Sitagliptin inhibits the survival, stemness and autophagy of glioma cells, and enhances temozolomide cytotoxicity.

The standard regimen treatment has improved GBM outcomes, but the survival rate of patients is still unsatisfactory. Temozolomide (TMZ) resistance is one of main reasons limiting the therapeutic efficacy of GBM. However, there are currently no TMZ-sensitizing drugs available in the clinic. Here we aimed to study whether the antidiabetic drug Sitagliptin can inhibit the survival, stemness and autophagy of GBM cells, and thus enhance TMZ cytotoxicity. We used CCK-8, EdU, colony formation, TUNEL and flow cytometry assays to assess cell proliferation and apoptosis; sphere formation and limiting dilution assays to measure self-renewal and stemness of glioma stem cells (GSCs); Western blot, qRT-PCR or immunohistochemical analysis to measure the expression of proliferation or stem cell markers; Western blot/fluorescent analysis of LC3 and other molecules to evaluate autophagy formation and degradation in glioma cells. We found that Sitagliptin inhibited proliferation and induced apoptosis in GBM cells and suppressed self-renewal and stemness of GSCs. The in vitro findings were further confirmed in glioma intracranial xenograft models. Sitagliptin administration prolonged the survival time of tumor-bearing mice. Sitagliptin could inhibit TMZ-induced protective autophagy and enhance the cytotoxicity of TMZ in glioma cells. In addition, Sitagliptin acted as a dipeptidyl peptidase 4 inhibitor in glioma as well as in diabetes, but it did not affect the blood glucose level and body weight of mice. These findings suggest that Sitagliptin with established pharmacologic and safety profiles could be repurposed as an antiglioma drug to overcome TMZ resistance, providing a new option for GBM therapy.

Body talk on social networking sites and restrained eating among adolescents: A test of a multiple mediation model.

The present study aimed to examine the link between body talk on social networking sites (SNS body talk) and restrained eating as well as the mediating roles of fear of negative appearance evaluation (FNAE) and body shame in this relationship. Furthermore, the moderating role of gender in the mediation model was also examined. A sample of 1481 Chinese middle and high school students (720 girls, mean age = 15.49 ± 1.84) completed self-reported measures of SNS body talk, FNAE, body shame, and restrained eating. Results showed that SNS body talk was positively associated with restrained eating. FNAE and body shame mediated the association between SNS body talk and restrained eating, separately and serially. Furthermore, gender difference emerged in the path from SNS body talk to body shame and the path from FNAE to body shame. Findings of the present study enrich the literature on social media and body image and eating behaviors, and have important implications for reducing negative body image and disordered eating among adolescents.

Structural and functional insights into the ATP-binding cassette transporter family in the corn planthopper, Peregrinus maidis.

The corn planthopper, Peregrinus maidis, is an economically important pest of maize and sorghum. Its feeding behavior and the viruses it transmits can significantly reduce crop yield. The control of P. maidis and its associated viruses relies heavily on insecticides. However, control has proven difficult due to limited direct exposure of P. maidis to insecticides and rapid development of resistance. As such, alternative control methods are needed. In the absence of a genome assembly for this species, we first developed transcriptomic resources. Then, with the goal of finding targets for RNAi-based control, we identified members of the ATP-binding cassette (ABC) transporter family and targeted specific members via RNAi. PmABCB_160306_3, PmABCE_118332_5 and PmABCF_24241_1, whose orthologs in other insects have proven important in development, were selected for knockdown. We found that RNAi-mediated silencing of PmABCB_160306_3 impeded ovary development; disruption of PmABCE_118332_5 resulted in localized melanization; and knockdown of PmABCE_118332_5 or PmABCF_24241_1 each led to high mortality within five days. Each phenotype is similar to that found when targeting the orthologous gene in other species and demonstrates their potential for use in RNAi-based P. maidis control. The transcriptomic data and RNAi results presented here will no doubt assist with the development of new control methods for this pest. This article is protected by copyright. All rights reserved.

Soil microbial responses to large changes in precipitation with nitrogen deposition in an arid ecosystem.

