Soil phosphorus cycling across a 100-year deforestation chronosequence in the Amazon rainforest.

Deforestation of tropical rainforests is a major land use change that alters terrestrial biogeochemical cycling at local to global scales. Deforestation and subsequent reforestation are likely to impact soil phosphorus (P) cycling, which in P-limited ecosystems such as the Amazon basin has implications for long-term productivity. We used a 100-year replicated observational chronosequence of primary forest conversion to pasture, as well as a 13-year-old secondary forest, to test land use change and duration effects on soil P dynamics in the Amazon basin. By combining sequential extraction and P K-edge X-ray absorption near edge structure (XANES) spectroscopy with soil phosphatase activity assays, we assessed pools and process rates of P cycling in surface soils (0-10 cm depth). Deforestation caused increases in total P (135-398 mg kg-1 ), total organic P (Po ) (19-168 mg kg-1 ), and total inorganic P (Pi ) (30-113 mg kg-1 ) fractions in surface soils with pasture age, with concomitant increases in Pi fractions corroborated by sequential fractionation and XANES spectroscopy. Soil non-labile Po (10-148 mg kg-1 ) increased disproportionately compared to labile Po (from 4-5 to 7-13 mg kg-1 ). Soil phosphomonoesterase and phosphodiesterase binding affinity (Km ) decreased while the specificity constant (Ka ) increased by 83%-159% in 39-100y pastures. Soil P pools and process rates reverted to magnitudes similar to primary forests within 13 years of pasture abandonment. However, the relatively short but representative pre-abandonment pasture duration of our secondary forest may not have entailed significant deforestation effects on soil P cycling, highlighting the need to consider both pasture duration and reforestation age in evaluations of Amazon land use legacies. Although the space-for-time substitution design can entail variation in the initial soil P pools due to atmospheric P deposition, soil properties, and/or primary forest growth, the trend of P pools and process rates with pasture age still provides valuable insights.

Missing phosphorus legacy of the Anthropocene: Quantifying residual phosphorus in the biosphere.

A defining feature of the Anthropocene is the distortion of the biosphere phosphorus (P) cycle. A relatively sudden acceleration of input fluxes without a concomitant increase in output fluxes has led to net accumulation of P in the terrestrial-aquatic continuum. Over the past century, P has been mined from geological deposits to produce crop fertilizers. When P inputs are not fully removed with harvest of crop biomass, the remaining P accumulates in soils. This residual P is a uniquely anthropogenic pool of P, and its management is critical for agronomic and environmental sustainability. Managing residual P first requires its quantification-but measuring residual P is challenging. In this review, we synthesize approaches to quantifying residual P, with emphasis on advantages, disadvantages, and complementarity. Common approaches to estimate residual P are mass balances, long-term experiments, soil test P trends and chronosequences, with varying suitability or even limitations to distinct spatiotemporal scales. We demonstrate that individual quantification approaches are (i) constrained, (ii) often complementary, and (iii) may be feasible at only certain time-space scales. While some of these challenges are inherent to the quantification approach, in many cases there are surmountable challenges that can be addressed by unifying existing P pool and flux datasets, standardizing and synchronizing data collection on pools and fluxes, and quantifying uncertainty. Though defined as a magnitude, the distribution and speciation of residual P is relatively less understood but shapes its utilization and environmental impacts. The form of residual P will vary by agroecosystem context due to edaphoclimatic-specific transformation of the accumulated P, which has implications for management (e.g., crop usage) and future policies (e.g., lag times in P loading from non-point sources). Quantifying the uncertainty in measuring residual P holds value beyond scientific understanding, as it supports prioritization of monitoring and management resources and inform policy.

Pomiferin targets SERCA, mTOR, and P-gp to induce autophagic cell death in apoptosis-resistant cancer cells, and reverses the MDR phenotype in cisplatin-resistant tumors in vivo.

