FIGURE 3 - uploaded by Caroline Bonner
Content may be subject to copyright.
AMPK mediates DN-HNF1A-induced apoptosis. A, Western blot of phospho-Thr-172 AMPK levels after treatment with doxycycline in the presence and absence of the AMPK inhibitor compound C (20 M) for 48 h. B, quantification of phospho-Thr-172 AMPK levels after treatment with doxycycline for indicated times. Western blot images were analyzed as described under " Experimental Procedures. " Data shown represent means S.E. from three independent experiments. *, p 0.05 compared with untreated controls . #, p 0.05 compared with samples treated with doxycycline alone. C, Western blot of cleaved caspase 3 levels after treatment with doxycycline in the presence and absence of compound C (20 M) for the indicated times. D, quantification of cleaved caspase 3 levels after treatment with doxycycline for the indicated times. Western blot images were analyzed as described under " Experimental Procedures. " Data shown represent means S.E. from three independent experiments. E, effect of compound C on DN-HNF1A-in- duced apoptosis. Cells were treated with doxycycline and/or compound C as indicated for 48 h and then stained with Hoechst. Three images were taken from each culture, and the number of cells with apoptotic nuclei was quantified . Data shown represent means S.E. from n 4 cultures. This experiment was repeated twice with similar results. *, p 0.05 compared with samples treated with doxycycline alone. F, effect of AMPK gene silencing on DN-HNF1A-induced apoptosis. Cells were transiently transfected with a copGFP-expressing vector containing a scrambled or AMPK siRNA sequence and subsequently treated with doxycycline and as indicated for the times shown. Approximately 35% of cells were found to be copGFP-positive. Cultures were stained with Hoechst, and three images were taken from each well. Fragmented and/or condensed nuclei in copGFP-positive cells were scored as apoptotic and expressed as a percentage of total copGFP-positive cells. Data shown represent means S.E. from n 4 cultures. *, p 0.05 compared with samples transfected with control siRNA and treated with doxycycline for 48 h.
Source publication
Heterozygous loss-of-function mutations in the hepatocyte nuclear factor 1A (HNF1A) gene result in the pathogenesis of maturity-onset diabetes-of-the-young type 3, (HNF1A-MODY). This disorder is characterized by a primary defect in metabolism-secretion coupling and decreased beta cell mass, attributed to excessive beta cell apoptosis. Here, we inve...
Contexts in source publication
Context 1
... this end, we first pharmacologically inhibited AMPK activity using compound C and examined its effects on phosphorylated AMPK and cleaved caspase 3 by Western blot. Caspase 3 is cleaved to become active, which is considered to be a primary event in the initia- tion of apoptosis. Compound C was found to inhibit DN- HNF1A-induced activation of AMPK (Fig. 3, A and B), which correlated with decreased levels of DN-HNF1A-induced cleaved caspase 3 (Fig. 3, C and D). Furthermore, the level of DN-HNF1A-induced apoptosis after 48 h was dramatically attenuated in cells co-treated with compound C (Fig. 3E). To ensure that the protective effects observed using compound C were not due to possible ...
Context 2
... on phosphorylated AMPK and cleaved caspase 3 by Western blot. Caspase 3 is cleaved to become active, which is considered to be a primary event in the initia- tion of apoptosis. Compound C was found to inhibit DN- HNF1A-induced activation of AMPK (Fig. 3, A and B), which correlated with decreased levels of DN-HNF1A-induced cleaved caspase 3 (Fig. 3, C and D). Furthermore, the level of DN-HNF1A-induced apoptosis after 48 h was dramatically attenuated in cells co-treated with compound C (Fig. 3E). To ensure that the protective effects observed using compound C were not due to possible off-target effects, we utilized siRNA technology to silence AMPK expression, which we have previ- ously used ...
Context 3
... in the initia- tion of apoptosis. Compound C was found to inhibit DN- HNF1A-induced activation of AMPK (Fig. 3, A and B), which correlated with decreased levels of DN-HNF1A-induced cleaved caspase 3 (Fig. 3, C and D). Furthermore, the level of DN-HNF1A-induced apoptosis after 48 h was dramatically attenuated in cells co-treated with compound C (Fig. 3E). To ensure that the protective effects observed using compound C were not due to possible off-target effects, we utilized siRNA technology to silence AMPK expression, which we have previ- ously used to effectively target the 1/2 subunit (17). As dem- onstrated in Fig. 3F, siRNA knockdown of AMPK was also shown to be protective against ...