Climatic change severely affects terrestrial ecosystem functioning by modifying soil microbial communities, especially in arid ecosystems. However, how precipitation patterns affect soil microbes and the underlying mechanisms remain largely unclear, particularly under long-term dry-wet cycling and vice versa in field settings. In this study, a field experiment was conducted to quantify soil microbial responses and resilience to precipitation changes with nitrogen addition. We established five levels of precipitation with nitrogen addition over the first 3 years and then balanced this with compensatory precipitation in the fourth year (i.e., reversed the precipitation treatments), to recover to the levels expected over 4 years in a desert steppe ecosystem. Soil microbial community biomass increased with increasing precipitation, and the reversed precipitation reversed these responses. The soil microbial response ratio was constrained by the initial reduction in precipitation, whereas the resilience and limitation/promotion index of most microbial groups tended to increase. Nitrogen addition reduced the response rates of most microbial groups, depending on the soil depth. The soil microbial response and limitation/promotion index could be distinguished by antecedent soil features. The precipitation regime can regulate the responses of soil microbial communities to climatic change via two potential mechanisms: (1) concurrent nitrogen deposition and (2) soil chemical and biological mediation. Soil microbial behaviors and their associations with soil properties should be considered when assessing the responses of terrestrial ecosystems to climatic change. This article is protected by copyright. All rights reserved.

A draft Diabrotica virgifera virgifera genome: insights into control and host plant adaption by a major maize pest insect.

BACKGROUND: Adaptations by arthropod pests to host plant defenses of crops determine their impacts on agricultural production. The larval host range of western corn rootworm, Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), is restricted to maize and a few grasses. Resistance of D. v. virgifera to crop rotation practices and multiple insecticides contributes to its status as the most damaging pest of cultivated maize in North America and Europe. The extent to which adaptations by this pest contributes to host plant specialization remains unknown. RESULTS: A 2.42 Gb draft D. v. virgifera genome, Dvir_v2.0, was assembled from short shotgun reads and scaffolded using long-insert mate-pair, transcriptome and linked read data. K-mer analysis predicted a repeat content of ≥ 61.5%. Ortholog assignments for Dvir_2.0 RefSeq models predict a greater number of species-specific gene duplications, including expansions in ATP binding cassette transporter and chemosensory gene families, than in other Coleoptera. A majority of annotated D. v. virgifera cytochrome P450s belong to CYP4, 6, and 9 clades. A total of 5,404 transcripts were differentially-expressed between D. v. virgifera larvae fed maize roots compared to alternative host (Miscanthus), a marginal host (Panicum virgatum), a poor host (Sorghum bicolor) and starvation treatments; Among differentially-expressed transcripts, 1,908 were shared across treatments and the least number were between Miscanthus compared to maize. Differentially-expressed transcripts were enriched for putative spliceosome, proteosome, and intracellular transport functions. General stress pathway functions were unique and enriched among up-regulated transcripts in marginal host, poor host, and starvation responses compared to responses on primary (maize) and alternate hosts. CONCLUSIONS: Manual annotation of D. v. virgifera Dvir_2.0 RefSeq models predicted expansion of paralogs with gene families putatively involved in insecticide resistance and chemosensory perception. Our study also suggests that adaptations of D. v. virgifera larvae to feeding on an alternate host plant invoke fewer transcriptional changes compared to marginal or poor hosts. The shared up-regulation of stress response pathways between marginal host and poor host, and starvation treatments may reflect nutrient deprivation. This study provides insight into transcriptomic responses of larval feeding on different host plants and resources for genomic research on this economically significant pest of maize.

A Highly Enantioselective Homoenolate Michael Addition/Esterification Sequence of Cyclohexadienone-Tethered Enals via NHC Catalysis.

Reported here is a highly enantioselective homoenolate Michael addition/esterification sequence of cyclohexadienone-tethered enals via N-heterocyclic carbene (NHC) catalysis, affording the enantiopure cis-hydrobenzofurans, cis-hydroindoles, and cis-hydroindenes. The NHC catalyst bearing a nitro group greatly enhances the stereocontrol, and a bulky N-aryl substituent of the triazolium salt in the catalyst is helpful for inhibiting the further aldol condensation after homoenolate Michael addition. The utility of this protocol is highlighted by a gram-scale experiment and versatile downstream transformations.

TRPV4 is not the molecular sensor for bacterial lipopolysaccharides-induced calcium signaling.

