Occurrence Of Apoptosis Research Articles

Melatonin protects against maternal diabetes-associated meiotic defects by maintaining mitochondrial function

Maternal diabetes has been widely reported to adversely affect oocyte quality. Although various molecules and pathways may be involved in this process, strategies to prevent maternal diabetes-induced deterioration of oocyte quality remain unexplored. Melatonin is synthesized by the pineal gland and has been shown to have beneficial effects on oocyte quality owing to its antioxidative function. In the present study, we found that the exposure of oocytes of diabetic mice to melatonin, in vitro, alleviated aberrant oocyte maturation competence. Notably, melatonin supplementation attenuated defects in spindle organization and chromosome alignment by mediating the expression of TPX2 and pericentrin localization. Importantly, melatonin eliminated the accumulation of reactive oxygen species and increased the cytosolic Ca2+ levels in diabetic oocytes by maintaining mitochondrial function. Moreover, the occurrence of autophagy and apoptosis was reversed in diabetic oocytes after melatonin exposure via decreased LC3β expression. Collectively, our findings provide evidence that melatonin supplementation can protect oocytes from maternal diabetes-related meiotic defects and poor egg quality, providing a potential strategy for improving oocyte quality in assisted reproductive technologies.

PANoptosis-like cell death in ischemia/reperfusion injury of retinal neurons.

PANoptosis is a newly identified type of regulated cell death that consists of pyroptosis, apoptosis, and necroptosis, which simultaneously occur during the pathophysiological process of infectious and inflammatory diseases. Although our previous literature mining study suggested that PANoptosis might occur in neuronal ischemia/reperfusion injury, little experimental research has been reported on the existence of PANoptosis. In this study, we used in vivo and in vitro retinal neuronal models of ischemia/reperfusion injury to investigate whether PANoptosis-like cell death (simultaneous occurrence of pyroptosis, apoptosis, and necroptosis) exists in retinal neuronal ischemia/reperfusion injury. Our results showed that ischemia/reperfusion injury induced changes in morphological features and protein levels that indicate PANoptosis-like cell death in retinal neurons both in vitro and in vivo. Ischemia/reperfusion injury also significantly upregulated caspase-1, caspase-8, and NLRP3 expression, which are important components of the PANoptosome. These results indicate the existence of PANoptosis-like cell death in ischemia/reperfusion injury of retinal neurons and provide preliminary experimental evidence for future study of this new type of regulated cell death.

Protective effects of estrogen modified hBMSC on HG-induced injury of vascular endothelial cells

Objective: To investigate the protective effects and potential mechanisms of estrogen modified human bone marrow mesenchymal stem cells (hBMSC) on high glucose (HG)-induced injury of vascular endothelial cells. Methods: hBMSCs were cultured under 30 mmol/l glucose to establish a high glucose model (HG), and then were divided into four groups as following: HG group (HG control, without any treatment), HG+E2 group (cells were treated with 20 μmol/L estrogen), HG+E2+ Triciribine group (cells were pretreated with 5 μmol/L protein kinase B (PKB/Akt) inhibitor for 45 min, and then modified by 20 μmol/L estrogen), and NG group (cells were cultured under normal conditions). After 12 h treatment, the cell viability of hBMSC was detected by CCK8 assay, and the contents of NO, VEGF and IL8 in the supernatant of cultured medium in each group were detected by nitrate reductase and ELISA assay (n=6). After 48 h, the expression levels of endothelial nitric oxide synthase (eNOS) and phosphorylated eNOS (p-eNOS) were detected by Western blot (n=3). In addition, the cell supernatant of each group was further extracted as conditioned medium to culture HUVECs, and the cells were subsequently divided into HG-CM group (HUVECs were treated with HG group's conditioned medium), HG+E2-CM group (HUVECs were treated with HG+E2 group's conditioned medium), HG+E2+Triciribine-CM group (HUVECs were treated with HG+E2+ Triciribine group's conditioned medium) and HG-H group (HUVEC were cultured under HG condition, which were treated with final concentration 30 mmol/l glucose). The cell viability of HUVECs in each group was detected by CCK8 assay after 12 h cultured (n=6). After 24 h treatment, the apoptosis rate of HUVECs in each group was detected by flow cytometry (n=3). Furthermore, the migration rate of HUVECs in each group was observed by wound healing assay after 48 h cultured (n=3). Results: Compared with NG group, the cell viability and eNOS protein phosphorylation level of hBMSC in HG group and the contents of NO, VEGF and IL-8 in the supernatant of cultured medium were decreased (P<0.05). Compared with HG group, the cell viability and eNOS protein phosphorylation level in HG+E2 group and the contents of NO, VEGF and IL-8 in cultured medium supernatant were increased significantly (P<0.05), whereas pre-treatment of hBMSC cells with a Akt inhibitor Triciribine, the above indexes showed reverse changes (P<0.05). Furthermore, compared with HG-CM group, the cell viability and migration ability (P<0.05) of HUVECs in HG+E2-CM group were increased significantly (P<0.05), and the proportion of apoptosis was decreased (P<0.05). While compared with HG+E2-CM group, the cell viability and migration ability of HUVECs in HG+E2+Triciribine-CM group were decreased (P<0.05), and the proportion of apoptosis was increased (P<0.05). Conclusion: Estrogen may promote the secretion of NO, VEGF and IL-8 by activating the Akt/eNOS signaling pathway of hBMSC cells, increase the cell viability and migration ability of HUVECs and inhibit the occurrence of apoptosis, play a protective role against the injury of HUVECs induced by HG condition.