Context 4
... after 48 h was dramatically attenuated in cells co-treated with compound C (Fig. 3E). To ensure that the protective effects observed using compound C were not due to possible off-target effects, we utilized siRNA technology to silence AMPK expression, which we have previ- ously used to effectively target the 1/2 subunit (17). As dem- onstrated in Fig. 3F, siRNA knockdown of AMPK was also shown to be protective against DN-HNF1A-induced apoptosis (Fig. ...
Context 5
... the protective effects observed using compound C were not due to possible off-target effects, we utilized siRNA technology to silence AMPK expression, which we have previ- ously used to effectively target the 1/2 subunit (17). As dem- onstrated in Fig. 3F, siRNA knockdown of AMPK was also shown to be protective against DN-HNF1A-induced apoptosis (Fig. ...
Similar publications
The forkhead O transcription factors (FOXO) integrate a range of extracellular signals, including growth factor signaling, inflammation, oxidative stress, and nutrient availability, to substantially alter the program of gene expression and modulate cell survival, cell cycle progression, and many yet to be unraveled cell type-specific responses. Nai...
Maturity-onset diabetes of the young type 3 (MODY3) is caused by heterozygous mutation in the HNF1A gene. Liver adenomatosis has been reported in MODY3 patients. The patient reported in this paper is a Japanese girl who first developed hepatomegaly, fatty liver, and hepatic dysfunction at age 5 years. Liver biopsy demonstrated steatosis and degener...
Hyperinsulinism (HI) is the leading cause of persistent hypoglycemia in children, which if unrecognized may lead to development delays and permanent neurologic damage. Prompt recognition and appropriate treatment of HI are essential to avoid these sequelae. Major advances have been made over the past two decades in understanding the molecular basis...
Hepatocyte nuclear factor 1-α (HNF1A)/hepatocyte nuclear factor 4-α (HNF4A) maturity-onset diabetes of the young (MODY) is frequently misdiagnosed as type 1 diabetes, and patients are inappropriately treated with insulin. Blood C-peptide can aid in the diagnosis of MODY, but practical reasons limit its widespread use. Urinary C-peptide creatinine r...
Citations
... 39,40 Among these genes, BCL2L11, belonging to the BCL-2 family and situated in the mitochondrial outer membrane, serves as a crucial apoptosis regulator. 41,42 Enhancing BCL2L11-mediated apoptosis has been shown to prevent drug resistance. 43 Additionally, BBC3 ...
Background
Globally, breast cancer constitutes the predominant malignancy in women. Abnormal regulation of epigenetic factors plays a key role in the development of tumors. Anti‐apoptosis is a characteristic of tumor cells. Therefore, exploring and identifying relevant epigenetic factors that regulate the apoptosis of tumor cells is the foundation for clarifying the pathogenesis of tumors and achieving precision antitumor therapy.
Method
This study focused on exploring the epigenetic mechanism of FOXK1 in the development of estrogen receptor‐positive (ER ⁺ ) breast cancer. We used overexpressing FLAG‐FOXK1 MCF‐7 cells to perform silver staining mass spectrometry analysis and conducted Co‐IP experiments to verify the interactions. ChIP‐seq was conducted on MCF‐7 cells to examine FOXK1's binding across the genome and its transcriptional target sites. To validate the ChIP‐seq results, qChIP, western blotting, and quantitative polymerase chain reaction (qPCR) were performed. Through TUNEL assay, cell counting assay, colony formation assay, and the mouse xenograft models, the effect of FOXK1 on breast cancer progression was detected. Finally, by analyzing online databases, the correlation between FOXK1 and the survival of breast cancer patients was examined.
Results
FOXK1 interacts with the REST/CoREST transcriptional corepression complex to transcriptionally inhibit target genes representing the apoptotic pathway. Abnormally high expression of FOXK1 prevents the apoptosis of ER ⁺ breast cancer cells in vitro and promotes ER ⁺ breast tumor progression in vivo. Furthermore, the expression of FOXK1 is negatively correlated with the survival of ER ⁺ breast cancer patients.