Lipopolysaccharides (LPS) is one of the most potent pathogen-associated signals for the immune system of vertebrates. In addition to the canonical pathway of LPS detection mediated by toll-like receptor 4 (TLR4) signaling pathway, TRP channel-mediated pathways endow sensory neurons and epithelial cells with the ability to detect and react to bacterial endotoxins. Previous work revealed that LPS triggers TRPV4-dependent calcium influx in urothelial cells (UCs) and mouse tracheobronchial epithelial cells (mTEC). In marked contrast, here we show that most subtypes of LPS could not directly activate TRPV4 channel. Although LPS from Salmonella enterica serotype Minnesota evoked a [Ca2+]i response in freshly isolated human bronchial epithelial cells (ECs), freshly isolated mouse ear skin single-cell suspensions, or HEK293T cells transiently transfected with mTRPV4, this activation occurred in a TRPV4-independent manner. Additionally, LPS from either E. coli strains or Salmonella enterica serotype Minnesota did not evoke significant difference in inflammation and pain hyperalgesia between wild type and TRPV4 deficient mice. In summary, our results demonstrate that in vitro and in vivo effects induced by LPS are independent of TRPV4, thus providing a clarity to the questioned role of LPS in TRPV4 activation.

Mitochondrion-targeting and in situ photocontrolled protein delivery via photocages.

Defects in mitochondrial proteostasis contribute to many disorders, including cancer, neurodegeneration, and metabolic and genetic diseases. A strategy aimed at restoring the damaged mitochondrial proteostasis is the mitochondrion-targeting and carrier-free delivery of exogenous functional proteins that can replace the endogenous dysfunctional proteins. The modification of a protein with a photolabile protecting group (PPG, i.e., photocage group) can be activated in situ by response to illumination, leading to release of the protein from its photocage. Here, the Cys and peptide photocages with coumarin were first prepared and characterized for proof of concept. Then, we designed a pair of photocage groups PPG-RhB and PPG-TPP using coumarin and mitochondrion-targeting Rhodamine B (RhB) and triphenylphosphine (TPP), and another pair of organelle-nontarget photocage groups Br-PPG and NO2-PPG for comparison. The proteins modified with these two pairs of photocage groups undergo photolysis in solutions, and can penetrate cell membrane toward their destinations in the carrier-free fashions. The intracellular protein photocages are in situ activated by illumination at 405 nm, and the proteins are released from their photocages in mitochondria and cytoplasm, respectively. This strategy of light-responsive and carrier-free cellular delivery enables mitochondrial and cytoplasmic accumulation of exogenous proteins.

Peptide Vaccine Against ADAMTS-7 Ameliorates Atherosclerosis and Postinjury Neointima Hyperplasia.

BACKGROUND: The metalloprotease ADAMTS-7 (a disintegrin and metalloproteinase with thrombospondin type 1 motif 7) is a novel locus associated with human coronary atherosclerosis. ADAMTS-7 deletion protects against atherosclerosis and vascular restenosis in rodents. METHODS: We designed 3 potential vaccines consisting of distinct B cell epitopic peptides derived from ADAMTS-7 and conjugated with the carrier protein KLH (keyhole limpet hemocyanin) as well as aluminum hydroxide as an adjuvant. Arterial ligation or wire injury was used to induce neointima in mice, whereas ApoE-/- and LDLR-/- (LDLR [low-density lipoprotein receptor]) mice fed a high-fat diet were applied to assess atherosclerosis. In addition, coronary stent implantation was performed on vaccine-immunized Bama miniature pigs, followed by optical coherence tomography to evaluate coronary intimal hyperplasia. RESULTS: A vaccine, ATS7vac, was screened out from 3 candidates to effectively inhibit intimal thickening in murine carotid artery ligation models after vaccination. As well, immunization with ATS7vac alleviated neointima formation in murine wire injury models and mitigated atherosclerotic lesions in both hyperlipidemic ApoE-/- and LDLR-/- mice without lowering lipid levels. Preclinically, ATS7vac markedly impeded intimal hyperplasia in swine stented coronary arteries, but without significant immune-related organ injuries. Mechanistically, ATS7vac vaccination produced specific antibodies against ADAMTS-7, which markedly repressed ADAMTS-7-mediated COMP (cartilage oligomeric matrix protein) and TSP-1 (thrombospondin-1) degradation and subsequently inhibited vascular smooth muscle cell migration but promoted re-endothelialization. CONCLUSIONS: ATS7vac is a novel atherosclerosis vaccine that also alleviates in-stent restenosis. The application of ATS7vac would be a complementary therapeutic avenue to the current lipid-lowering strategy for atherosclerotic disease.

