectively. Moreover, DL667 and DL666 induced chromatin condensation, apoptotic bodies and decreased the ΔΨm, resulting in apoptosis cell death induction. Our results suggested that the decrease of ΔΨm may be further developed as a model for study in other cancer cell lines. However, the underlying mechanisms of apoptosis induction associated with signaling should be further studied. Keywords: 1,4-Napthoquinone; Apoptosis; Chromatin condensation; Apoptotic bodies; Mitochondrial membrane potential

Trigger of apoptosis in adenocarcinoma gastric cell line (AGS) by a complex of thiosemicarbazone and copper nanoparticles

Seeking novel anticancer agents with minimal side effects against gastric cancer is vitally important. Copper, as an important trace element, takes roles in different physiologic pathways. Also, there is a higher demand for copper in cancer cells than normal ones. Copper complexes containing a therapeutic ligand could be promising candidates for gastric cancer chemotherapy. In this work, copper oxide nanoparticles were synthesized, functionalized with glutamic acid (CuO@Glu) and conjugated with thiosemicarbazone (CuO@Glu/TSC NPs). The NPs were characterized and their antiproliferative potential against AGS cancer cells was investigated using MTT, flow cytometry, Hoechst staining, and caspase 3 activation assays. The FT-IR results showed the proper binding of TSC to CuO@Glu NPs and crystallinity of the prepared NPs was confirmed by the XRD pattern. The EDX analysis confirmed the presence of Cu, N, C, O, and S elements and lack of impurities. The Hydrodynamic size and zeta potential of the CuO@Glu/TSC NPs were 168nm and 27.5mV, respectively. The NPs had spherical shape and were in a size range of 10 to 60nm in diameter. This work revealed that CuO@Glu/TSC NPs efficiently inhibited the proliferation of AGS cells with significantly lower IC50 value (203µg/mL) than normal HEK293 cells (IC50 = 435µg/mL). Flow cytometry and Hoechst staining obviously revealed apoptosis induction among CuO@Glu/TSC treated cells, and caspase-3 activity significantly increased by 1.4 folds. This study introduced CuO@Glu/TSC as an efficient anticancer against gastric cancer cells with lower toxicity toward normal cells which could be employed for cancer treatment after further studies.

VP7: In Vitro Comparison of Dynamic Seeding Versus Microinjection of Mesenchymal Stem Cells for Acellular Nerve Allograft

INTRODUCTION: Mesenchymal stem cell (MSC) supplemented acellular nerve allografts (ANA) are a potential strategy to improve treatment of segmental nerve defects. Prior to clinical translation optimal cell delivery methods must be defined. While two techniques, dynamic seeding and microinjection have been described, the seeding efficiency, cell viability, and distribution of MSCs in ANAs of the techniques have yet to be compared. MATERIALS AND METHODS: Sciatic nerve segments of Sprague-Dawley rats were decellularized and MSCs were harvested from the adipose tissue of Lewis rats. Cell viability was evaluated after injection of MSCs through a 27-gauge needle at different flow rates (10, 5, and 1 μL/min). MSCs were dynamically seeded or longitudinally injected in ANAs. Cell viability, seeding efficiency, and distribution were evaluated using Live/Dead and MTS assays, scanning electron microscopy, and Hoechst staining. RESULTS: No statistically significant difference in cell viability after injection at different flow rates was seen. After cell delivery, 84.07% ± 3.68 and 87.75% ± 2.07 of MSCs remained viable in the dynamic seeding and microinjection group, respectively (p=0.41). The seeding efficiency of microinjection (100.4%±5.6) was significantly higher than dynamic seeding (48.1%±8.6) on day 1 (p=0.001). Dynamic seeding demonstrated a significantly more uniform cell distribution over the course of the ANA compared to microinjection (p= 0.02). CONCLUSION: MSCs remain viable after both dynamic seeding and microinjection in ANAs. Higher seeding efficiency was observed with microinjection, but dynamic seeding resulted in more uniform distribution. In vivo studies are required to assess the effect on gene expression profiles and functional motor outcomes.