Conclusion
FOXK1 promotes ER ⁺ breast carcinogenesis through anti‐apoptosis and acts as a potential target for ER ⁺ breast cancer treatment.
... As a member of the BCL-2 protein family, BCL2L11 is located in the outer mitochondrial membrane, and plays an important regulatory role in mediating excitatory apoptosis, induction of gene sequence translocation, and mitochondrial depolarization (40,41). It was found that miR-29b inhibited the pro-apoptotic protein BCL2L11, Bcl-2 modifier, Bcl-2 interacting protein Harakir, and Bcl-2 binding component 3 (p53-upregulated modulator of apoptosis) during neuronal development, and played an essential role in neuronal maturation and inhibition of neuronal apoptosis (42). ...
Acute ischemic stroke (AIS) is a common acute cerebrovascular disease. Circular RNAs (circRNAs) have been demonstrated to have critical functions in a wide range of physiological processes and disorders in humans. However, their precise function in ischemic stroke (IS) remains largely unknown. The present study explored the function and potential mechanisms of circ_0000018 in AIS in vivo and in vitro. The cerebral ischemia/reperfusion injury model was established in vivo and in vitro using the oxygen-glucose deprivation (OGD/R) and transient middle cerebral artery occlusion (tMCAO) methods. Subsequently, the impact of circ_0000018 on cerebral ischemia/reperfusion injury was assessed using various techniques, including TTC staining, quantitative PCR, western blotting, cell counting kit-8 assay, Annexin V-FITC Apoptosis Detection Kit, luciferase reporter gene assays, and others. The levels of circ_0000018 were markedly increased in the OGD/R-treated neuronal cells and in a mouse model of tMCAO. The blocking of microRNA (miR)-871 by circ_0000018 promoted Bcl-2-like protein 11 (BCL2L11) expression to increase neuronal cell damage. Furthermore, circ_0000018 knockdown significantly improved neuronal cell viability and attenuated OGD/R-treated neuronal cell death. Meanwhile, circ_0000018 knockdown improved brain infarct volume and neuronal apoptosis in tMCAO mice. The present study found that circ_0000018 knockdown relieved cerebral ischemia-reperfusion injury progression in vitro and in vivo. Mechanistically, circ_0000018 regulated the levels of BCL2L11 by sponging miR-871.
... Available data on the role of AMPK in the regulation of apoptosis are controversial. While some reports suggest AMPK-dependent stimulation of apoptosis [39][40][41], several studies demonstrate an anti-apoptotic role of AMPK [22,[42][43][44]. ...
Regrowth of atrophied myofibers depends on muscle satellite cells (SCs) that exist outside the plasma membrane. Muscle atrophy appears to result in reduced number of SCs due to apoptosis. Given reduced AMP-activated protein kinase (AMPK) activity during differentiation of primary myoblasts derived from atrophic muscle, we hypothesized that there may be a potential link between AMPK and susceptibility of differentiating myoblasts to apoptosis. The aim of this study was to estimate the effect of AMPK activation (via AICAR treatment) on apoptosis in differentiating myoblasts derived from atrophied rat soleus muscle. Thirty rats were randomly assigned to the following two groups: control (C, n = 10) and 7-day hindlimb suspension (HS, n = 20). Myoblasts derived from the soleus muscles of HS rats were divided into two parts: AICAR-treated cells and non-treated cells. Apoptotic processes were evaluated by using TUNEL assay, RT-PCR and WB. In differentiating myoblasts derived from the atrophied soleus, there was a significant decrease (p < 0.05) in AMPK and ACC phosphorylation in parallel with increased number of apoptotic nuclei and a significant upregulation of pro-apoptotic markers (caspase-3,-9, BAX, p53) compared to the cells derived from control muscles. AICAR treatment of atrophic muscle-derived myoblasts during differentiation prevented reductions in AMPK and ACC phosphorylation as well as maintained the number of apoptotic nuclei and the expression of pro-apoptotic markers at the control levels. Thus, the maintenance of AMPK activity can suppress enhanced apoptosis in differentiating myoblasts derived from atrophied rat soleus muscle.