Decorin inhibits proliferation and metastasis in human bladder cancer cells by upregulating P21.

Migration of bladder cancer (BC) cells poses a substantial threat to human health. It is critical to elucidate the mechanism of BC invasion and progression for surgical treatment and the prognosis of patients. Decorin is of interest as an anticancer treatment that can play a vital role in regulating tumorigenesis.The effect of decorin expression on survival in clinical patients was screened and analyzed using bladder urothelial carcinoma data from the Cancer Genome Atlas (TCGA) database. The differential expression of transforming growth factor-β1 (TGF-β1) in tumors was compared against that of normal samples to analyze the correlation between them. MTT, flow cytometry, and Wound/Transwell assays were used to detect cell proliferation, cycle arrest, apoptosis, migration, and invasion.Analysis of TCGA data showed that decorin expression was significantly lower in bladder urothelial carcinoma samples than in normal tissues, while TGF-β1 expression did not change significantly. We found that decorin was correlated with TGF-β1 expression in bladder urothelial cancer. In addition, decorin blocked the G1/S phase by upregulating p21 protein and inhibiting the expression of TGF-β1 and MMP2, promoting the occurrence of apoptosis and inhibiting the proliferation of human BC T24 cells. Moreover, decorin increased the adhesion of tumor cells in vitro, and effectively inhibited cell metastasis.Decorin regulated the expression of TGF-β1 and MMP2 through p21 protein, promoted apoptosis and adhesion, and inhibited the proliferation and metastasis of BC cells.

Poly(lactic-co-glycolic acid)(PLGA)-based nanoparticles modified with chitosan-folic acid to delivery of Artemisia vulgaris L. essential oil to HT-29 cancer cells

In the present study, preparation of poly (lactic-co-glycolic acid)(PLGA) nanoparticles coated with chitosan (CS) conjugated to folic acid (FA) was applied to load Artemisia vulgaris L. essential oil (AVEO) and evaluate its anti-cancer effects. To do so, AVEO was encapsulated in PLGA-NPs by W/O method and the surface of nanoparticles was coated with CS attached to FA (AVEO-PCF-NPs). Then, the physicochemical properties of AVEO-PCF-NPs were evaluated by dynamic light scattering (DLS), Zeta potential, Fourier-transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) methods, and subsequently, HPLC technique was used to calculate the amount of AVEO encapsulation and FA binding. MTT assay, florescent staining and cell cycle analysis were used to evaluate the occurrence of apoptosis. The chorioallantoic membrane (CAM) assay was done to evaluate the effect of AVEO-PCF-NPs on angiogenesis. AVEO-PCF-NPs suppressed cancer cells with IC50 ~78 μg/ml without inhibitory effect on normal cells. Arrest of cells in the SubG1 phase and AO/PI staining results showed the pro-apoptotic effects of AVEO-PCF-NPs. Significant reduction of blood vessel in samples treated with AVEO-PCF-NPs confirmed inhibitory effects of AVEO-PCF-NPs on angiogenesis. The effect of selective toxicity and the ability to induce apoptosis and inhibit angiogenesis introduces AVEO-PCF-NPs as a suitable candidate for cancer research.