Ranunculus Japonicus Induces Cell Cycle Arrest, Apoptosis, Autophagy and Targeting the Akt/Mapks Pathways in Human Neuroblastoma SH-SY5Y Cells

Objective: Ranunculus japonicus thumb. has been used as a traditional medicine for the treatment of asthma, malaria, tuberculosis, hepatitis, jaundice and so on. The aim of this study was to investigate the anti-cancer effect and action mechanism of the petroleum ether extract of Ranunculus japonicus Thumb. (RJTPEE) on human neuroblastoma SH-SY5Y cells. Methods: Cell viability was detected using MTT assay. FCM and Hoechst33258 staining was used to examine the cell cycle as well as apoptosis, and in addition, Western blot was also used to examine the expression of a range of related proteins. Results: MTT assay showed that RJTPEE could inhibit SH-SY5Y cells proliferation in concentration- and time-dependent manners. FCM analysis showed that RJTPEE could induce S and G2/M phase arrest, cell apoptosis, cell autophagy, the mitochondrial membrane potential loss (ΔΨm) and increase the production of intracellular ROS of SH-SY5Y cells. Topical morphological changes of apoptotic body formation were also observed by Hoechst 33258 staining. Western blot analysis indicated that the expressions of cell cycle-related protein Cyclin B1/D1 and CDK1/2/4/6 were obviously decreased. Meanwhile, obvious increase of apoptosis-related protein expressions, such as Bax, cytochrome c, caspase-3, caspase-9, PARP-1, and decrease of Bcl-2 protein expression were observed. Furthermore, Akt/MAPKs pathway-related proteins were also detected. Moreover, the decreased expression of autophagy-related proteins LC3 I protein indicated that RJTPEE could also enhance the autophagy ability of cells. Conclusion: RJTPEE could exert the antitumor activity on SH-SY5Y cells by inducing cell cycle arrest, cell autophagy, the mitochondrion-dependent cell apoptosis and targeting the Akt/MAPKs signaling pathway. RJTPEE has a promising therapeutic potential for neuroblastoma.

Cardamonin Induces Apoptosis and Suppresses the Proliferation of BxPC3 Cells

Pancreatic cancer is one of the most lethal solid malignancies featured by rapid disease progression with no significant clinical symptoms and the treatment of pancreatic cancer remains unmet clinical need. Thus, to find a novel and potent therapeutic strategy for the treatment of pancreatic cancer is highly desired. The present study investigated the role and molecular mechanisms of cardamonin in pancreatic cancer cell line BxPC3. Cells are treated with cardamonin for 48 h and 72 h and evaluated cytotoxicity of cardamonin by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Clonogenic assay was applied to observe the anti-proliferation of cardamonin in pancreatic cancer cell line BxPC3 cells. Hoechst staining and flow cytometry were used to verify the mechanism of anti-proliferation of cardamonin in pancreatic cancer cell line BxPC3 cells. The expression of apoptosis-related proteins was evaluated by western blotting to investigate the molecular mechanism of cardamonin in pancreatic cancer cell line BxPC3 cells. The results of the present study showed that cardamonin inhibited the proliferation of pancreatic cancer cell line BxPC3 cells with the half maximal inhibitory concentration value of 28.86 μM and 22.83 μM at 48 h and 72 h respectively. Moreover, cardamonin induced apoptosis in BxPC3 cells via deoxyribonucleic acid damage to inhibit the pancreatic cancer. Overall, these results suggested that cardamonin has the potential to be used as a new therapeutic approach for pancreatic cancer.

Glutathione ameliorates Hypoxia/Reoxygenation (H/R) induced hepatocyte oxidative damage via regulating HO-1 signaling

In our study, CCK-8 assay and LDH release detection were performed to detect the optimal protective concentration of GSH on HL7702 cell viability during H/R injury. HL7702 cells were randomly divided into four groups: Control group, H/R group, H/R+GSH group, and H/R+GSH+HO-1-siRNA group. Then, reactive oxygen species (ROS) was evaluated by DHE staning, MDA, T-SOD measurements; Cell injury was detected by CCK-8, LDH release, and supernatant AST and ALT levels; Apoptosis was determined by Hoechst staining and caspase 3 level. Compared with controls, H/R caused significant HL7702 cell injury evidenced as reduced cell viability, increased LDH release and apoptotic cell death (P < 0.01), with concomitant increases in ROS and MDA production (P < 0.01), while treated with GSH in H/R group significantly attenuated oxidative injury, enhanced cell viability and downregulated cell apoptosis (P < 0.01) together with HO-1 upregulation (P < 0.01). Knockdown HO-1 by its siRNA cancelled the protective effects of GSH from H/R compared with GSH group (P < 0.01). HO-1 was induced in HL7702 exposed to H/R injury and its level was obviously overexpressed after H/R injury with GSH treatment, suggesting its protective potential in GSH against H/R injury. GSH increases the expression of HO-1, which enhanced the early antioxidative activity and played a protective role against HL7702 cells H/R injury.