Drug resistance in cancer has been classified as innate resistance or acquired resistance, which were characterized by apoptotic defects and ABC transporters overexpression respectively. Therefore, to preclude or reverse these resistance mechanisms could be a promising strategy to improve chemotherapeutic outcomes. In this study, a natural product from Osage Orange, pomiferin, was identified as a novel autophagy activator that circumvents innate resistance by triggering autophagic cell death via SERCA inhibition and activation of the CaMKKß-AMPK-mTOR signaling cascade. In addition, pomiferin also directly inhibited the P-gp (MDR1/ABCB1) efflux and reversed acquired resistance by potentiating the accumulation and efficacy of the chemotherapeutic agent, cisplatin. In vivo study demonstrated that pomiferin triggered calcium-mediated tumor suppression and exhibited an anti-metastatic effect in the LLC-1 lung cancer-bearing mouse model. Moreover, as an adjuvant, pomiferin potentiated the anti-tumor effect of the chemotherapeutic agent, cisplatin, in RM-1 drug-resistant prostate cancer-bearing mouse model by specially attenuating ABCB1-mediated drug efflux, but not ABCC5, thereby promoting the accumulation of cisplatin in tumors. Collectively, pomiferin may serve as a novel effective agent for circumventing drug resistance in clinical applications.

Solution 31P NMR Investigation of Inositol Hexakisphosphate Surface Complexes at the Amorphous Aluminum Oxyhydroxide-Water Interface.

Phytate (myo-inositol hexakisphosphate, myo-IHP) is one of the most common organic phosphorus (P) species in soils and sediments which can be mineralized to increase the concentration of dissolved phosphate in pore water. Because of its six phosphate functional groups, functional group-specific adsorption mechanisms in reactive soil minerals become important in predicting solubility. In this study, solution 31P NMR was used to elucidate the functional group-specific adsorption mechanisms of myo-IHP at the amorphous Al hydroxide (AAH)-water interface at pH 6.5 in conjunction with batch adsorption experiments and Zetasizer measurements. The adsorption maximum of myo-IHP with AAH was ∼312.50 mmol kg-1, and the charge reversal effects in IHP-reacted AAH particles suggested the presence of inner-sphere surface species. The upfield shifts of various phosphate groups in the NMR spectra further supported the formation of inner-sphere IHP complexes at the AAH-water interface. When the initial myo-IHP/AAH (mol kg-1) was decreased from 2.5 to 1.25-1.67, P1,3 and P4,6 functional groups were coordinated in addition to P2; P5 became reactive with the ratio being decreased to <0.84, P5. This multifunctional group coordination increased aggregate size. The study showed that the availability of surface sites of adsorbents influenced the functional group-specific myo-IHP adsorption.

SERCA and P-glycoprotein inhibition and ATP depletion are necessary for celastrol-induced autophagic cell death and collateral sensitivity in multidrug-resistant tumor cells.

Multidrug resistance (MDR) represents an obstacle in anti-cancer therapy. MDR is caused by multiple mechanisms, involving ATP-binding cassette (ABC) transporters such as P-glycoprotein (P-gp), which reduces intracellular drug levels to sub-therapeutic concentrations. Therefore, sensitizing agents retaining effectiveness against apoptosis- or drug-resistant cancers are desired for the treatment of MDR cancers. The sarcoplasmic/endoplasmic reticulum Ca2+ ATPase (SERCA) pump is an emerging target to overcome MDR, because of its continuous expression and because the calcium transport function is crucial to the survival of tumor cells. Previous studies showed that SERCA inhibitors exhibit anti-cancer effects in Bax-Bak-deficient, apoptosis-resistant and MDR cancers, whereas specific P-gp inhibitors reverse the MDR phenotype of cancer cells by blocking efflux of chemotherapeutic agents. Here, we unraveled SERCA and P-gp as double targets of the triterpenoid, celastrol to reverse MDR. Celastrol inhibited both SERCA and P-gp to stimulate calcium-mediated autophagy and ATP depletion, thereby induced collateral sensitivity in MDR cancer cells. In vivo studies further confirmed that celastrol suppressed tumor growth and metastasis by SERCA-mediated calcium mobilization. To the best of our knowledge, our findings demonstrate collateral sensitivity in MDR cancer cells by simultaneous inhibition of SERCA and P-gp for the first time.

Transport of imidazolium-based ionic liquids with different anion/cation species in sand/soil columns.

