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| Mefloquine remodels proteome of KYSE150 cells. MS analysis of global protein changes in KYSE150 cells after MQ (10 µM) treatment for 24 h compared with DMSO treatment. (A) Heat map representation of differentially expressed proteins between DMSO and MQ (10 µM) groups. The color key shows the log2 transformed protein amount ratios; (B) WoLF PSORT software analysis of the sub-cellular localization of differentially changed proteins; (C) Gene ontology term enrichment classification of differentially changed proteins according to cellular component and molecular function, with the top 4 related components and functions shown; (D) KEGG pathway enrichment map of down-regulated pathways (blue bars). Data was shown as -Log10 (Fisher's exact test p-value); (E) Venn diagram indicating the overlap between the 5 downregulated signaling pathways. Four overlapping proteins were listed; (F) Averaged quantitative kinase data from the full proteome, ranked according to their Log2 FC between the DMSO and MQ-treated cells, and the locations of SDHC, SDHD, MTCO3, and NDUFV3 were shown; (G) Western blot for SDHC, SDHD, MTCO3, and NDUFV3 in KYSE150 cells after DMSO or MQ (10 µM) treatment.
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Esophageal squamous cell carcinoma (ESCC) has a worldwide impact on human health, due to its high incidence and mortality. Therefore, identifying compounds to increase patients' survival rate is urgently needed. Mefloquine (MQ) is an FDA-approved anti-malarial drug, which has been reported to inhibit cellular proliferation in several cancers. Howev...
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... treatment caused a series of proteins to be differentially expressed in KYSE150 cells (Figure 3A). WOLF PSORT software was used to classify the sub cellular localization of the differential expression proteins, showing that 26% of the differentially expressed proteins localized to the mitochondria ( Figure 3B). ...
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... treatment caused a series of proteins to be differentially expressed in KYSE150 cells (Figure 3A). WOLF PSORT software was used to classify the sub cellular localization of the differential expression proteins, showing that 26% of the differentially expressed proteins localized to the mitochondria ( Figure 3B). Although plasma membrane was more affected (28%) compared with mitochondria (26%). ...
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... plasma membrane was more affected (28%) compared with mitochondria (26%). Gene ontology analysis also showed that the mitochondria were more sensitive to MQ treatment, based on cellular component and molecular function classification ( Figure 3C). The cellular components identified as being affected by MQ treatment included the mitochondrial membrane, mitochondrial envelope, and mitochondrion, and affected molecular functions included succinate dehydrogenase activity and electron carrier activity. ...
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... cellular components identified as being affected by MQ treatment included the mitochondrial membrane, mitochondrial envelope, and mitochondrion, and affected molecular functions included succinate dehydrogenase activity and electron carrier activity. Meanwhile, a KEGG pathway enrichment map showed the top five signaling pathways (Figure 3D), which were downregulated after MQ treatment. Venn diagram analyses indicated that mitochondrial proteins succinate dehydrogenase complex subunit C (SDHC), succinate dehydrogenase complex subunit D (SDHD), mitochondrially encoded cytochrome c oxidase III (MTCO3), and NADH: ubiquinone oxidoreductase subunit V3 (NDUFV3) were all involved in these pathways ( Figure 3E). ...
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... a KEGG pathway enrichment map showed the top five signaling pathways (Figure 3D), which were downregulated after MQ treatment. Venn diagram analyses indicated that mitochondrial proteins succinate dehydrogenase complex subunit C (SDHC), succinate dehydrogenase complex subunit D (SDHD), mitochondrially encoded cytochrome c oxidase III (MTCO3), and NADH: ubiquinone oxidoreductase subunit V3 (NDUFV3) were all involved in these pathways ( Figure 3E). In addition, the extensive protein reprogramming highlights a visible decrease of SDHC, SDHD, MTCO3, and NDUFV3 in MQ-treated cells (Figure 3F). ...