Notoginsenoside R1 alleviates spinal cord injury by inhibiting oxidative stress, neuronal apoptosis, and inflammation via activating the nuclear factor erythroid 2 related factor 2/heme oxygenase-1 signaling pathway.

The secondary injury plays a vital role in the development of spinal cord injury (SCI), which is characterized by the occurrence of oxidative stress, neuronal apoptosis, and inflammatory response. Notoginsenoside R1 (NGR1) has been involved in the modulation of antioxidative stress and anti-inflammatory response. However, its roles in SCI-induced injury are still unknown. We explored the therapeutic effect of NGR1 and its underlying mechanism after SCI by using behavioral, biochemical, and immunohistochemical techniques. The administration of NGR1 after SCI enhanced the neurological function, and mitigated tissue damage and motor neuron loss than those in SCI + vehicle group. Meanwhile, significantly increased expression of Nrf2 protein and HO-1 protein was found in the SCI + NGR1 group compared with those in the SCI + vehicle group. In addition, the inhibitory effects of oxidative stress, apoptotic neuron ratio, and neuronal inflammation in the SCI + NGR1 group can be partially reversed when the Nrf2/HO-1 signaling pathway was inhibited by ML385. Our results indicate that the administration of NGR1 can attenuate oxidative stress, neuronal apoptosis, and inflammation by activating the Nrf2/HO-1 signaling pathway after SCI, thereby improving neurological function.

Satureja khuzistanica Essential Oil-Loaded Solid Lipid Nanoparticles Modified With Chitosan-Folate: Evaluation of Encapsulation Efficiency, Cytotoxic and Pro-apoptotic Properties.

The study aimed to synthesize Satureja khuzistanica essential oil-loaded SLN nanoparticles and to modify the surface of nanoparticles with folate-bound chitosan (SEO-SCF-NPs), and finally to investigate the effects of its toxicity and pro-apoptosis. For this purpose, the SEO-SLN nanoparticles were prepared using stearic acid, lecithin, tween 80, and water by high-pressure homogenization method. After characterization by FTIR, SEM, DLS, and ZETA potential methods, its toxicity effect against normal (HFF) and cancer (MCF-7) cells were evaluated by MTT assay. The occurrence of apoptosis in MCF-7 cells was assessed by flow cytometry and molecular analysis. The obtained results revealed the formation of round nanoparticles with a size of 279.40 nm, single dispersed (PDI: 0.3) and stable (ζ–potential: +31.69 mV). SEO-SCF-NPs indicated the effect of selective toxicity against MCF-7 cells (IC50: 88 μg/ml). Molecular analysis showed that SEO-SCF-NPs could inhibit cancer cells by activating the internal pathway of apoptosis as well as cell cycle disruption. Our finding suggests that SEO-SCF-NPs is a suitable candidate for preclinical cancer studies.

Nicotinamide mononucleotide supplementation improves the quality of porcine oocytes under heat stress