... Both changes were prevented by preoperative resistance exercise. The activation of AMPK under times of energy stress can be both neuroprotective [68] and proapoptotic [69]. In animal stroke models, acute treatment with metformin, an AMPK-α agonist, has been shown to increase infarct volume; but reduce infarct volume with chronic administration [70]. ...
Background
Postoperative neurocognitive dysfunction remains a significant problem in vulnerable groups such as the elderly. While experimental data regarding its possible pathogenic mechanisms accumulate, therapeutic options for this disorder are limited. In this study, we evaluated the neuroprotective effect of a period of preconditioning resistant training on aged mice undergoing abdominal surgery. Further, we examined the underlying mechanisms from the perspective of neuroinflammatory state and synaptic plasticity in the hippocampus.
Methods
18-month-old C57BL/6N mice were trained for 5 weeks using a ladder-climbing protocol with progressively increasing weight loading. Preoperative baseline body parameters, cognitive performance and neuroinflammatory states were assessed and compared between sedentary and trained groups of 9-month-old and 18-month-old mice. To access the neuroprotective effect of resistance training on postoperative aged mice, both sedentary and trained mice were subjected to a laparotomy under 3% sevoflurane anesthesia. Cognitive performance on postoperative day 14, hippocampal neuroinflammation, mitochondrial dysfunction and synaptic plasticity were examined and compared during groups.
Results
18-month-old mice have increased body weight, higher peripheral and central inflammatory status, reduction in muscle strength and cognitive performance compared with middle-aged 9-month-old mice, which were improved by resistance exercise. In the laparotomy group, prehabilitative resistant exercise improved cognitive performance and synaptic plasticity, reduced inflammatory factors and glial cells activation after surgery. Furthermore, resistance exercise activated hippocampal PGC-1α/BDNF/Akt/GSK-3β signaling and improved mitochondrial biogenesis, as well as ameliorated mitochondrial dynamics in postoperative-aged mice.
Conclusions
Resistance exercise reduced risk factors for perioperative neurocognitive disorders such as increased body weight, elevated inflammatory markers, and pre-existing cognitive impairment. Accordantly, preoperative resistance exercise improved surgery-induced adverse effects including cognitive impairment, synaptic deficit and neuroinflammation, possibly by facilitate mitochondrial health through the PGC1-a/BDNF pathway.
... TRAF2 promotes p53-dependent apoptosis by activating the JNK signaling cascade in cancer cells (Tsuchida et al., 2020). BCL2L11 is a member of BCL-2 family and regulates function of mitochondria Kilbride et al., 2010). BCL2L11 deletion/downregulation is found in many neoplasms and contribute to acquired drug resistance (Zhang et al., 2016). ...
Purpose: The prognosis of head and neck squamous cell carcinoma (HNSCC) is poor. Necroptosis is a novel programmed form of necrotic cell death. The prognostic value of necroptosis-associated lncRNAs expression in HNSCC has not been explored.
Methods: We downloaded mRNA expression data of HNSCC patients from TCGA databases. Prognostic lncRNAs were identified by univariate Cox regression. LASSO was used to establish a model with necroptosis-related lncRNAs. Kaplan-Meier analysis and ROC were applied to verify the model. Finally, functional studies including gene set enrichment analyses, immune microenvironment analysis, and anti-tumor compound IC50 prediction were performed.
Results: We identified 1,117 necroptosis-related lncRNAs. The Cox regression showed 55 lncRNAs were associated with patient survival (p < 0.05). The risk model of 24- lncRNAs signature categorized patients into high and low risk groups. The patients in the low-risk group survived longer than the high-risk group (p < 0.001). Validation assays including ROC curve, nomogram and correction curves confirmed the prediction capability of the 24-lncRNA risk mode. Functional studies showed the two patient groups had distinct immunity conditions and IC50.
Conclusion: The 24-lncRNA model has potential to guide treatment of HNSCC. Future clinical studies are needed to verify the model.