Ionic liquids (ILs) have been widely used as environmentally friendly solvents to replace volatile organic solvents in the chemistry industries. They have a high water solubility and potential risk to organisms in the soil-water environment. At present, most studies focused on the batch sorption of ILs in soil and neglected the investigation of IL transports in soil, which results in a lack of understanding of the structure-dependent mobility of ILs in the environment. Laboratory-scale sand/soil column experiments were performed to study the transport of imidazolium-based ILs, such as [C4mim][OTF], [C4mim][TOS], [C4mim][MeSO3], [C4mim][BF4], [C2mim][BF4], and [C6mim][BF4] including different counteranions and alkyl chain lengths of IL cations. Batch experiments were also carried out to compare the difference of sorption distribution coefficient (Kd) between the batch and column experiments. A one-dimensional convective-dispersive model using CXTFIT code was created based on the measured breakthrough curves (BTCs) to estimate the column transport parameters. For the anion, [BF4-], the Kd of ILs in both batch and column experiments increased with increasing alkyl chain lengths of the IL cation. In batch tests, counteranions showed no influence on the Kd of [C4mim][OTF], [C4mim][TOS], [C4mim][MeSO3], [C4mim][BF4], [C2mim][BF4], and [C6mim][BF4]. However, in column tests, the BTCs of 1-butyl-3-methylimidazolium-based ILs were anion dependent as evidenced by the change of retardation factor (R) for different counteranions. Furthermore, the effects of transport distance (11cm, 15cm, 19cm, and 24cm) on the mobility of ILs were estimated. The longer distances signified an increase in the contact time and more binding sites for ILs and therefore, the smoother shapes of BTCs in column experiments.

The relationship between nomophobia and latent classes of personality.

The phenomenon of nomophobia, defined as the anxiety experienced when a person is without their mobile phone or is unable to use it, has been identified as having serious negative effects on individuals, particularly students. Previous research has explored the relationship between personality traits and nomophobia, but the findings have been inconclusive. The main objective of this study was to classify personality types through latent class analysis and explore the relationship between these personality types and nomophobia. The Chinese version of the Nomophobia Scale and the Chinese brief version of the Big Five Personality Inventory were used in this study to survey 1906 Chinese college students. The results indicated that (1) a four-class model provided the best fit and categorized the personality traits as the overcontrolled class, resilient class, moderate class, and vulnerable class; (2) significant differences were observed between the four personality types and nomophobia, with overcontrolled and resilient personality types consistently scoring significantly lower than moderate and vulnerable personality types. Our finding highlights the key feature of the study.

Onjisaponin B derived from Radix Polygalae enhances autophagy and accelerates the degradation of mutant α-synuclein and huntingtin in PC-12 cells.

Emerging evidence indicates important protective roles being played by autophagy in neurodegenerative disorders through clearance of aggregate-prone or mutant proteins. In the current study, we aimed to identify autophagy inducers from Chinese medicinal herbs as a potential neuroprotective agent that enhances the clearance of mutant huntingtin and α-synuclein in PC-12 cells. Through intensive screening using the green fluorescent protein-light chain 3 (GFP-LC3) autophagy detection platform, we found that the ethanol extracts of Radix Polygalae (Yuan Zhi) were capable of inducing autophagy. Further investigation showed that among three single components derived from Radix Polygalae--i.e., polygalacic acid, senegenin and onjisaponin B--onjisaponin B was able to induce autophagy and accelerate both the removal of mutant huntingtin and A53T α-synuclein, which are highly associated with Huntington disease and Parkinson disease, respectively. Our study further demonstrated that onjisaponin B induces autophagy via the AMPK-mTOR signaling pathway. Therefore, findings in the current study provide detailed insights into the protective mechanism of a novel autophagy inducer, which is valuable for further investigation as a new candidate agent for modulating neurodegenerative disorders through the reduction of toxicity and clearance of mutant proteins in the cellular level.

Adsorption mechanisms of inositol hexakisphosphate in the presence of phosphate at the amorphous aluminum oxyhydroxide-water interface.

Myo-inositol hexakisphosphate (myo-IHP) is one of the most common soil organic phosphorus (P) species in soil. Its retention in soil is often competed by phosphate, making bioavailability of P species difficult. In this study, the adsorption mechanism of myo-IHP at the amorphous aluminum (oxyhydr)oxide (AAH)-water interface was investigated at pH 6.5 in the presence of phosphate using batch adsorption experiments and solution 31P NMR spectroscopy. The ratio of [myo-IHP]i/[phosphate]i (Ri) was kept 0.33-3 while ligand addition was varied. In the absence of phosphate, myo-IHP forms inner-sphere surface complexes in AAH via P1,3, P2, P4,6, and P5 functional group coordination. When two ligands were simultaneously added, fewer P functional groups of myo-IHP coordinated to AAH and the surface complexes were altered with the coordination of mainly P1,3 and P2 functional groups. When phosphate was pre-adsorbed, myo-IHP adsorption decreased by 8.0-44% compared to the respective simultaneous addition system. P2 or P5 functional group was predominantly coordinated to the AAH surfaces at Ri = 0.33. Myo-IHP pre-adsorption resulted in an increase in the final myo-IHP adsorption compared to that in the simultaneous addition system under the respective Ri values (0.33-3). In this system, P1,3, P2, P4,6, and P5 functional groups were coordinated to form inner-sphere surface complexes regardless of Ri. The study revealed that the functional group specific adsorption mechanism of myo-IHP at the AAH-water interface was affected by addition sequence and Ri of two ligands. The competitive adsorption between organic P and phosphate plays an important role in the fate of P in soils.