BackgroundElevated ambient temperature-caused heat stress is a major concern for livestock production due to its negative impact on animal feed intake, growth, reproduction, and health. Particularly, the germ cells are extremely sensitive to the heat stress. However, the effective approach and strategy regarding how to protect mammalian oocytes from heat stress-induced defects have not been determined.MethodsGerminal vesicle (GV) porcine oocytes were cultured at 41.5 °C for 24 h to induce heat stress, and then cultured at 38.5 °C to the specific developmental stage for subsequent analysis. Nicotinamide mononucleotide (NMN) was dissolved in water to 1 mol/L for a stock solution and further diluted with the maturation medium to the final concentrations of 10 μmol/L, 20 μmol/L, 50 μmol/L or 100 μmol/L, respectively, during heat stress. Immunostaining and fluorescence intensity quantification were applied to assess the effects of heat stress and NMN supplementation on the key processes during the oocyte meiotic maturation.ResultsHere, we report that NMN supplementation improves the quality of porcine oocytes under heat stress. Specifically, we found that heat stress resulted in oocyte maturation failure by disturbing the dynamics of meiotic organelles, including the cytoskeleton assembly, cortical granule distribution and mitochondrial function. In addition, heat stress induced the production of excessive reactive oxygen species (ROS) and DNA damage, leading to the occurrence of apoptosis in oocytes and subsequent embryonic development arrest. More importantly, we validated that supplementation of NMN during heat stress restored the meiotic defects during porcine oocyte maturation.ConclusionsTaken together, our study documents that NMN supplementation is an effective approach to improve the quality of oocytes under heat stress by promoting both nuclear and cytoplasmic maturation.

Eucommia ulmoides Olive Male Flower Extracts Ameliorate Alzheimer's Disease-Like Pathology in Zebrafish via Regulating Autophagy, Acetylcholinesterase, and the Dopamine Transporter.

Alzheimer’s disease (AD) is the most prevalent neural disorder. However, the therapeutic agents for AD are limited. Eucommia ulmoides Olive (EUO) is widely used as a traditional Chinese herb to treat various neurodegenerative disorders. Therefore, we investigated whether the extracts of EUO male flower (EUMF) have therapeutic effects against AD. We focused on the flavonoids of EUMF and identified the composition using a targeted HPLC-MS analysis. As a result, 125 flavonoids and flavanols, 32 flavanones, 22 isoflavonoids, 11 chalcones and dihydrochalcones, and 17 anthocyanins were identified. Then, the anti-AD effects of the EUMF were tested by using zebrafish AD model. The behavioral changes were detected by automated video-tracking system. Aβ deposition was assayed by thioflavin S staining. Ache activity and cell apoptosis in zebrafish were tested by, Acetylcholine Assay Kit and TUNEL assay, respectively. The results showed that EUMF significantly rescued the dyskinesia of zebrafish and inhibited Aβ deposition, Ache activity, and occurrence of cell apoptosis in the head of zebrafish induced by AlCl3. We also investigated the mechanism underlying anti-AD effects of EUMF by RT-qPCR and found that EUMF ameliorated AD-like symptoms possibly through inhibiting excessive autophagy and the abnormal expressions of ache and slc6a3 genes. In summary, our findings suggested EUMF can be a therapeutic candidate for AD treatment.

Osthole Induces Apoptosis and Caspase-3/GSDME-Dependent Pyroptosis via NQO1-Mediated ROS Generation in HeLa Cells.

Osthole is a natural coumarin which has been proved to inhibit growth of cancer cells by inducing cell death, while its mechanism was considered to be just caused by apoptosis. In our study, we found that osthole activated not just apoptosis, but also pyroptosis which is a form of regulated cell death accompanied by loss of cell membrane integrity and lactate dehydrogenase (LDH) release. Caspase-3 is a key protein of apoptosis as well as pyroptosis. The apoptosis and pyroptosis induced by osthole were all inhibited by irreversible caspase-3 inhibitor Z-DEVD-FMK. Meanwhile, knockdown of gasdermin E (GSDME) only reduced the osthole-induced pyroptosis but did not affect the occurrence of apoptosis. Our proteomic analysis revealed that the expression of NAD(P)H: quinone oxidoreductase 1 (NQO1) was decreased in osthole-treated cells. Moreover, NQO1 inhibition by osthole induced the overproduction of reactive oxygen species (ROS), as well as apoptosis and pyroptosis. ROS inhibitor N-Acetyl-L-cysteine (NAC) not only reduced osthole-induced apoptosis but also reversed its effect on the pyroptosis. Meanwhile, knockdown of NQO1 by si-NQO1 or its inhibitor dicoumarol (DIC) not only enhanced ROS generation but also strengthened the GSDME-mediated pyroptosis. Finally, we demonstrated that osthole inhibited tumor growth and the expression of NQO1 in a HeLa xenograft mode. Similar to the results in vitro, osthole stimulated the activation of caspase-3, PARP, and GSDME in vivo. Taken together, all these data suggested that osthole induced apoptosis and caspase-3/GSDME-mediated pyroptosis via NQO1-mediated ROS accumulation.