... MF treatment was also reported to induce apoptosis and inhibit the proliferation of bile duct cancer cells through the AMPK and insulin-like growth factor 1 receptor (IGF1) pathway [57]. AMPK was also reported to induce apoptosis in response to bio-energetic stress through its ability to activate the pro-apoptotic Bcl-2 homology domain-3-only protein (BMF) in transformed rat pancreatic β cells [58]. The strong anti-growth effects of MF on pancreatic β cells is consistent with previous work which has shown that MF treatment induces hypoxia and death in INS-1 cells by blocking the expression of the HIF-1α independent anti-hypoxia factor HNF4α [59]. ...
Purpose
Metformin (MF) intake associates with reduced levels of circulating low-density lipoprotein-cholesterol (LDL-C). This has been attributed to the activation of AMPK, which differentially regulates the expression of multiple genes involved in cholesterol synthesis and trafficking. However, the exact mechanism underlying the LDL-C lowering effect of MF remains ambiguous.
Methods
MF-treated Hep-G2 and HuH7 cells were evaluated for cell viability and the expression status of key lipid metabolism-related genes along with LDL-C uptake efficiency.
Results
MF treatment resulted in decreased expression and secretion of PCSK9, increased expression of LDLR and enhanced LDL-C uptake in hepatocytes. It also resulted in increased expression of activated AMPK (p-AMPK) and decreased expression of SREBP2 and HNF-1α proteins. Transcriptomic analysis of MF-treated Hep-G2 cells confirmed these findings and showed that other key lipid metabolism-related genes including those that encode apolipoproteins (APOB, APOC2, APOC3 and APOE), MTTP and LIPC are downregulated. Lastly, MF treatment associated with reduced HMG-CoA reductase expression and activity.
Conclusions
These findings suggest that MF treatment reduces circulating LDL-C levels by suppressing PCSK9 expression and enhancing LDLR expression; hence the potential therapeutic utility of MF in hypercholesterolemia.
... miR-9a-5p was found to negatively regulate BCL2L11 expression in HPMECs. BCL2L11, a member of the BCL-2 family, is located in the outer membrane of mitochondria and it serves as an important regulator of apoptosis (28,29). Thus, miR-9a-5p may regulate HPMECs apoptosis via negatively regulating BCL2L11 expression. ...
The aim of the present study was to investigate the function of long non-coding RNA taurine-upregulated gene 1 (lncRNA TUG1) in chronic obstructive pulmonary disease and further assess the underlying molecular mechanisms. Flow cytometry analysis was performed to detect cell apoptosis of human pulmonary microvascular endothelial cells (HPMECs) treated with 1% cigarette smoke extract (CSE). The activity of caspase-3 was measured using a Caspase-3 Activity assay kit and the protein expression of cleaved caspase-3, caspase-3 and Bcl-2 like 11 (BCL2L11) were measured using western blotting. Reverse transcription-quantitative PCR (RT-qPCR) was performed to measure the expression of TUG1 mRNA levels in the treated cells. The association between TUG1, the miR-9a-5p/BCL2L11 axis and with miR-9a-5p were predicted and verified using a dual luciferase reporter assay system. The mRNA expression of miR-9a-5p and BCL2L11, and the transfection efficiency were measured by RT-qPCR. The results showed that CSE induced cell apoptosis and increased lncRNA TUG1 expression in HPMECs. CSE significantly reduced the expression of miR-9a-5p in HPMECs compared with the control group. TUG1-short hairpin RNA relieved cell apoptosis induced by CSE by upregulating miR-9a-5p in HPMECs. The present study predicted and verified that BCL2L11 is a direct target of miR-9a-5p. The mRNA expression of BCL2L11 was increased in HPMECs following CSE treatment compared with the control group. miR-9a-5p mimic and BCL2L11-plasmid markedly increased the expression of miR-9a-5p and BCL2L11, respectively. miR-9a-5p mimic reversed the increase in cell apoptosis induced by CSE by inhibiting BCL2L11 expression in HPMECs. To conclude, the present study demonstrated that lncRNA TUG1 exerted roles in cell apoptosis induced by CSE through modulating the miR-9a-5p/BCL2L11 axis.
... Moreover, there is evidence that miR-24-BCL2L11 axis mediates the survival of gastric cancer cells [11]. BCL2L11 is a pro-apoptotic factor in the B cell lymphoma-2 (Bcl-2) family regulating translocation of apoptosis-inducing factor and mitochondrial depolarization [12,13]. It has also been reported to mediate the proliferation and apoptosis of vascular smooth muscle cells in CHD [14]. ...