Ginkgolide A Participates in LPS-Induced PMVEC Injury by Regulating miR-224 and Inhibiting p21 in a Targeted Manner.

Most studies have focused on the protective effects of ginkgolide A against ischemia/reperfusion-induced cardiomyopathy and injury of the brain, liver, and other organs, but there are few reports about the protection of lung tissues. This study was designed to clarify the protection of ginkgolide A against lipopolysaccharide (LPS)-induced pulmonary microvascular endothelial cell (PMVEC) injury. PMVECs were extracted and fell into control, LPS, and ginkgolide A groups. Next, we delved into the growth activity and apoptosis rate of cells via the CCK-8 assay and Hoechst staining, independently. Beyond that, western blotting (WB) was implemented for measurement of the expressions of cyclin D1, cyclin-dependent kinase 4 (CDK4), and CDK inhibitor (p21) that pertained to the cell cycle. The target sites of ginkgolide A were confirmed by miRNA array and real-time quantitative PCR. The relationship between miR-224 and p21 was analyzed using dual-luciferase reporter gene assay. Compared with the control group, the LPS group and ginkgolide A group had significantly decreased cell growth activity and relative expressions of cyclin D1 and CDK4 and elevated apoptosis rate and p21 expression. Pronounced elevations were observable in the cell growth activity and expressions of cyclin D1, CDK4, and p21, while the ginkgolide A group presented with a reduced apoptosis rate in comparison with the LPS group (P < 0.05). MiR-224 was the target of ginkgolide A, which had targeted regulatory effects on p21. Ginkgolide A can modulate miR-224 expression and regulate p21 expression in a targeted manner to enhance the resistance of PMVECs to LPS-induced cell apoptosis.

Monte Carlo simulation for assessing absorbed dose effects of low-dose β-radiation (90Sr/90Y) on cytotoxicity and apoptotic death in K562 cells.

Most studies suggest that dose-specific radiation regimens are essential for optimal induction of cancer cell response. This study focused on determining β-radiation-absorbed dose (rad) effects on the cell viability, cytotoxicity, hypersensitivity, and cell death of K562 cells using experimental methods and Monte Carlo simulation (MCS). The K562 cells were cultured and irradiated with β-particles emitted from a strontium source in vitro, with the estimated daily activity of 1.238 μCi. The treated cells were radiated at least three times every day for 3 consecutive days. The cell viability and apoptosis were investigated in treated cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, DNA electrophoresis, Hoechst dye, and inverted microscope. The average absorbed doses were obtained by MCS (MCNPX code). To verify simulation and experimental results, we used a Geiger-Muller counter and estimated a scaling factor. The cytotoxic effects and cell death were induced in the treated groups via rad in a time-dependent manner. The highest apoptotic and cytotoxic effects were observed in cells after irradiation with β-particles for 120 min per day in 3 consecutive days. rads were determined using MCNPX code and cell survival rates were significantly reduced during irradiation periods. No significant hyper-radiosensitivity was found based on experimental and theoretical results. Despite the difficult calculation of the rad in the target cells and the scant information in this field, fortunately we have achieved significant theoretical data consistent with the experimental results. Our findings also introduced MCS as a better choice for evaluating of rad effects under different cellular conditions with high accuracy.

Propofol induces the apoptosis of neural stem cells via microRNA-9-5p / chemokine CXC receptor 4 signaling pathway

ABSTRACT Recent studies suggested that propofol, one of the most widely used anesthetics, may cause neurotoxicity in the developing brain, leading to cognitive deficits in adults. However, the underlying mechanisms remain unclear. In this study, we aimed to evaluate the mechanisms of propofol neurotoxicity in the neural stem cells (NSCs). The mRNA and protein expression levels of microRNA-9-5p (miR-9-5p) and chemokine CXC receptor 4 (CXCR4) were determined by quantitative reverse transcription-polymerase chain reaction and Western blotting analyses. Cell viability and apoptosis were evaluated using the cell counting kit-8 and Hoechst staining kits. The levels of apoptosis-related proteins B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein, and caspase-3 were detected by Western blotting analysis. These results confirmed that propofol activated cell apoptosis in a dose-dependent manner. A significant increase in the miR-9-5p and CXCR4 expression was observed in the propofol-treated cells. The overexpression of miR-9-5p induced apoptosis in NSCs, accompanied by elevated apoptosis-related protein activity. Furthermore, mitigated CXCR4 expression reduced propofol-induced cell apoptosis. We conclude that propofol induces cell death in NSCs, and overexpression of miR-9-5p/CXCR4 contributes to propofol-induced cell apoptosis, which might be a target for developing novel strategies to treat propofol neurotoxicity.