Benzene exposure causes structural and functional damage in rat ovaries: occurrence of apoptosis and autophagy.

Studies to date have provided evidence for damage that can occur from hydrocarbon benzene on different tissues/organs. However, little is known regarding the possible influence of this hydrocarbon on female reproduction. In this study, female Wistar rats were treated with low (2000ppm), middle (4000ppm), and high (8000ppm) doses of benzene by inhalation for 30min daily for 28days. Benzene exposure adversely affected ovarian function and structure by inducing histopathological changes and altering reproductive steroid hormone release. In addition, benzene-exposed ovaries exhibited increased TMR red fluorescent signals at middle and high doses, revealing significant apoptosis. Interestingly, the investigation of the autophagic protein marker LC3 showed that this protein significantly increased in all benzene-treated ovaries, indicating the occurrence of autophagy. Moreover, ovaries from benzene-treated groups exhibited differential regulation of several specific genes involved in ovarian folliculogenesis and steroidogenesis, including the INSL3, CCND1, IGF-1, CYP17a, LHR, ATG5, and GDF9 genes.

Role of VEGFB in electrical pulse stimulation inhibits apoptosis in C2C12 myotubes

Skeletal muscle is the major effector organ for exercise. It has been proposed that VEGFB is significantly related to apoptosis in various cell types but not yet in skeletal muscle. We hypothesize that the decrease of VEGFB in skeletal muscle participates in the occurrence of skeletal muscle apoptosis and that exercise inhibits apoptosis by elevating the expression of VEGFB in skeletal muscle cells. Based on this hypothesis, we developed in vitro experiments to mimic the effect of exercise through electrical pulse stimulation (EPS) to observe the effect of EPS on apoptosis and the change in VEGFB expression in differentiated myotubes. In addition, we employed RNA interference to explore whether VEGFB is directly involved in the regulation of myotube apoptosis during EPS. Our results showed that exogenous VEGFB167 significantly inhibited C2C12 myotube apoptosis induced by TNF-α treatment and that endogenous VEGFB in differentiated C2C12 myotubes was significantly upregulated by EPS. In addition, EPS significantly changed the expression of the apoptotic indicators Bax and Bcl-2 at the mRNA level and downregulated the protein expression of cleaved caspase-3. The antiapoptotic effect of EPS weakened substantially as VEGFB in C2C12 myotubes was inhibited. Taken together, these results indicate that exercise-like EPS inhibits apoptosis by increasing the expression of C2C12 myotube-derived VEGFB.

Inorganic mercury induces liver oxidative stress injury in quails by inhibiting Akt/Nrf2 signal pathway

Mercury is widely used in human life and production. Excessive mercury can accumulate in livestock and poultry through air, drinking water, and food, which causes a serious threat to natural environment and human food safety. However, the exact mechanism of hepatotoxicity caused by mercuric chloride (HgCl2) is still unclear. The purpose of this study is to explore the mechanism of HgCl2 poisoning in the liver. Based on the above background, 28 quails were randomly and equally divided into 4 groups to establish a hepatotoxicity model by oral administration of HgCl2 once daily (0 mg/kg, 2 mg/kg, 4 mg/kg, and 8 mg/kg, respectively). The experiment period was 84 d. The assay results of blood routine and biochemical indexes showed liver injury in quails. Further research found that mRNA and protein levels of nuclear factor-erythroid related factor 2 (Nrf2) and its key promoter protein kinase B (Akt) were significantly decreased, suggesting HgCl2 treatment led to liver oxidative stress injury in a dose-dependent manner by inhibiting the antioxidant signal pathway Akt/Nrf2. Besides, the increase of nuclear factor-κB and tumor necrosis factor-alpha levels as well as the activation of tumor protein 53 indicated the occurrence of inflammation and apoptosis. Moreover, the inhibition of multidrug resistance protein 2 (Mrp2) which participates in HgCl2-mediated toxicant metabolism resulted in toxicity accumulation. Thus, a vicious circle was formed to further aggravate the liver injury. In conclusion, our results demonstrated that HgCl2 leads to liver inflammation, apoptosis, and detoxification dysfunction by inhibiting the antioxidant pathway Akt/Nrf2. Among them, Akt and Mrp2 may be effective therapeutic targets for the treatment of HgCl2-triggered hepatotoxicity.

