Pancreatic cancer (PC) is one of the most lethal cancers known worldwide, and its prognosis is poor in most patients. Exosomes are nanosized extracellular vesicles, which are released from various ...cell types. They are involved in cellular communication. The diagnosis and treatment of PC were improved substantially with exosomes. In this study, we isolated PC‐derived exosomes and investigated their proteomic profile. Then, we conducted bioinformatic analysis on proteomic data. Differential ultracentrifugation was performed to isolate exosomes from human serum samples and four PC cell lines. Transmission electron microscopy and Western blot analysis were used to characterize the isolated exosomes. Liquid chromatography coupled with tandem mass spectrometry was conducted to identify the proteome of serum exosomes. Proteomic analysis demonstrated that all the serum exosomes were derived from three cohorts of human subjects; these serum exosomes contained a total of 655 proteins, out of which 315 proteins overlapped with ExoCarta database. Gene oncology and kyoto encyclopedia of genes and genomes analyses provided the functional annotation of the proteome. Interestingly, 18 or 14 proteins were upregulated and 11 or 14 proteins were downregulated in serum exosomes derived from patients with PC as compared with in serum exosomes derived from healthy volunteers or from pancreatitis patients respectively. Annexin A11, a calcium‐dependent phospholipid‐binding protein, was expressed in a PC cell line (CFPAC‐1)‐derived exosomes and in tumor tissues of patients with PC, respectively. Our data provided a basic foundation for further studies on the protein composition of PC‐derived exosomes and its involvement in PC biology.
The emergence of a vast array of visual artworks meets the growing public demand for cultural literacy while simultaneously presenting challenges in the understanding and selection of appropriate ...artistic styles. Leveraging advancements in image processing technology, this study successfully automates the classification of 158,652 visual art images by categorizing them into distinct artistic styles. A novel approach combining the Scale-Invariant Feature Transform (SIFT) feature extraction algorithm with the spatial pyramid matching technique is introduced. This paper details the development and evaluation of a classification system tailored for the automatic categorization of visual artworks. Comparative testing results demonstrate that this innovative combination surpasses the traditional SIFT algorithm in both execution speed and classification accuracy. Specifically, the Support Vector Machine (SVM) based classification system, utilizing the enhanced SIFT method, achieves an accuracy rate exceeding 90% in identifying five distinct styles, including PHPS, NOS, IS, TF, and AAS. The implementation of this automatic classification system proves to be a practical tool in aiding individuals to select the visual artwork style that best suits their preferences.
Cannabis research has historically focused on the most prevalent cannabinoids. However, extracts with a broad spectrum of secondary metabolites may have increased efficacy and decreased adverse ...effects compared to cannabinoids in isolation. Cannabis's complexity contributes to the length and breadth of its historical usage, including the individual application of the leaves, stem barks, and roots, for which modern research has not fully developed its therapeutic potential. This study is the first attempt to profile secondary metabolites groups in individual plant parts comprehensively. We profiled 14 cannabinoids, 47 terpenoids (29 monoterpenoids, 15 sesquiterpenoids, and 3 triterpenoids), 3 sterols, and 7 flavonoids in cannabis flowers, leaves, stem barks, and roots in three chemovars available. Cannabis inflorescence was characterized by cannabinoids (15.77-20.37%), terpenoids (1.28-2.14%), and flavonoids (0.07-0.14%); the leaf by cannabinoids (1.10-2.10%), terpenoids (0.13-0.28%), and flavonoids (0.34-0.44%); stem barks by sterols (0.07-0.08%) and triterpenoids (0.05-0.15%); roots by sterols (0.06-0.09%) and triterpenoids (0.13-0.24%). This comprehensive profile of bioactive compounds can form a baseline of reference values useful for research and clinical studies to understand the "entourage effect" of cannabis as a whole, and also to rediscover therapeutic potential for each part of cannabis from their traditional use by applying modern scientific methodologies.