Accumulation and release of organic phosphorus (P) from legacy P-affected soils to adjacent drainage water.

Legacy effects of P in agricultural soils have been highlighted in recent literature. However, co-accumulation and release of organic P (Po) have often been ignored in current agro-environmental assessments. The mineralizable Po fraction has a potential to increase the activity of phosphate in pore water, increasing fertility or degrading water quality. In this study, the effects of agricultural management practices (fertilizer applied corn-soybean rotation cropland and dairy manure applied pasture) on the Po/phosphate ratio were investigated in P-rich (290-1232 mg kg-1) agricultural soils and adjacent ditchwater using experimental soil-water chemistry. The effect of agricultural management was significant on both Po and the Po/phosphate ratio in soil and adjacent ditchwater. The Po content, dominated by orthophosphate monoesters, in the manure-amended pasture (average ~ 245 mg kg-1) was significantly greater than that in the fertilizer-applied cropland (average 103 mg kg-1). The Po/phosphate ratio was also significantly greater in the manure-amended pasture (0.54) than in the fertilizer-applied cropland (0.42). Similarly, water quality data also showed that ditchwater near the pasture had a significantly greater flux of dissolved non-reactive P and a greater Po/phosphate ratio compared to the water near the fertilizer-applied sites. Furthermore, a greater Po/phosphate ratio in ditchwater was often observed during wet periods, and the ratio was positively correlated to the discharge (r = 0.42, p = 0.003). The study showed the agricultural management-specific Po accumulation and release and - Po/phosphate ratio that might affect the fate of P in agroecosystems.

Depth sequence distribution of water extractable colloidal phosphorus and its phosphorus speciation in intensively managed agricultural soils.

Legacy phosphorus (P) has accelerated the subsurface transport of colloidal P (CP) in intensively managed agricultural soils in the Midwestern U.S. Because of its high P sorption capacity and mobility, understanding the depth sequence distribution of mobile CP and its speciation in the soil profile is critical in assessing total P (TP) loss to protect the water quality of adjacent water bodies. In this study, physicochemical properties of water-extractable colloids (WECs) from the soil profile at 0-180 cm were characterized using conventional wet chemical analysis. Solution P-31 nuclear magnetic resonance spectroscopy (NMR), P and Fe K-edge X-ray absorption spectroscopy, and transmission electron microscopy were also used to understand P speciation and mineralogy of CP. Percent recovery of WECs per bulk soil increased more than three times with increasing depth. Considering mildly alkaline pH of pore water and negative zeta potential (-21 ± 4 mV) of WECs (size: 1.65 ± 0.45 µm), the transport of P rich WECs (TP: approximately 210-700 mg kg-1) were facilitated from surface to subsoils. Generally, TP in WEC decreased with increasing depth. Interestingly, WECs in subsoil contain organic P (OP) as much as 60 mg kg-1. NMR analysis clearly showed the presence of OP monoesters, OP diesters, and orthophosphate in these particles. Both orthophosphate and OP species interacted with iron oxyhydroxides, calcite, and aluminol functional groups of gibbsite and or phyllosilicates. The study showed the availability of WECs from surface to subsoils that carry orthophosphate as well as OP in legacy P impacted agricultural soils in the Midwestern U.S.

High flow event induced the subsurface transport of particulate phosphorus and its speciation in agricultural tile drainage system.