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... diagram analyses indicated that mitochondrial proteins succinate dehydrogenase complex subunit C (SDHC), succinate dehydrogenase complex subunit D (SDHD), mitochondrially encoded cytochrome c oxidase III (MTCO3), and NADH: ubiquinone oxidoreductase subunit V3 (NDUFV3) were all involved in these pathways ( Figure 3E). In addition, the extensive protein reprogramming highlights a visible decrease of SDHC, SDHD, MTCO3, and NDUFV3 in MQ-treated cells (Figure 3F). A separate table contains a list of the top down differentially regulated genes affected by the treatment was also shown in Table S1. ...
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... separate table contains a list of the top down differentially regulated genes affected by the treatment was also shown in Table S1. Western blot also verified that protein levels of SDHC, SDHD, MTCO3, and NDUFV3 were decreased in KYSE150 cells after MQ treatment ( Figure 3G). Therefore, based on the results of sub cellular localization classification, Gene ontology analysis and KEGG pathway enrichment, mitochondria dysfunction may play a vital role in MQ-induced anti-tumor effect. ...
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... cases of ESCC PDXs were chosen for experiment, including case EG59, case EG60, case EG20, and case EG84. The protein levels of SDHC are high in case EG59 and EG60, while the protein levels are low in case EG20 and EG84 (Figure 6A), while not the same with SDHD, MTCO3 and NDUFV3 (Figure S3). Mice were treated with solvent (0.9% normal saline) or MQ (50 or 200 mg/kg) by oral gavage, once a day. ...
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The emergence of drug-resistant malaria parasites to artemisinin and its partner drugs highlights the need to increase the arsenal of new antimalarials with novel mechanisms of action. To help achieve this aim, this study tested the potency of three Malaria Box compounds (MMV006087, MMV085203, and MMV008956) against five laboratory strains and twen...
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... In keeping with an impact on ATP production, disruption to the MMP was MQ concentration dependent 6 h post-exposure, with significant changes detected after exposures of 3.3 and 10 µM for 24 and 48 h, respectively ( Figure 5). In support of the impact of MQ on mitochondria, recent studies have shown that 10 µM MQ reduces the cell number in KYSE150 cells (an oesophageal squamous cell carcinoma cell line), reduces the NAD + /NADH ratio, and also influences the mitochondrial proteome, including key proteins involved in the oxidative phosphorylation reactions that lead to ATP production [45]. ...
Mefloquine (MQ) is a quinoline-based anti-malarial drug used for chemoprophylaxis or as a treatment in combination with artesunate. Although MQ has clear anti-Plasmodium falciparum properties, it can induce neurotoxicity and undesired neuropsychiatric side effects in humans. Hence, this study aimed to characterize the neurotoxicity of MQ using human neuroblastoma SH-SY5Y cells. The effects of MQ on neuronal toxicity and cell viability were investigated over a concentration range of 1-100 µM using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays. The influence of MQ on cellular bioenergetics was examined by measuring cellular ATP levels and from the induction of reactive oxygen species (ROS). An in silico approach was used to assess the potential neurotoxicity of MQ mediated via binding to the active sites of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) and then experimentally validated via in vitro enzymatic assays. MQ was cytotoxic to neuronal cells in a concentration and exposure duration dependent manner and induced a significant reduction in viability at concentrations of ≥25 µM after a 24 h exposure. MQ adversely impacted cellular bioenergetics and significantly depleted ATP production at concentrations of ≥1 µM after 24 h. MQ-induced cellular ROS production, which was correlated with the induction of apoptosis, as revealed by flow cytometry. In silico studies suggested that MQ was a dual cholinesterase inhibitor and one with remarkably potent binding to BuChE. Modelling data were supported by in vitro studies which showed that MQ inhibited both human AChE and BuChE enzymes. In summary, MQ is an antimalarial drug that may induce neurotoxicity by impacting cellular bioenergetics and perturbing the activity of cholinesterases at exposure concentrations relevant to human dosage.
... Cell sample preparation and proteomics analysis KYSE150 cells (1 × 10 6 ) were treated with 10 µM PZM for 24 h, then cells were gathered for protein extraction. Samples preparation and proteomics analysis were executed as described previously [50]. The mass spectrometry proteomics data had been deposited to the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) ...