Previous phylogenetic analyses of species within the genus Golovinomyces (Ascomycota, Erysiphales), based on ITS and 28S rDNA sequence data, revealed a co-evolutionary relationship between powdery ...mildew species and hosts of certain tribes of the plant family Asteraceae. Golovinomyces growing on host plants belonging to the Heliantheae formed a single lineage, comprised of a morphologically differentiated complex of species, which included G. ambrosiae, G. circumfusus, and G. spadiceus. However, the lineage also encompassed sequences retrieved from Golovinomyces specimens on other Asteraceae tribes as well as other plant families, suggesting the involvement of a plurivorous species. A multilocus phylogenetic examination of this complex, using ITS, 28S, IGS (intergenic spacer), TUB2 (beta-tubulin), and CHS1 (chitin synthase I) sequence data was carried out to clarify the discrepancies between ITS and 28S rDNA sequence data and morphological differences. Furthermore, the circumscription of species and their host ranges were emended.
The phylogenetic and morphological analyses conducted in this study revealed three distinct species named, viz., (1) G. ambrosiae emend. (including G. spadiceus), a plurivorous species that occurs on a multitude of hosts including, Ambrosia spp., multiple species of the Heliantheae and plant species of other tribes of Asteraceae including the Asian species of Eupatorium; (2) G. latisporus comb. nov. (≡ Oidium latisporum), the closely related, but morphologically distinct species confined to hosts of the Heliantheae genera Helianthus, Zinnia, and most likely Rudbeckia; and (3) G. circumfusus confined to Eupatorium cannabinum in Europe.
The present results provide strong evidence that the combination of multi-locus phylogeny and morphological analysis is an effective way to identify species in the genus Golovinomyces.
Soil salinization seriously restricts the growth and yield of soybeans. However, little information is available on the early growth stages of soybeans which are subjected to the gibberellin ...biosynthesis inhibitor, prohexadione-calcium (Pro-Ca). This study aimed to investigate the effects of exogenous Pro-Ca on saline-alkali stress-induced damages to photosynthesis and antioxidant defenses in soybean (Glycine max L.) seedlings. At the V3 growth stage, salt-tolerant genotype Hefeng 50 (HF50) and salt-sensitive genotype Kenfeng 16 (KF16) were subjected to 110 mmol L−1 mixed saline-alkali stress respectively, and then 100 mg L−1 Pro-Ca was sprayed on the leaves. Our results showed that saline-alkali stress accelerated the degradation of thylakoids, inhibited chlorophyll synthesis, reduced shoot dry weight, electron transfer rate (ETR), and peroxidase (POD) activity, the concentration of ascorbic acid (AsA) and soluble sugar, but enhanced the concentration of proline, hydrogen peroxide (H2O2) and the rate of superoxide radical (O2∙−) generation. Additionally, saline-alkali stress induced a lower decrease of the net photosynthetic rate (Pn), potential activity of PSII (Fv/F0), and maximum quantum yield of PSII (Fv/Fm) in salt-tolerant HF50 than in salt-sensitive KF16. Nevertheless, foliar spraying of exogenous Pro-Ca increased the chlorophyll content, Pn, Fv/F0, and Fv/Fm. These results were more prominent when Pro-Ca was applied to KF16 under saline-alkali conditions. Furthermore, exogenous application of Pro-Ca retarded the degradation of thylakoids, increased the ETR and the accumulation of AsA, soluble sugar, and proline, activated the activities of superoxide dismutase (SOD), catalase (CAT), and POD, and decreased the concentration of malondialdehyde (MDA), electrolyte leakage (EL), O2∙−, and H2O2. These results indicated that Pro-Ca could effectively protect soybean seedlings against damage from saline-alkali stress by regulating seedling phenotype, photosynthetic apparatus, antioxidant defense, and osmoregulation.
•Application of prohexadione-calcium alleviated the saline-alkali stress in soybean seedlings.•Prohexadione-calcium application inhibited the degradation of chlorophyll and thylakoid.•Prohexadione-calcium application upregulated the activities of antioxidant enzymes and the level of non-antioxidants.•Prohexadione-calcium application promoted the accumulation of soluble sugar and proline in soybean leaf.