Subsurface storm flow of phosphorus (P), including particulate P, has been recently discussed as an important P transport path in contrast to typical surface runoff events. However, P speciation, and P concentration during storm events has not been extensively investigated; therefore, its contribution to the water quality is not clearly understood. In this study, the physicochemical properties of particulate P in tile water samples during a high flow event were investigated in Midwestern agricultural lands using wet chemical methods, 31P Nuclear Magnetic Resonance spectroscopy and P K-edge X-ray absorptions near edge structure spectroscopy. In slightly alkaline pH tile water, total P was ranging from ∼0.06 to 0.22 mg L-1, which is significantly greater than dissolved reactive P (DRP) (∼0.02-0.08 mg L-1). The tile water contains P enriched particulate matters (∼200-660 mg L-1). Total P in the colloidal fraction was from 1013 to 2270 mg kg-1. Phosphate and organic P species, especially monoesters, are sorbed in soil colloids like calcite, and iron oxides, and colloids are effective carriers of P in the subsurface transport process during storm events. The results of this study show that storm events can accelerate the subsurface transport of P with soil particles in addition to DRP.

Neferine induces autophagy-dependent cell death in apoptosis-resistant cancers via ryanodine receptor and Ca2+-dependent mechanism.

Resistance of cancer cells to chemotherapy is a significant clinical concern and mechanisms regulating cell death in cancer therapy, including apoptosis, autophagy or necrosis, have been extensively investigated over the last decade. Accordingly, the identification of medicinal compounds against chemoresistant cancer cells via new mechanism of action is highly desired. Autophagy is important in inducing cell death or survival in cancer therapy. Recently, novel autophagy activators isolated from natural products were shown to induce autophagic cell death in apoptosis-resistant cancer cells in a calcium-dependent manner. Therefore, enhancement of autophagy may serve as additional therapeutic strategy against these resistant cancers. By computational docking analysis, biochemical assays, and advanced live-cell imaging, we identified that neferine, a natural alkaloid from Nelumbo nucifera, induces autophagy by activating the ryanodine receptor and calcium release. With well-known apoptotic agents, such as staurosporine, taxol, doxorubicin, cisplatin and etoposide, utilized as controls, neferine was shown to induce autophagic cell death in a panel of cancer cells, including apoptosis-defective and -resistant cancer cells or isogenic cancer cells, via calcium mobilization through the activation of ryanodine receptor and Ulk-1-PERK and AMPK-mTOR signaling cascades. Taken together, this study provides insights into the cytotoxic mechanism of neferine-induced autophagy through ryanodine receptor activation in resistant cancers.

Ca2+ signalling plays a role in celastrol-mediated suppression of synovial fibroblasts of rheumatoid arthritis patients and experimental arthritis in rats.

BACKGROUND AND PURPOSE: Celastrol exhibits anti-arthritic effects in rheumatoid arthritis (RA), but the role of celastrol-mediated Ca2+ mobilization in treatment of RA remains undefined. Here, we describe a regulatory role for celastrol-induced Ca2+ signalling in synovial fibroblasts of RA patients and adjuvant-induced arthritis (AIA) in rats. EXPERIMENTAL APPROACH: We used computational docking, Ca2+ dynamics and functional assays to study the sarcoplasmic/endoplasmic reticulum Ca2+ ATPase pump (SERCA). In rheumatoid arthritis synovial fibroblasts (RASFs)/rheumatoid arthritis fibroblast-like synoviocytes (RAFLS), mechanisms of Ca2+ -mediated autophagy were analysed by histological, immunohistochemical and flow cytometric techniques. Anti-arthritic effects of celastrol, autophagy induction, and growth rate of synovial fibroblasts in AIA rats were monitored by microCT and immunofluorescence staining. mRNA from joint tissues of AIA rats was isolated for transcriptional analysis of inflammatory genes, using siRNA methods to study calmodulin, calpains, and calcineurin. KEY RESULTS: Celastrol inhibited SERCA to induce autophagy-dependent cytotoxicity in RASFs/RAFLS via Ca2+ /calmodulin-dependent kinase kinase-ß-AMP-activated protein kinase-mTOR pathway and repressed arthritis symptoms in AIA rats. BAPTA/AM hampered the in vitro and in vivo effectiveness of celastrol. Inflammatory- and autoimmunity-associated genes down-regulated by celastrol in joint tissues of AIA rat were restored by BAPTA/AM. Knockdown of calmodulin, calpains, and calcineurin in RAFLS confirmed the role of Ca2+ in celastrol-regulated gene expression. CONCLUSION AND IMPLICATIONS: Celastrol triggered Ca2+ signalling to induce autophagic cell death in RASFs/RAFLS and ameliorated arthritis in AIA rats mediated by calcium-dependent/-binding proteins facilitating the exploitation of anti-arthritic drugs based on manipulation of Ca2+ signalling.