Targeted therapy attempts are needed to enhance esophageal squamous cell carcinoma (ESCC) patients’ overall survival and satisfaction of life. Nuclear factor erythroid 2-related factor 2 (NRF2), as a high-confidence cancer driver gene, controls the antioxidant response, metabolic balance and redox homeostasis in cancer and is regarded as a potent molecular target for cancer treatment. Here, we attempted to find a new NRF2 inhibitor and study the underlying molecular mechanism in ESCC. We found that up-regulated NRF2 protein was negatively correlated with patient prognosis and promoted tumor proliferation in ESCC. Moreover, Pizotifen malate (PZM), a FDA-approved medication, bound to the Neh1 domain of NRF2 and prevented NRF2 protein binding to the ARE motif of target genes, suppressing transcription activity of NRF2. PZM treatment suppressed tumor development in ESCC PDX model by inducing ferroptosis via down-regulating the transcription of GPX4, GCLC, ME1 and G6PD. Our study illustrates that the over expression of NRF2 indicates poor prognosis and promotes tumor proliferation in ESCC. PZM, as a novel NRF2 inhibitor, inhibits the tumor growth by inducing ferroptosis and elucidates a potent NRF2-based therapy strategy for patients with ESCC.
... For example, it has been found that autophagy can promote the proliferation and invasion of tumor cells and is recognized as a tumor promoter in ESCC (34). Furthermore, Xie et al (35) testified that the induction of mitochondrial autophagy could inhibit the tumor growth of ESCC treated with mefloquine. Therefore, the expression of autophagy-related proteins was also measured herein. ...
Esophageal squamous cell carcinoma (ESCC) is a deadly disease that seriously affects global public health. The aim of the present study was to explore the role of integrin subunit α3 (ITGA3) in ESCC and investigate its detailed molecular mechanisms. Using reverse transcription-quantitative PCR (RT-qPCR) and western blotting, the mRNA and protein expression of ITGA3 in cell lines was detected. In addition, a series of cellular biological experiments, including Cell Counting Kit-8, wound-healing, Transwell and TUNEL assays, were used to evaluate proliferation, migration, invasion and apoptosis, respectively. Furthermore, western blotting was used to measure the expression of corresponding proteins. ITGA3 was found to be upregulated in ESCC cell lines (ECA109 and TE1). It was also found that ITGA3 silencing inhibited the proliferation, migration, invasion and autophagy of ECA109 and TE1 cells but promoted their apoptosis. In addition, ITGA3 silencing was found to inhibit the FAK/PI3K/AKT signaling pathway. In conclusion, ITGA3 knockdown suppressed cell proliferation, invasion, migration and autophagy in ECA109 and TE1 cells, suggesting that ITGA3 may be a potential therapeutic target for the treatment of ESCC.
... The cells were cytogenetically tested by STR-Promega and authenticated. Cells were cultured as previously described [52]. F The inhibitory effect of oxethazaine on AURKA knockout cells was detected by proliferation assay after 72 h. ...
... The anchorage-independent cell growth assay was performed as described previously [52]. Cells were seeded into six-well plates with 8000 cells per well with or without oxethazaine treatment. ...
... The apoptosis assay was performed as described previously [52]. Cells were seeded into 60 mm dishes (2 × 10 5 cells per dish) and incubated with or without oxethazaine treatment. ...
Esophageal squamous cell carcinoma (ESCC), a malignant neoplasm with high incidence, is a severe global public health threat. The current modalities used for treating ESCC include surgery, chemotherapy, and radiotherapy. Although ESCC management and treatment strategies have improved over the last decade, the overall 5-year survival rate remains <20%. Therefore, the identification of novel therapeutic strategies that can increase ESCC patient survival rates is urgently needed. Oxethazaine, an amino-amide anesthetic agent, is mainly prescribed in combination with antacids to relieve esophagitis, dyspepsia, and other gastric disorders. In the present study, we found that oxethazaine inhibited the proliferation and migration of esophageal cancer cells. According to the results of in vitro screening and binding assays, oxethazaine binds directly to AURKA, suppresses AURKA activity, and inhibits the downstream effectors of AURKA. Notably, we found that oxethazaine suppressed tumor growth in three patient-derived esophageal xenograft mouse models and tumor metastasis in vivo. Our findings suggest that oxethazaine can inhibit ESCC proliferation and metastasis in vitro and in vivo by targeting AURKA.