Highly upregulated in liver cancer (HULC), a lncRNA that is considered a key molecule in human liver cancer, has recently been revealed to be involved in hepatocellular carcinoma (HCC) development ...and progression 1, 2. It has been reported that HULC can promote tumor invasion and metastasis of HCC, but its function and mechanism of action in HCC have not been elucidated. In this study, we found that HULC was aberrantly up-regulated in HCC tissues and associated with TNM stage, intrahepatic metastases, HCC recurrence, and postoperative survival. HULC depletion inhibited the growth and metastasis of HCC cell lines in vitro and in vivo. Moreover, HULC contributes to ZEB1-induced epithelial-mesenchymal transition (EMT), a requirement for tumor invasion and metastasis that plays a key role in cancer progression. This effect of ZEB1 was inhibited by HULC siRNA. We conclude that the HULC functioned as a competing endogenous RNA (ceRNA) to mediate EMT via up-regulating ZEB1. In this way, it sequesters the miR-200a-3p signaling pathway to facilitate HCC metastasis. HULC comes into play as an oncogene in HCC, acting mechanistically by inducing HCC cells to activate EMT. Such an effect promotes tumor progression and metastasis through the miR-200a-3p/ZEB1 signaling pathway. The identification of this novel pathway that links high expression levels of HULC with EMT in HCC cells may serve as the foundation for the development of novel anti-tumor therapeutics.
Abstract
Background
The importance of mRNA methylation erased by ALKBH5 in mRNA biogenesis, decay, and translation control is an emerging research focus. Ectopically activated YAP is associated with ...the development of many human cancers. However, the mechanism whereby ALKBH5 regulates YAP expression and activity to inhibit NSCLC tumor growth and metastasis is not clear.
Methods
Protein and transcript interactions were analyzed in normal lung cell and NSCLC cells. Gene expression was evaluated by qPCR and reporter assays. Protein levels were determined by immunochemical approaches. Nucleic acid interactions and status were analyzed by immunoprecipitation. Cell behavior was analyzed by standard biochemical tests. The m
6
A modification was analyzed by MeRIP.
Results
Our results show that YAP expression is negatively correlated with ALKBH5 expression and plays an opposite role in the regulation of cellular proliferation, invasion, migration, and EMT of NSCLC cells. ALKBH5 reduced m
6
A modification of
YAP
. YTHDF3 combined
YAP
pre-mRNA depending on m
6
A modification. YTHDF1 and YTHDF2 competitively interacted with YTHDF3 in an m
6
A-independent manner to regulate
YAP
expression. YTHDF2 facilitated
YAP
mRNA decay via the AGO2 system, whereas YTHDF1 promoted
YAP
mRNA translation by interacting with eIF3a; both these activities are regulated by m
6
A modification. Furthermore, ALKBH5 decreased YAP activity by regulating miR-107/LATS2 axis in an HuR-dependent manner. Further, ALKBH5 inhibited tumor growth and metastasis in vivo by reducing the expression and activity of YAP.
Conclusions
The presented findings suggest m
6
A demethylase ALKBH5 inhibits tumor growth and metastasis by reducing YTHDFs-mediated YAP expression and inhibiting miR-107/LATS2–mediated YAP activity in NSCLC. Moreover, effective inhibition of m
6
A modification of ALKBH5 might constitute a potential treatment strategy for lung cancer.
MYH9 encodes the heavy chain of nonmuscle myosin IIA, a ubiquitously expressed cytoplasmic myosin that regulates the actin cytoskeleton, cell migration, cell polarization, and signal transduction in ...cancer cells. Here, we investigated the role of MYH9 in cancer stem cells (CSCs) associated with esophageal cancer (EC). The subcellular localization of MYH9 was investigated in SKGT‐4 cells through immunofluorescent analysis. MYH9+ and MYH9− spheroid cells were derived from SKGT‐4 cells by flow cytometry and compared for self‐renewal capacity, tumorigenicity, CD133 positivity, cisplatin resistance, and phosphatidylinositol‐3‐kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) activity. MYH9 messenger RNA expression was assessed in 30 EC patients by quantitative reverse transcription‐polymerase chain reaction. Kaplan–Meier curves were plotted to explore the influence of MYH9 on EC survival. MYH9 localized to the plasma membrane, cytoplasm, and nucleus of SKGT‐4 cells. Spheroid cells displayed higher MYH9 expression and positivity compared to parental SKGT‐4 cells. MYH9+ cells showed strong CSC characteristics, including in vivo tumorigenicity, migration, invasion, cisplatin resistance, and CD133+ positivity. MYH9 activated the PI3K/AKT/mTOR axis in CSCs and was upregulated in EC patients with poor survival. Collectively, these data show that MYH9 significantly promotes tumorigenesis by regulating PI3K/AKT/mTOR signaling in EC. MYH9 expression remarkably correlates with poor prognosis and represents a novel biomarker and drug target for the diagnosis and treatment of EC.