Zoledronic Acid use and Adverse Drug Reaction (ADR)

Bone is the part of body that most common for metastasis solid tumors. Bisphosphonates are an effective treatment for preventing bone damage events and improve quality of life patients. Zoledronic acid (ZOL) is the third generation of nitrogen-containing bisphosphonates (NBPs) that can inhibit osteoclasts in the process of bone resorption. Mechanism action of this drug related to affinity with hydroxyapatite crystals of bone, through a high bone turnover result in reducing speed of bone remodeling. This drug has higher affinity compared with other bisphosphonates for bonding hydroxyapatite . Zoledronic acid internalizes into osteoclasts, which disrupts the metabolic process and function of osteoclast cells and accelerates the occurrence of apoptosis. In the other hand it has adverse drug reactions mild to moderate which have reported but the mechanism still unclear such as hypocalcaemia and secondary hypophosphatemia, acute phase response (APR), renal toxicity, and osteonecrosis of the jaw (ONJ). Adverse drug reaction can cause harm to patient indeed arise death, highly recommended to offer monitoring for patients.

Effects of nitric oxide synthase inhibitor on mitochondria apoptosis and meat quality in postmortem Gannan yak (Bos grunniens) meat.

This research aimed to explore the effects of the nitric oxide synthase (NOS) inhibitor (L-NAME) on mitochondria apoptosis in postmortem Gannan yak (Bos grunniens) longissimus dorsi (LD) muscle and to explore its effect on meat quality further. The Gannan yak meat samples were treated with the control group (0.9% NaCl) and L-NAME (20, 60, and 100 mM) for 24 h and then stored for 0, 1, 3, 5, and 7 days at 4°C. NOS activity and NO content were investigated, and the parameters of mitochondrial apoptosis of the postmortem Gannan yak meat were determined. Meanwhile, the meat quality such as the centrifugation loss, meat color, and myofibril fragmentation index (MFI) was evaluated. The results indicated that after treatment with L-NAME, NOS activity and NO content decreased, causing mitochondrial membrane damage, Bax protein, and Cyt-c levels increased, and resulted in increased activities of caspase-9 and -3, promoting the occurrence of mitochondrial apoptosis. Furthermore, it increased the tenderness and water retention of Gannan yak meat. The results indicated that NOS inhibitor played a regulatory role in postmortem Gannan yak meat quality by regulating mitochondria apoptosis during postmortem aging. PRACTICAL APPLICATIONS: The meat's tenderness is often considered the most important factor affecting consumers' willingness to repurchase. The relationship of caspases and MFI suggested that L-NAME played a regulatory role in postmortem Gannan yak meat quality by regulating mitochondria apoptosis during postmortem aging. This study provides valuable information for the development of the Gannan yak economy in Tibetan areas.

Temporal changes in cortical microporosity during estrogen deficiency associated with perilacunar resorption and osteocyte apoptosis: A pilot study

Osteocytes can actively regulate bone microporosity, through either perilacunar resorption or micropetrosis following apoptosis. Osteocyte apoptosis is more prevalent in estrogen deficiency and changes in the lacunar-canalicular network of osteocytes have been reported. Temporal changes in bone mineralisation and osteocytes cellular strains occur, which might be associated with osteocyte-driven microporosity changes, although time dependant changes in bone microporosity are not yet fully understood. In this pilot study we conducted micro-CT analysis, backscatter electron imaging and histological analysis of femoral cortical bone form an ovariectomized rat model of osteoporosis to investigate whether estrogen deficiency causes temporal changes in lacunar and vascular porosity. We also assessed MMP14 expression, lacunar occupancy and mineral infilling, as indicators of perilacunar resorption and micropetrosis. We report temporal changes in cortical microporosity in estrogen deficiency. Specifically, canalicular and vascular porosity initially increased (4 weeks post-OVX), coinciding with the period of rapid bone loss, whereas in the longer term (14 weeks post-OVX) lacunar and canalicular diameter decreased. Interestingly, these changes coincided with an increased prevalence of empty lacunae and osteocyte lacunae were observed to be more circular with a mineralised border around the lacunar space. In addition we report an increase in MMP14+ osteocytes, which also suggests active matrix degradation by these cells. Together these results provide an insight into the temporal changes in cortical microporosity during estrogen deficiency and suggest the likelihood of occurrence of both perilacunar resorption and osteocyte apoptosis leading to micropetrosis. We propose that microporosity changes arise due to processes driven by distinct populations of osteocytes, which are either actively resorbing their matrix or have undergone apoptosis and are infilling lacunae by micropetrosis.