... Interestingly, these cells show activation rather than inhibition of the lysosome by mefloquine. Similarly, another report suggests that mefloquine accelerates mitophagy (Xie et al., 2020). This finding is supported by the observation of increased colocalization of LC3-II and mitochondrial proteins (such as MTCO1) in mefloquine-treated esophageal cancer cells. ...
... Mefloquine is a potential inducer of mitochondrial dysfunction. Mefloquine not only alters the essential levels of mitochondrial proteins (such as the CI subunit NDUFB8, CII-30 kDa, CIII core protein 2, CIV subunit I, and CV alpha subunit) involved in oxidative phosphorylation, but also shifts the NAD + /NADH ratio, leading to oxidative stress and structural changes in mitochondria (Xie et al., 2020). The reduction in oxidative phosphorylation in mitochondria is due to mefloquine targeting the F 0 F 1 proton ATPase complex (Martin-Galiano et al., 2002). ...
Mefloquine is a quinoline-based compound widely used as an antimalarial drug, particularly in chemoprophylaxis. Although decades of research have identified various aspects of mefloquine's anti-Plasmodium properties, toxic effects offset its robust use in humans. Mefloquine exerts harmful effects in several types of human cells by targeting many of the cellular lipids, proteins, and complexes, thereby blocking a number of downstream signaling cascades. In general, mefloquine modulates several cellular phenomena, such as alteration of membrane potential, induction of oxidative stress, imbalance of ion homeostasis, disruption of metabolism, failure of organelle function, etc., leading to cell cycle arrest and programmed cell death. This review aims to summarize the information on functional and mechanistic findings related to the cytotoxic effects of mefloquine.
... Not only that, but there is a strong interest in reusing metformin to prevent cancer and cancer recurrence (22). Our recently study indicated atorvastatin, a lipid-lowering drug (23), and mefloquine, an anti-malarial drug (24), had a strong inhibitory effects on ESCC proliferation. Here, we screened the FDAapproved drug and found tegaserod maleate could inhibit proliferation of ESCC both in vivo and in vitro. ...
Esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC) are the two major types of esophageal cancer (EC). ESCC accounts for 90% of EC. Recurrence after primary treatment is the main reason for poor survival. Therefore, recurrence prevention is a promising strategy for extending the 5-year survival rate. Here, we found tegaserod maleate could inhibit ESCC proliferation both in vivo and in vitro. Proteomics analysis revealed that tegaserod maleate suppressed the peroxisome signaling pathway, in which the key molecules peroxisome membrane protein 11B (PEX11B) and peroxisome membrane protein 13 (PEX13) were downregulated. The immunofluorescence, catalase activity assay, and reactive oxygen species (ROS) confirmed that downregulation of these proteins was related to impaired peroxisome function. Furthermore, we found that PEX11B and PEX13 were highly expressed in ESCC, and knockout of PEX11B and PEX13 further demonstrated the antitumor effect of tegaserod maleate. Importantly, tegaserod maleate repressed ESCC tumor growth in a patient-derived xenograft (PDX) model in vivo. Our findings conclusively demonstrated that tegaserod maleate inhibits the proliferation of ESCC by suppressing the peroxisome pathway.
... 67 Interestingly, Rebecca et al demonstrated a different autophagy-inducing mechanism of CQ derivatives. 68,69 They identified palmitoyl-protein thioesterase 1 (PPT1) as a common target of CQ derivatives including hydroxychloroquine (HCQ), DC661, and Lys05. 68,69 PPT1 functions as a thioester bond breaker of the palmitoylated proteins. ...
