The mechanisms accounting for anticancer activity of AZD9291 (osimertinib or TAGRISSO), an approved third-generation EGFR inhibitor, in EGFR-mutant non-small cell lung cancer (NSCLC) cells and ...particularly for the subsequent development of acquired resistance are unclear and thus are the focus of this study.
AZD9219-resistant cell lines were established by exposing sensitive cell lines to AZD9291. Protein alterations were detected with Western blotting. Apoptosis was measured with annexin V/flow cytometry. Growth-inhibitory effects of tested drugs were evaluated
with cell number estimation and colony formation assay and
with mouse xenograft models. Protein degradation was determined by comparing protein half-lives and inhibiting proteasome. Gene knockdown were achieved with siRNA or shRNA.
AZD9291 potently induced apoptosis in EGFR-mutant NSCLC cell lines, in which ERK phosphorylation was suppressed accompanied with Bim elevation and Mcl-1 reduction likely due to enhanced Mcl-1 degradation and increased Bim stability. Blocking Bim elevation by gene knockdown or enforcing Mcl-1 expression attenuated or abolished AZD9291-induced apoptosis. Moreover, AZD9291 lost its ability to modulate Bim and Mcl-1 levels in AZD9291-resistant cell lines. The combination of a MEK inhibitor with AZD9291 restores the sensitivity of AZD9291-resistant cells including those with C797S mutation to undergo apoptosis and growth regression
and
Modulation of MEK/ERK-dependent Bim and Mcl-1 degradation critically mediates sensitivity and resistance of EGFR-mutant NSCLC cells to AZD9291 and hence is an effective strategy to overcome acquired resistance to AZD9291.
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N-alkane-based proxies are widely employed to reconstruct paleoclimate and paleoenvironment in lacustrine environments. However, little is known about the influence of microbially mediated ...alkane-degradation on n-alkane-derived proxies. In this study, the chemical composition of n-alkanes and microbially mediated n-alkane degradation potential were investigated in the surface sediment samples collected from seven lakes with a range of salinity from freshwater to salt saturation on the northern Qinghai-Tibetan Plateau (QTP). The results showed that the chemical composition of n-alkanes differed among the studied QTP lakes. Significant correlations were observed between salinity and some n-alkane-based paleoclimate and paleoenvironment proxies, such as ratio of C21−/C22+, average chain length (ACL) and carbon preference index (CPI). This suggested that salinity may affect the validity of some n-alkane-based paleoclimate and paleoenvironment proxies. Alkane-degrading bacteria were abundant and widespread in the studied freshwater and saline/hypersaline lakes but were minor or absent in salt-saturation lakes. The obtained alkane-degrading bacterial strains showed active ability to degrade n-hexadecane. This suggested that the salinity influence on the n-alkane distribution may be partially related to microbial degradation, which awaits further in-situ investigation. So salinity variation should be taken into account when using n-alkane-based proxies for reconstructing paleoclimate and paleoenvironment in lakes.
•Chemical composition of n-alkanes differed among the studied lakes of different salinity•Salinity may affect the validity of n-alkane-based paleoclimate proxies.•Alkane-degrading bacteria showed active ability to degrade n-hexadecane.•Microbial degradation may partially account for salinity influence on n-alkane distribution.•Salinity variation should be taken into account when using n-alkane-based proxies.
Bridge failure, which is generally associated with serious economic and life losses, is defined as the incapacity of a constructed bridge or its components to perform as specified in the design and ...construction requirements. This paper presents an overview of current researches on the typical characteristics and causes of bridge failures based on 10 former investigations. Principal causes can be divided into internal causes and external causes or natural factors and human factors. Design error, construction mistakes, hydraulic, collision, and overload are the top 5 leading causes of bridge failures, resulting in more than 70% of the bridge failures. Causes of bridge failures are closely related to regional economy, structural type, type of use, material type, and service age. The failure rate is very high for steel bridges, which is inseparable from excessive emphasis on structure strength but lack of consideration on structure stability and fatigue in early years. Researchers need to strengthen their research on the stability and fatigue of steel bridges, as well as inspection and maintenance. Extreme loads such as flood, collision, and overload contribute to a large number of bridge failures because of the lack of extreme loads data and design theory defects. It is critical for such bridges to have sufficient redundancy and capacity protection measures to reduce the probability of bridge failure due to extreme loads. Previous statistical methods and classification methods for the characteristics and causes of bridge failures lack unified standards, and a more scientific method needs to be established. A comprehensive electronic database on bridge damage and failures needs to be developed to establish damage models and conduct forensic studies to improve the design theory and specifications.