Comprehensive Analysis of the Potential Immune-Related Biomarker ATG101 that Regulates Apoptosis of Cholangiocarcinoma Cells After Photodynamic Therapy.

Autophagy related gene 101 (ATG101) plays a significant role in the occurrence and development of tumours by responding to stress. Our research aims to illustrate the correlation between the expression of ATG101 and tumor prognosis and its potential role and mechanism in tumor immunity and photodynamic therapy (PDT). First, integrated analysis of The Cancer Genome Atlas and Genotype-Tissue Expression portals were used to analyse the expression of ATG101. Then, Kaplan–Meier curves was applied in cholangiocarcinoma (CHOL) and liver hepatocellular carcinoma (LIHC) datasets for survival analysis. Next, the relationship between ATG101 expression and six immune cells, the immune microenvironment and immune checkpoints was analysed. Besides, the relationship between the expression of ATG101 and methyltransferase. GSEA was used to study the function and the related transcript factors of ATG101 in CHOL and LIHC. The effect of PDT on ATG101 was verified by microarray, qPCR and western blot. Then the effect of ATG101 and its regulatory factors on apoptosis were verified by siRNA, lentivirus transfection and Chip-qPCR. Comprehensive analysis showed that ATG101 was overexpressed in different tumours. Kaplan–Meier curves found that ATG101 was associated with poor prognosis in tumours (including CHOL and LIHC). We found that ATG101 can be used as a target and prognostic marker of tumour immunotherapy for different tumours. We also found that ATG101 regulates DNA methylation. GSEA analysis showed that ATG101 may play a critical role in CHOL and LIHC. Subsequent validation tests confirmed that the up-regulated ATG101 after PDT treatment is not conducive to the occurrence of apoptosis of cholangiocarcinoma cells. The high expression of ATG101 may be induced by the early stress gene EGR2. Our study highlights the significance of ATG101 in the study of tumour immunity and photodynamic therapy from a pan-cancer perspective.

Programmed cell death might involve in progress of wounding induced agarwood formation in stems of Aquilaria sinensis.

Agarwood, a non-timber fragrant wood, is derived from wounded Aquilaria trees (Thymelaeaceae) and is widely used in traditional medicine, incense and perfume. Agarwood-like substances and programmed cell death (PCD) can be induced by wounding signals in the suspension cells and aerial roots of Aquilaria sinensis. In this study, the dynamic process of wound-induced agarwood formation in stems of A. sinensis was observed, and the occurrence of PCD was synchronously detected using techniques such as 4',6-diamidino-2-phenylindole and dUTP nick-end labeling staining. The results showed that the wounding was a induce signal for agarwood resin formation, meanwhile might induce PCD. Interxylary phloem and xylem ray were the main sites of agarwood resin formation and PCD occurrence. There might be a relationship between the spatiotemporal pattern of PCD and agarwood resin formation: more severe PCD corresponded to a higher rate of resin formation but a shorter resin formation time; conversely, slower PCD progression corresponded to a lower rate of resin formation but a longer resin formation time. Our findings are the first to demonstrate that PCD might occur in the process of wound-induced agarwood formation at the tree level, and the spatiotemporal pattern is closely related to the formation of agarwood resin. This study provides valuable insight for further studies on the relationship between PCD and agarwood formation. HIGHLIGHTS: Programmed cell death (PCD) might occur in the process of wound-induced agarwood formation at the tree level. Interxylary phloem and xylem ray were the main sites of agarwood resin formation and PCD occurrence. Spatiotemporal pattern of PCD might have a strong impact on agarwood resin formation.