Mode of Action Analyses of Neferine, a Bisbenzylisoquinoline Alkaloid of Lotus (Nelumbo nucifera) against Multidrug-Resistant Tumor Cells.

Neferine, a bisbenzylisoquinoline alkaloid isolated from the green seed embryos of Lotus (Nelumbo nucifera Gaertn), has been previously shown to have various anti-cancer effects. In the present study, we evaluated the effect of neferine in terms of P-glycoprotein (P-gp) inhibition via in vitro cytotoxicity assays, R123 uptake assays in drug-resistant cancer cells, in silico molecular docking analysis on human P-gp and in silico absorption, distribution, metabolism, and excretion (ADME), quantitative structure activity relationships (QSAR) and toxicity analyses. Lipinski rule of five were mainly considered for the ADME evaluation and the preset descriptors including number of hydrogen bond donor, acceptor, hERG IC50, logp, logD were considered for the QSAR analyses. Neferine revealed higher toxicity toward paclitaxel- and doxorubicin-resistant breast, lung or colon cancer cells, implying collateral sensitivity of these cells toward neferine. Increased R123 uptake was observed in a comparable manner to the control P-gp inhibitor, verapamil. Molecular docking analyses revealed that neferine still interacts with P-gp, even if R123 was pre-bound. Bioinformatical ADME and toxicity analyses revealed that neferine possesses the druggability parameters with no predicted toxicity. In conclusion, neferine may allocate the P-gp drug-binding pocket and prevent R123 binding in agreement with P-gp inhibition experiments, where neferine increased R123 uptake.

N-Desmethyldauricine Induces Autophagic Cell Death in Apoptosis-Defective Cells via Ca2+ Mobilization.

Resistance of cancer cells to chemotherapy remains a significant problem in oncology. Mechanisms regulating programmed cell death, including apoptosis, autophagy or necrosis, in the treatment of cancers have been extensively investigated over the last few decades. Autophagy is now emerging as an important pathway in regulating cell death or survival in cancer therapy. Recent studies demonstrated variety of natural small-molecules could induce autophagic cell death in apoptosis-resistant cancer cells, therefore, discovery of novel autophagic enhancers from natural products could be a promising strategy for treatment of chemotherapy-resistant cancer. By computational virtual docking analysis, biochemical assays, and advanced live-cell imaging techniques, we have identified N-desmethyldauricine (LP-4), isolated from rhizoma of Menispermum dauricum DC as a novel inducer of autophagy. LP-4 was shown to induce autophagy via the Ulk-1-PERK and Ca2+/Calmodulin-dependent protein kinase kinase ß (CaMKKß)-AMPK-mTOR signaling cascades, via mobilizing calcium release through inhibition of SERCA, and importantly, lead to autophagic cell death in a panel of cancer cells, apoptosis-defective and apoptosis-resistant cells. Taken together, this study provides detailed insights into the cytotoxic mechanism of a novel autophagic compound that targeting the apoptosis resistant cancer cells, and new implication on drug discovery from natural products for drug resistant cancer therapy.

Autophagic degradation of epidermal growth factor receptor in gefitinib-resistant lung cancer by celastrol.

Drug resistance of non-small cell lung cancer (NSCLC) is highly correlated to the mutation of the epidermal growth factor receptor (EGFR). Although EGFR tyrosine kinase inhibitors (TKIs) are available clinically, the molecular complexity of NSCLC has made it necessary to search for alternative therapeutic approaches to overcome the drug resistance of NSCLC. In the present study, we identified a triterpene molecule derived from the herbal plant Tripterygium wilfordii, celastrol, as a novel autophagy inducer. We demonstrate that celastrol exhibited selective cytotoxic effect towards EGFR mutant NSCLCs. In addition, celastrol also facilitated the autophagic degradation of Hsp90 client protein including EGFR and Akt on both EGFR wild-type and mutant NSCLCs via calcium-mediated autophagy. Blockage of celastrol-induced autophagic degradation of EGFR by autophagic inhibitor or calcium chelator decreased celastrol-mediated cell death in gefitinib-resistant NSCLCs. Overall, our findings suggest that celastrol may be developed as an effective anticancer agent for treatment of gefitinib-resistant NSCLC in the future.