Gossypin Suppresses Cell Growth by Cytotoxic Effect and Induces Apoptosis in MCF-7 Cells

Aim: Today, breast cancer is a disease that is encountered commonly in women with limited options for treatment. It is needed to find new agents that can be effective in preventing or managing this disease. It has been demonstrated that gossypin inhibits tumor growth. In our study, it has been targeted to examine the effects of gossypin regarding both anticancer activity and apoptosis in MCF-7 cells. Material and Method: MCF-7 cells were treated with different doses of gossypin and with 50 µM cisplatin for 24, 48, and 72 hours. The MTT analysis, Caspase-3, Caspase-9, and Nf-KB mRNA expressions of those MCF-7 cells which were treated with gossypin were also conducted in order to evaluate the apoptosis or necroptosis-induced cell death. Results: In MTT experiments, it has been observed that the administration of 100 µM dose of gossypin had similar effects to the routine cisplatin administration, caused a significant decrease in cell proliferation, and increased apoptosis in the evaluations of Hoechst staining and morphology. It has been put forth that gossypin decreases the expression of CASP-3 and CASP-9 mRNA and increases the expression of NF-kB.Conclusion: Our results demonstrate that for the breast cancer cells, the 100 µM of gossypin positively affects cell death pathways due to apoptosis.

Three metal complexes with a pyridyl Schiff base: cytotoxicity, migration and mechanism of apoptosis.

Three metal complexes [CuL(NO3)]n (1), [Cd(HL)(NO3)2]n (2) and [EuL(HCOOH)(H2O)(NO3)2] (3) were synthesized with a pyridyl Schiff ligand L (N'-[(1E)-pyridin-2-ylmethylidene]pyridine-4-carbohydrazide). A crystallographic study revealed that complexes 1 and 2 have a chain structure, and complex 3 is a zero-dimensional monomer. In vitro cytotoxicity studies showed that complex 2 had the best antiproliferative activity against SMMC-7721 cells and complex 3 had the best antiproliferative activity against MDA-MB-231 cells with single-digit IC50 values, both exceeding those of the control drug cisplatin by far. The cell invasion and migration ability through the transwell assay and wound-healing assay showed that the selected complexes could inhibit the invasion and migration of cancer cells. The Hoechst staining assay and ROS generation assay with SMMC-7721 cells indicated that the cytotoxic effects of complex 2 involved apoptosis induction through ROS accumulation. The apoptosis-inducing and cell cycle arrest effects of complex 2 on SMMC-7721 cells indicated that the antitumor effect was achieved through apoptosis induction and inhibition of DNA synthesis by blocking the G0/G1 phase of the cell cycle. In addition, complex 2 showed significant inhibition against B. dysentery with an inhibition circle diameter of 24 mm.

Tert-Butylhydroquinone alleviates insulin resistance and liver steatosis in diabetes.

OBJECTIVES:This work aimed to determine tert-Butylhydroquinone (TBHQ)'s effects on insulin resistance (IR) and liver steatosis in diabetic animals and to explore the underpinning mechanisms.MATERIALS AND METHODS:Male ApoE-/-mice underwent streptozocin (STZ) administration while receiving a sucrose/fat-rich diet for type 2 diabetes mellitus (T2DM) establishment. This was followed by a 6-week TBHQ administration. Body weight, fasting (FBG) and postprandial (PBG) blood glucose amounts, and insulin concentrations were measured, and the oral glucose tolerance test (OGTT) was carried out. Hematoxylin and eosin (H and E) staining and immunoblot were carried out for assessing histology and protein amounts in the liver tissue samples. In addition, cultured HepG2 cells were administered HClO and insulin for IR induction, and immunoblot was carried out for protein evaluation. Finally, the cells were stained with the Hoechst dye for apoptosis evaluation.RESULTS:The model animals showed T2DM signs, and TBHQ decreased FBG, ameliorated glucose tolerance and reduced liver steatosis in these animals. In addition, TBHQ markedly upregulated AMPKα2, GLUT4 and GSK3 β, as well as phosphorylated PI3K and AKT in the liver of mice with T2DM. In agreement, TBHQ decreased HClO-and insulin-related IR in cells and suppressed apoptosis through AMPKα2/PI3K/AKT signaling.CONCLUSIONS:TBHQ alleviates IR and liver steatosis in a mouse model of T2DM likely through AMPKα2/PI3K/AKT signaling.