Doxorubicin Induces Necroptosis in Young Mice

Despite its potency as a chemotherapeutic drug, the prevalence of cardiotoxicity resulting from doxorubicin treatment in childhood survivors of cancer is a major concern. While the occurrence of apoptosis has been well ascertained in doxorubicin induced cardiotoxicity (DIC) by several studies; however, the presence of necroptosis in young DIC remains unknown. We hypothesize that doxorubicin will induce necroptotic cell death and adverse cardiac remodeling in young mice. Therefore, in this study we investigated the possible occurrence of necroptotic cell death in DIC in young mice in‐vivo to delineate underlying pathological mechanism.C57BL6 mice (6 + 2 weeks of age) were divided into two groups: Control (mice were given 0.9% saline) and Dox (drug was administered at 2.5mg/kg body weight (BW) for six i.p injection, with a cumulative dose of 15mg/kg BW). At day 14, heart function was examined using echocardiography, mice were euthanized. Heart tissues were collected and analyzed for necroptosis, adverse cardiac remodeling using immunohistochemistry, and histology. Upon analysis, we observed that Dox treatment significantly (p&lt;0.05) increased the expression of necroptotic markers [Receptor interacting Serine/Threonine kinase 1 and 3 (RIPK1, RIPK3) and Mixed Lineage kinase domain‐Like (MLKL)] as confirmed by immunohistochemistry. Furthermore, histological analysis showed adverse cardiac remodeling with significant increase (p&lt;0.05) in cardiac hypertrophy and fibrosis in Dox group compared to control. Moreover, a significant decrease (p&lt;0.05) in heart function was observed in Dox treated animals compared to control. In conclusion the results of our study suggest that doxorubicin, induces necroptotic cell death which further leads to adverse cardiac remodeling in Dox treated young mice.

Cold Atmospheric Plasma-Activated Media Improve Paclitaxel Efficacy on Breast Cancer Cells in a Combined Treatment Model.

Simple SummaryBreast cancer is the most commonly diagnosed cancer and the leading cause of cancer mortality in females, with chemotherapy currently the only adjunctive therapy. However, the disadvantages of chemotherapy are represented by toxicity/side effects, and for many women the benefit is uncertain. Cold atmospheric plasma in in vitro and in vivo experiments proved that it is able to selectively inhibit the growth of cancer cells in various cancer types, with less deleterious side effects. Current guidelines include paclitaxel as a chemotherapy of choice for various types of breast cancers. The aim of the study was to evaluate in vitro, on human breast cancer cell lines, a plasma-activated media as a cytotoxic agent and as an associative agent in a combined treatment for breast cancer with paclitaxel. The collected data indicated that the plasma-activated media amplified the cytotoxic effect of paclitaxel.The use of plasma-activated media (PAM), an alternative to direct delivery of cold atmospheric plasma to cancer cells, has recently gained interest in the plasma medicine field. Paclitaxel (PTX) is used as a chemotherapy of choice for various types of breast cancers, which is the leading cause of mortality in females due to cancer. In this study, we evaluated an alternative way to improve anti-cancerous efficiency of PTX by association with PAM, the ultimate achievement being a better outcome in killing tumoral cells at smaller doses of PTX. MCF-7 and MDA-MB-231 cell lines were used, and the outcome was measured by cell viability (MTT assay), the survival rate (clonogenic assay), apoptosis occurrence, and genotoxicity (COMET assay). Treatment consisted of the use of PAM in combination with under IC50 doses of PTX in short- and long-term models. The experimental data showed that PAM had the capacity to improve PTX’s cytotoxicity, as viability of the breast cancer cells dropped, an effect maintained in long-term experiments. A higher frequency of apoptotic, dead cells, and DNA fragmentation was registered in cells treated with the combined treatment as compared with those treated only with PT. Overall, PAM had the capacity to amplify the anti-cancerous effect of PTX.