•Statistical characteristics and causes of bridge failure are reviewed.•Standard for classification of causes and characteristics of bridge failure is needed.•The causes of bridge failures are related to the regional economy, structural type, type of use, material type, and service age.•Design error, construction mistakes, hydraulic, collision and overload are the top 5 leading causes of bridge failures.•It is critical for bridges to have sufficient redundancy and capacity protection measures to reduce the probability of bridge failure due to extreme loads.
Loss of LKB1 is associated with increased metastasis and poor prognosis in lung cancer, but the development of targeted agents is in its infancy. Here we report that a glutaminolytic enzyme, ...glutamate dehydrogenase 1 (GDH1), upregulated upon detachment via pleomorphic adenoma gene 1 (PLAG1), provides anti-anoikis and pro-metastatic signals in LKB1-deficient lung cancer. Mechanistically, the GDH1 product α-KG activates CamKK2 by enhancing its substrate AMPK binding, which contributes to energy production that confers anoikis resistance. The effect of GDH1 on AMPK is evident in LKB1-deficient lung cancer, where AMPK activation predominantly depends on CamKK2. Targeting GDH1 with R162 attenuated tumor metastasis in patient-derived xenograft model and correlation studies in lung cancer patients further validated the clinical relevance of our finding. Our study provides insight into the molecular mechanism by which GDH1-mediated metabolic reprogramming of glutaminolysis mediates lung cancer metastasis and offers a therapeutic strategy for patients with LKB1-deficient lung cancer.
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•GDH1 contributes to anoikis resistance and tumor metastasis by activating CamKK2•α-KG binds to CamKK2 and recruits AMPK to CamKK2•In LKB1 null lung cancer, GDH1-induced CamKK2 substitutes for LKB1 to activate AMPK•PLAG1 induces GDH1 expression upon cell detachment from the matrix
Although elevated glutaminolysis has been demonstrated in cancer cells, the precise mechanism by which glutaminolysis promotes tumor metastasis remains unclear. In this article, Jin et al. demonstrate a mechanism by which GDH1 provides anti-anoikis and pro-metastatic signals through activating CamKK2 and AMPK that promotes tumor metastasis in LKB1-deficient lung cancer.
Treatment of EGFR-mutant non-small cell lung cancer (NSCLC) with mutation-selective third-generation EGFR-tyrosine kinase inhibitors (EGFR-TKIs) such as osimertinib has achieved remarkable success in ...the clinic. However, the immediate challenge is the emergence of acquired resistance, limiting the long-term remission of patients. This study suggests a novel strategy to overcome acquired resistance to osimertinib and other third-generation EGFR-TKIs through directly targeting the intrinsic apoptotic pathway. We found that osimertinib, when combined with Mcl-1 inhibition or Bax activation, synergistically decreased the survival of different osimertinib-resistant cell lines, enhanced the induction of intrinsic apoptosis, and inhibited the growth of osimertinib-resistant tumor in vivo. Interestingly, the triple-combination of osimertinib with Mcl-1 inhibition and Bax activation exhibited the most potent activity in decreasing the survival and inducing apoptosis of osimertinib-resistant cells and in suppressing the growth of osimertinib-resistant tumors. These effects were associated with increased activation of the intrinsic apoptotic pathway evidenced by augmented mitochondrial cytochrome C and Smac release. Hence, this study convincingly demonstrates a novel strategy for overcoming acquired resistance to osimertinib and other 3rd generation EGFR-TKIs by targeting activation of the intrinsic apoptotic pathway through Mcl-1 inhibition, Bax activation or both, warranting further clinical validation of this strategy.
Highlights • Met amplification and protein hyperactivation is a resistance mechanism to both 1st and 3rd generation EGFR inhibitors • Met inhibition effectively overcomes the resistance to 3rd ...generation EGFR inhibitors both in vitro and in vivo, including enhancement of apoptosis or G1 cell cycle arrest. • Enhanced Bim stabilization is a critical mechanism that mediates augmented induction of apoptosis by combination of Met inhibition with a 3rd generation EGFR inhibitor.
Anthocyanin synthesis and degradation processes were analyzed at transcript, enzyme, and metabolite levels to clarify the effects of high temperature on the concentration of anthocyanin in plum fruit ...(Prunus salicina Lindl.). The transcript levels of PsPAL, PsCHS, and PsDFR decreased while those of PsANS and PsUFGT were similar at 35 °C compared with 20 °C. The activities of the enzymes encoded by these genes were all increased in fruits at 35 °C. The concentrations of anthocyanins were higher at 35 °C on day 5 but then decreased to lower values on day 9 compared with that at 20 °C. Furthermore, high temperature (35 °C) increased the concentration of hydrogen peroxide and the activity of class III peroxidase in the fruit. The concentration of procatechuic acid, a product of the reaction between anthocyanin and hydrogen peroxide, hardly changed at 20 °C but was significantly increased at 35 °C on day 9, indicating that anthocyanin was degraded by hydrogen peroxide, which was catalyzed by class III peroxidase. Based on mathematical modeling, it was estimated that more than 60-70% was enzymatically degraded on day 9 when the temperature increased from 20 °C to 35 °C. We conclude that at the high temperature, the anthocyanin content in plum fruit depend on the counterbalance between its synthesis and degradation.