Calcium Phosphate Bone Cements Incorporated with Black Phosphorus Nanosheets Enhanced Osteogenesis.

For decades, calcium phosphate bone cements (CPCs) showed impressive advantages for their good biocompatibility, injectability, and osteoconductivity in the bone repair field. However, it is still difficult to prepare CPCs with outstanding antibacterial and self-curing properties, sufficient phosphorus release, and osteoinductivity for clinical application. Herein, we used partially crystallized calcium phosphate and dicalcium phosphate anhydrate particles incorporated with black phosphorous nanosheets to prepare calcium phosphate bone cements (CPCs). The curing time, compressive strength, photothermal properties, and degradation performance of BP/CPC were investigated. In addition, the cytocompatibility and osteoinductivity of BP/CPC were evaluated by cell adhesion, cytotoxicity, alkaline phosphatase detection, alizarin red staining, and western blot assay. The results indicated that BP/CPC showed adjustable curing time, good cytocompatibility, outstanding photothermal properties, and osteoinductivity, suggesting their potential application for bone regeneration.

Hyaluronic acid mediated Fe3O4 nanocubes reversing the EMT through targeted cancer stem cell.

Hepatocellular carcinoma (HCC) is one of the deadliest tumors in the world with a high rate of recurrence and metastasis. Therefore, the most pressing issue today is the development of new drugs, diagnostic and therapeutic approaches for effective cancer treatment. Cancer stem cells (CSCs) play a pivotal role in tumor recurrence, tumor resistance, and tumor metastasis, which provides a new perspective on the development of liver cancer. In the study, a high-temperature thermal breakdown approach was used to create composite magnetic nanocubes modified by polyethyleneimine (PEI) and hyaluronic acid (HA). The Fe3O4 nanocubes can recognize HCC stem cells via receptor-ligand binding of HA and CD44 (HA receptor). While loading a small molecule LDN193189 inhibited the expression of stemness-related genes OCT4 and Nanog. More crucially, the Fe3O4 nanocubes significantly suppressed HCC cell proliferation and migration by regulating the expression of epithelial-mesenchymal transition (EMT) process markers E-cadherin, Vimentin, and N-cadherin. Dual targeting using magnetic and receptor-mediated targeting improved the uptake of the drug delivery system. Our findings imply that the medication delivery method might be a potential therapeutic strategy for HCC.

The influence of arts engagement on the mental health of isolated college students during the COVID-19 outbreak in China.

Objectives: The COVID-19 pandemic significantly impacted the mental health of college students. This study aimed to investigate the buffering effect of arts engagement on anxiety and resilience in college students during the COVID-19 pandemic. Study design: A cross-sectional study. Methods: The data were collected via an online survey during a wave of SARS-CoV-2 infections in Shanghai (March 15 to April 15, 2022). In total, 2,453 college students throughout China reported general anxiety symptom levels (according to the GAD-7), resilience (according to the Connor-Davidson Resilience Scale), frequency of receptive arts engagement in the previous year, exposure to risk situations, and behavioral changes due to the pandemic. Results: During the current stage of the pandemic, 43.7% of college students suffered from varying degrees of anxiety, and 2.6% showed severe anxiety. Gender and learning stage were not associated with anxiety. Hierarchical regression analysis showed that the decision to return to academic institution, the degree of exposure to COVID-19, and the frequency of accepting art participation and resilience could significantly predict the anxiety level of college students. Gender, study stage, behavioral changes arising from COVID-19, and exposure to COVID-19 significantly predict the resilience level of college students. There was an association between high frequency music activities, reading activities and low anxiety level (p < 0.001). There was an association between high frequency digital art, music activities, reading and high resilience (p < 0.01). Conclusions: Arts engagement appears to help students cope with mental health problems and those at risk. Policymakers should encourage college students to participate in art activities, especially in the context of social distancing.

Peer Appearance Teasing and Restrained Eating Among Chinese Adolescent Girls: A Mediation Model of Fear of Negative Appearance Evaluation and Body Surveillance.