Coronary heart disease (CHD) is a fatal disease associated with coronary atherosclerosis. Although triptolide (TTL) has been reported to protect against CHD, the mechanism has not yet been determined. This study intended to explore its molecular regulation mechanism in CHD. It is shown in this study that TTL contributed to the proliferation and migration of in vitro cell models of CHD (endothelial cells) and the inhibition of apoptosis, and had an improvement effect on apoptosis factors and endoplasmic reticulum stress (ERS). From its mechanisms, TTL evidently downregulates miR-24-3p which is elevated in CHD, and evidently upregulates BCL2-like 11 (BCL2L11) which is suppressed in CHD, as well as affects the activation of peroxisome proliferator-activated receptors (PPARs)-Peroxisome proliferator activated receptor-γ co-activator-1α (PGC-1α) pathway of nuclear receptor transcription factors. In addition, miR-24-3p-BCL2L11-PPARs-PGC1α axis regulates protective effects of TTL against CHD.
... In addition, NADPH levels decline leads to increase of intracellular ROS level, and the cancer cells may be more sensitive to apoptosis than other cells under these conditions [170]. However, the energetic stress and reduced endogenous ATP level activates the AMPK, the sensor of ATP level [171]. In turn, AMPK reduces acetyl-CoA carboxylase which is essential for fatty acid synthesis (FAS) and for the reduction of fatty acid oxidation (FAO) [172]. ...
Reactive oxygen species (ROS) are generally recognized as major products in the cells that are produced during oxygen utilization, cellular breakdown, and the incomplete degradation of oxygen molecules. Deep research over the last two decades shows that ROS is the key factor for the initiation and regulation of cancer signaling pathways. The unique charisma of cancer cells is their ability to produce an elevated amount of oxidative stress and their requirement for an antioxidant defence system that differentiates the cancer cells from the normal cells. The low and high level of ROS controls the various signaling pathways associated with tumorigenesis, oncogenesis, angiogenesis, autophagy, lipid, glucose metabolism, etc. The elevated ROS level is a well-known hallmark for various cancer resulting in apoptosis by regulating pro-tumorigenic and anti-tumorigenic signaling pathways. In the meantime, the reduced ROS level contributes to cell survival, invasion, metastasis, oncogenesis, etc. The significant role of this double-edged sword ROS in cellular homeostasis, as well as carcinogenesis, will be discussed in this review. Moreover, the present review deals with the various intracellular and extracellular sources of ROS generation, the significant role of ROS, and provides a brief overview of the ROS signaling pathway in cancer.
... Our study showed that miR-132-3p is upregulated in endometriosis tissue samples and fortunately, reciprocal repression between LINC00261 and miR-132-3p was observed in present study. BCL2L11 (also known as BIM) is a member of BCL-2 family and is located in the outer membrane of mitochondria, where this protein acts as an important regulator that mediates apoptosis [27][28][29] and as an dual-agent that regulates autophagy in drug resistance [30]. The anti-apoptotic BCL2 members have multiple domains; while the pro-apoptotic members of BCL2 family, including BCL2L11, are BH3-domain-only proteins [31]. ...
Endometriosis is a benign disease but manifests with malignant features and limited treatment options. Women with endometriosis should not be ignored or patronized by the medical profession and society. In this regard, a major cultural change and searching for the optimum therapeutic regimen from multiple perspectives is needed in China even in the whole world. Long non-coding RNAs are crucial for various human diseases while its potential functions and mechanisms are largely unknown in endometriosis. LINC00261 was significantly downregulated in endometriosis tissues and our study indicated that it suppresses proliferation and invasion of endometriosis cells functionally in vitro. Insights of the mechanism of competitive endogenous RNAs were obtained from bioinformatic analysis, RIP, RNA pull-down and luciferase assays, which further confirmed that LINC00261 functions as a molecular sponge to regulate BCL2L11 expression by binding to miR-132-3p directly. These data defined LINC00261/miR-132-3p/BCL2L11 regulatory networks may be a novel therapeutic target for endometriosis.