We aimed to understand the molecular mechanism underlying the incidence of Oxaliplatin resistance in colorectal cancer. The Oxaliplatin-resistant (OR) HT29 colorectal cell line was established by ...long-term exposure to Oxaliplatin. Cell viability and proliferation were determined by the 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide and direct counting assays, respectively. Transcript level of metallothionein 2A (MT2A) was measured by real-time polymerase chain reaction. Protein levels of MT2A, BRCA1-associated RING domain 1 (BARD1), BRCA1, and β-actin were quantified by immunoblotting. Direct interaction between MT2A with BARD1 and BRCA1 was analyzed by co-immunoprecipitation. Colocalization between of MT2A and BARD1 was determined by immunofluorescence. MT2A was upregulated in OR cells at both transcript and protein levels. Knockdown of MT2A in HT29 OR cells improved sensitivity to Oxaliplatin, while ectopic overexpression of MT2A conferred HT29 cells relative resistance to Oxaliplatin. We further demonstrated that MT2A interacted with and positively regulated BARD1/BRCA1 in colorectal cancer cells. BARD1 overexpression partially restored the compromised Oxaliplatin resistance elicited by MT2A deficiency in terms of both cell proliferation and viability. Our data highlighted the critical contributions of MT2A-BARD1/BRCA1 in Oxaliplatin resistance in colorectal cancer cells.
Magnetotactic bacteria (MTB) can rapidly relocate to optimal habitats by magnetotaxis, and play an important role in iron biogeochemical cycling. This study aimed to evaluate the contribution of the ...external magnetostatic field to the diversity of MTB in freshwater sediments from Yangtze River (Changjiang River, CJ), Chagan Lake (CGH) and Zhalong Wetland (ZL). The magnetic field intensity was tightly associated with the community richness of MTB in CJ, whereas it was closely related to the diversity of MTB in CGH and ZL (p < 0.05), elucidating a significant variation in the community composition of MTB. Magnetic exposure time appeared more significant correlation with community richness than diversity for MTB in CJ and CGH (p < 0.05), while an opposite relationship existed in ZL (p < 0.01). Herbaspirillum (93.81–96.48 %) dominated in the sediments of these surfacewatesr regardless of waterbody types, while it shifted to Magnetospirillum in ZL under 100 Gs magnetic field. The network connectivity and stability of MTB deteriorate with the increase of magnetic field intensity. Functional analysis showed that the Two-component system and ABC transporter system of MTB obviously responded to magnetic field intensity and exposure time. Our findings will pave the way to understanding the response mechanism of MTB community in freshwater sediments to the external magnetostatic field.
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•MTB widely exist in Changjiang River (CJ), Chagan Lake (CGH) and Zhalong Wetland (ZL) of China.•Dominant Herbaspirillum and Magnetospira occur all sediment samples.•Magnetic field intensity and exposure time both affect MTB diversity and richness.•Network connectivity and stability of MTB deteriorate as magnetic field intensity.•Metabolic function and ABC transporter system visibly respond to magnetic field.
Little is known about the relative importance of spatial and environmental factors to structuring aquatic and sedimentary microbial biogeography in lakes. Here, we investigated the microbial ...community composition (MCC) of the water (
= 35) and sediment (
= 35) samples from 16 lakes in western China (salinity: freshwater to salt saturation; pairwise geographical distance: 9-2027 km) using high-throughput sequencing and evaluated the relative importance of spatial and environmental factors to microbial (including total, abundant, and rare) distributions. Our results showed that spatial factors were more important than environmental factors in shaping the biogeography of aquatic and sedimentary microbial communities in the studied lakes, and spatial factors on abundant microbial community was stronger than that on the total/rare microbial communities. Moreover, sedimentary rare MCC might be more sensitive to environmental factors than its aquatic counterpart. Such different biogeography responses of total, abundant, and rare communities to environmental and spatial factors could be ascribed to different physiochemical properties between water and sediment. Collectively, this study expands our understanding of factors shaping microbial biogeography of total, abundant, and rare communities between waters and sediments of lakes.
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