Restrained eating is becoming widespread recently. The present study aimed to examine whether peer appearance teasing was positively associated with restrained eating among Chinese adolescent girls and whether this association was mediated by fear of negative appearance evaluation (FNAE) and body surveillance. A sample of 720 Chinese adolescent girls completed questionnaires on peer appearance teasing, FNAE, body surveillance, and restrained eating. Peer appearance teasing was significantly and positively related to girls' restrained eating. FNAE mediated the relationship between peer appearance teasing and restrained eating. Furthermore, the association between peer appearance teasing and restrained eating was sequentially mediated by FNAE and body surveillance. The findings of this study provide new insights into the development of restrained eating among adolescent girls, and highlight peer appearance teasing and body surveillance as potential targets for intervention programs regarding body image concerns and eating disorders.

Compensatory growth as a response to post-drought in grassland.

Grasslands are structurally and functionally controlled by water availability. Ongoing global change is threatening the sustainability of grassland ecosystems through chronic alterations in climate patterns and resource availability, as well as by the increasing frequency and intensity of anthropogenic perturbations. Compared with many studies on how grassland ecosystems respond during drought, there are far fewer studies focused on grassland dynamics after drought. Compensatory growth, as the ability of plants to offset the adverse effects of environmental or anthropogenic perturbations, is a common phenomenon in grassland. However, compensatory growth induced by drought and its underlying mechanism across grasslands remains not clear. In this review, we provide examples of analogous compensatory growth from different grassland types across drought characteristics (intensity, timing, and duration) and explain the effect of resource availability on compensatory growth and their underlying mechanisms. Based on our review of the literature, a hypothetic framework for integrating plant, root, and microbial responses is also proposed to increase our understanding of compensatory growth after drought. This research will advance our understanding of the mechanisms of grassland ecosystem functioning in response to climate change.

DNA Nanoribbon for Efficient Anti-miRNA Peptide Nucleic Acid Delivery and Synergistic Enhancement of Cancer Cell Apoptosis.

Antisense peptide nucleic acid (asPNA), an effective antisense drug, has been employed as a gene therapy agent and a useful tool in molecular biology. Gaining control over the delivery of asPNA to target tissues has been a major hindrance to its wide application in clinical practice. A simple and efficient DNA nanoribbon (DNR)-based drug delivery process has been designed in this study that releases the asPNA agent to inhibit oncogenic microRNAs (miRNAs). Furthermore, we demonstrated how the AS1411 aptamer that binds nucleolin on the cell membranes works as a control mechanism capable of identifying target cancer cells and enhancing the enrichment capacity of DNR. With the biodegradability of DNR, we can efficiently initiate the release of asPNA into the cytoplasm, particularly targeting the intended miR-21 and synergistically increasing programmed cell death 4 (PDCD4) expression to enhance cell apoptosis. We assume that this well-defined delivery mechanism will aid in designing antisense site-specific treatments for various diseases, including cancer.

Efficacy of periodontal soft tissue augmentation prior to orthodontic treatment on preventing gingival recession: study protocol for a randomised controlled trial.

INTRODUCTION: In adult patients with thin phenotype, gingival recession is obvious in the mandibular incisors after a large amount of lingual tooth movement. A systematic review indicated that soft tissue augmentation before orthodontic treatment might yield satisfactory results with respect to the progression of gingival recession. However, the studies included had a low-to-moderate level of evidence. This study was designed to investigate the efficacy of soft tissue augmentation prior to orthodontic treatment on the prevention of gingival recession. METHOD AND ANALYSIS: This is a single-centre, single-blind, randomised controlled, double-armed parallel group comparison trial. This study was initiated in December 2021 and end in December 2025 (anticipated). Patients with mild crowding in the lower arch and a thin gingival type was enrolled and randomly assigned in a 1:1 ratio to either group A (having soft tissue augmentation prior to orthodontic treatment) or group B (having orthodontic treatment only). The planned number of enrolled patients was 48 (24 patients × 2 groups). The primary endpoint was the mean change in recession of the gingival margin. Secondary endpoints included the probing depth, gingival phenotype, frequency of complete root coverage, gingival thickness, width of the keratinised gingiva, clinical attachment level, gingival recession class, full-mouth plaque score, alveolar bone thickness in the faciolingual dimension of the lower anterior teeth and lower incisor inclination. ETHICS AND DISSEMINATION: The study protocol was approved by the Ethical Committee of the Shanghai Stomatological Hospital (certificate number (2021)016). The study was conducted in accordance with the Declaration of Helsinki, the Clinical Trials Act and other current legal regulations in China. Written informed consent was obtained from all the participants. The results of this study will be reported in journal publications. TRIAL REGISTRATION NUMBER: ChiCTR2100050892.