Drug repurposing is a feasible strategy in developing novel medications. Regarding the cancer field, scientists are continuously making efforts to redirect conventional drugs into cancer treatment. This approach aims at exploring new applications in the existing agents. Antiparasitic medications, including artemisinin derivatives (ARTs), quinine-related compounds, niclosamide, ivermectin, albendazole derivatives, nitazoxanide and pyrimethamine, have been deeply investigated and widely applied in treating various parasitic diseases for a long time. Generally, their pharmacokinetic and pharmacodynamic properties are well understood, while the side effects are roughly acceptable. Scientists noticed that some of these agents have anticancer potentials and explored the underlying mechanisms to achieve drug repurposing. Recent studies show that these agents inhibit cancer progression via multiple interesting ways, inducing ferroptosis induction, autophagy regulation, mitochondrial disturbance, immunoregulation, and metabolic disruption. In this review, we summarize the recent advancement in uncovering antiparasitic drugs’ anticancer properties from the perspective of their pharmacological targets. Instead of paying attention to the previously discovered mechanisms, we focus more on newly emerging ones that are worth noticing. While most investigations are focusing on the mechanisms of their antiparasitic effect, more in vivo exploration in clinical trials in the future is necessary. Moreover, we also paid attention to what limits the clinical application of these agents. For some of these agents like ARTs and niclosamide, drug modification, novel delivery system invention, or drug combination are strongly recommended for future exploration.
Esophageal squamous cell carcinoma (ESCC), the most common pathological type of esophageal cancer, is one of the leading causes of cancer death worldwide, but an effective therapeutic avenue is still absent. This study was to evaluate the therapeutic effect of Agaricus blazei Murrill polysaccharides (ABP) and Enteromorpha prolifera polysaccharides (EP) on ESCC. The results showed that both ABP and EP have therapeutic effects on multiple malignant processes of ESCC, such as proliferation, migration, invasion, and apoptosis, in vitro. Moreover, both ABP and EP retard tumor growth in vivo and the growth of ESCC-derived organoids. A series of genes and proteins changed upon treatments with ABP and EP after RNA sequencing and proteomics analysis. Autophagy, mitophagy and ferroptosis were enriched after treatments with these two polysaccharides, suggesting that the consumption of ABP and EP may represent a promising dietary supplement for ESCC therapy.
Background: Mitophagy is a cellular survival mechanism that is indispensable in carcinogenesis and tumor progression. However, the research on the mechanism of mitophagy in esophageal cancer metastasis is limited. This study explored the regulatory mechanism by which RECQL4 regulates mitophagy and affects esophageal cancer metastasis.
Methods: The RECQL4 expression in esophageal cancer tissues was examined by bioinformatics analysis and validated by qRT-PCR. Bioinformatics analysis was used to determine the upstream regulatory factor of RECQL4, CREB1, and find the correlation between them. Their binding relationship was evaluated using dual luciferase assay and Chromatin Immunoprecipitation (ChIP) assay. The role of RECQL4 in the viability and metastasis of esophageal cancer cells was investigated through experiments including CCK-8, Transwell, and immunofluorescence. GFP-LC3 plasmid was transfected into cells, followed by western blot (WB) to detect the expression of autophagy marker proteins LC3Ⅰ, LC3Ⅱ, and p62 to analyze mitophagy.
Results: The expression of RECQL4 was up-regulated in esophageal cancer tissues and cells. Overexpression of RECQL4 promoted the viability, invasion, migration, and epithelial-mesenchymal transition (EMT) of esophageal cancer cells by inhibiting mitophagy. Bioinformatics analysis found a substantial positive correlation between the expression of RECQL4 and its upstream transcription factor, CREB1, followed by validation of their binding relationship using dual luciferase and ChIP assays. CREB1 knockdown promoted mitophagy and prevented the metastasis of cancer cells, which could be countered by overexpressing RECQL4.
Conclusion: In this study, CREB1 was found to transcriptionally activate RECQL4 to inhibit mitophagy, thereby promoting esophageal cancer metastasis. This suggests that targeting mitophagy could be an effective therapeutic approach for esophageal cancer.