[Retracted] miR­29b enhances the proliferation and migration of bone marrow mesenchymal stem cells in rats with castration­induced osteoporosis through the PI3K/AKT and TGF­ß/Smad signaling pathways.

[This retracts the article DOI: 10.3892/etm.2020.9045.].

"I wanna look like the person in that picture": Linking selfies on social media to cosmetic surgery consideration based on the tripartite influence model.

Based on the tripartite influence model, the present study linked investment in others' selfies on social media to facial dissatisfaction and cosmetic surgery consideration. We also tested the mediating role of appearance comparisons and general attractiveness internalization in these relationships. Additionally, potential gender differences in the proposed model were examined. A sample of 884 Chinese adolescents (mean age = 16.80 ± 0.92, ranging from 14 to 19 years old) completed self-reported measures of investment in others' selfies, appearance comparisons, general attractiveness internalization, facial dissatisfaction, and cosmetic surgery consideration. Results showed that investment in others' selfies was positively associated with facial dissatisfaction and cosmetic surgery consideration. Path analysis further revealed that investment in others' selfies was associated with appearance comparisons and general attractiveness internalization, which in turn were associated with facial dissatisfaction, and then facial dissatisfaction was related to cosmetic surgery consideration. Additionally, there were indirect associations between investment in others' selfies and cosmetic surgery consideration via general attractiveness internalization and facial dissatisfaction respectively, but not via appearance comparisons. No gender difference was found in the proposed model. Findings of the present study provide preliminary support for the tripartite influence model in the context of investment in others' selfies on social media and cosmetic surgery consideration and enrich the literature on body image and cosmetic surgery in the Chinese context.

Effects of different boiling processes on chemical compositions of Lilii Bulbus soup.

Lilii Bulbus, an edible Chinese herbal medicine, has a long history in medicine. However, research on effectively boiling Lilii Bulbus is rare. To make the more nutritious Lilii Bulbus soup, the optimized boiling process, using an alternate heating mode by decoction pot carrying a mixture of water and Chinese liquor at the ration of 9:1, was established in this study. Compared to the soup prepared by the daily process, the polysaccharide amount improved by 54%, and the total heavy metals decreased by 33.5% using the optimized boiling process. In addition, the total saponins at 34.3 µg/g were determined in the soup prepared by the optimized process. Meanwhile, the colchicine content in the boiled Lilii Bulbus soup was undetectable using the optimized process. This research performs an optimized boiling process for making Lilii Bulbus soup, and provides a reference for generating high commercial value from Lilii Bulbus soup in the future.

The NRF2 antagonist ML385 inhibits PI3K-mTOR signaling and growth of lung squamous cell carcinoma cells.

BACKGROUND: Lung squamous cell carcinoma (LUSC) currently has limited therapeutic options because of the relatively few validated targets and the lack of clinical drugs for some of these targets. Although NRF2/NFE2L2 pathway activation commonly occurs in LUSC, NRF2 has predominantly been studied in other cancer models. Here, we investigated the function of NRF2 in LUSC, including in organoid models, and we explored the activity of a small molecule NRF2 inhibitor ML385, which has not previously been investigated in LUSC. METHODS: We first explored the role of NRF2 signaling in LUSC cancer cell line and organoid proliferation through NRF2 knockdown or ML385 treatment, both in vivo and in vitro. Next, we performed Western blot and immunofluorescence assays to determine the effect of NRF2 inhibition on PI3K-mTOR signaling. Finally, we used cell viability and clonogenic assays to explore whether ML385 could sensitize LUSC cancer cells to PI3K inhibitors. RESULTS: We find that downregulation of NRF2 signaling inhibited proliferation of LUSC cancer cell lines and organoids, both in vivo and in vitro. We also demonstrate that inhibition of NRF2 reduces PI3K-mTOR signaling, with two potential mechanisms being involved. Although NRF2 promotes AKT phosphorylation, it also acts downstream of AKT to increase RagD protein expression and recruitment of mTOR to lysosomes after amino acid stimulation. We also find that ML385 potentiates LUSC growth inhibition by a pan-PI3K inhibitor, which correlates with stronger inhibition of PI3K-mTOR signaling. CONCLUSIONS: Our data provide additional support for NRF2 promoting LUSC growth through PI3K-mTOR activation and support development of NRF2 inhibitors for the treatment of LUSC.