P388 Cells Research Articles

Secondary metabolites from Colletotrichum gloeosporioides isolated from Artocarpus heterophyllus and evaluation of their cytotoxic and antibacterial activities

A new lactone, collectindolide (1), as well as three known compounds, (S)-sydonic acid (2), (S)-hidroxysydonic acid (3), and indole-3-aceticacid (4) were obtained from the liquid media of Colletotrichum gloeosporioides, which was obtained from Artocarpus heterophyllus. The chemical structures of 1-4 were established by spectroscopic analyses, including NMR experiments and by HR-ESI-TOF-MS mass spectroscopy. Compounds 1-4 were evaluated for their cytotoxicity against murine leukaemia P-388 cell lines by MTT assay. Antibacterial activity of compounds 1-4 was also assayed against four bacteria. Phytochemical investigation of the genus Colletotrichum derived from the plant genus Artocarpus is reported for the first time.

PLCL1 suppresses tumour progression by regulating AMPK/mTOR-mediated autophagy in renal cell carcinoma.

Autophagy has been increasingly recognized as a critical regulatory mechanism in the maintenance of cellular homeostasis. A previous study showed that phospholipase C-like protein 1 (PLCL1) is associated with lipid metabolism in renal cell carcinoma (RCC). However, it is unclear whether PLCL1 regulates autophagy, thereby influencing the progression of RCC. Bioinformatics analysis of five microarray datasets revealed that expression of PLCL1 is decreased in tumours and is positively correlated with prognosis in RCC patients. Three independent public datasets, clinical RCC tissues and RCC cell lines, were validated using real-time qPCR, western blotting and immunohistochemistry. Using wound healing and transwell assays, we observed that elevated PLCL1 levels decreased the migratory distance and the invasive number of 786-O and ACHN cells, but PLCL1 knockdown reversed these changes in 769P cell lines compared to those in controls. The results of flow cytometry analysis indicated that PLCL1 promotes apoptosis. Moreover, transcriptional analysis based on stable overexpression of PLCL1 in 786-O cells revealed that PLCL1 is related to autophagy, and western blotting and autophagic experimental results further verified these findings. Mechanistic investigations confirmed that PLCL1 activates the AMPK/mTOR pathway and interacts with decidual protein induced by progesterone (DEPP). Collectively, our data suggest that PLCL1 functions as a suppressor of RCC progression by activating the AMPK/mTOR pathway, interacting with DEPP, initiating autophagy and inducing apoptosis. PLCL1 may be a promising therapeutic target for the diagnosis and treatment of ccRCC patients.

Diterpenoid WT-29 isolated from Wedelia exerted anti-inflammatory and anti-allergic activities

Ethnopharmacological relevanceWedelia (Sphagneticola trilobata) is a traditional anti-inflammatory herb native to tropical America. It is commonly used to treat some inflammatory related diseases clinically, such as pertussis, pharyngitis, etc. However, its specific anti-inflammatory mechanism is still unclear. Aim of the studyWT-29 (3α-angeloyloxy-9β-hydroxyent-kaura-16-en-19-oic acid) is a main bioactive diterpenoid isolated and purified from Wedelia. This study aims to explore the potential anti-inflammatory and anti-allergic properties of WT-29 on RAW264.7 cells stimulated with LPS and P815 cells induced by C48/80, as well as investigating their underlying molecular mechanisms. MethodsThe anti-inflammatory mechanism of WT-29 was analyzed and predicted using network pharmacology, and then verified through experiments. The Griess reagent assay was employed to evaluate the impact of WT-29 on the generation of nitric oxide (NO) in RAW264.7 cells induced by LPS, the expression of various inflammatory cytokines and the release of histamine in cells were measured through qRT-PCR and ELISA techniques. The impact of WT-29 on the translocation of the NF-κB p65 protein to the nucleus was assessed through immunofluorescence staining. Western blot technique was utilized to investigate protein expression in inflammation, allergy, and autophagy pathways. ResultsThe study found that WT-29 can reduce the secretion of inflammatory factors (NO, iNOS, COX-2, IL-6, IL-1β and TNF-α), inhibit NF-κB activation and MAPK family phosphorylation, and induce autophagy in RAW264.7 cells stimulated with LPS. In addition, it demonstrated that WT-29 could inhibit histamine release and degranulation, as well as inhibit the MAPK family in C48/80-induced P815 cells. ConclusionWT-29 isolated from Wedelia exerts anti-inflammatory and anti-allergic effects mainly through NF-κB, Nrf2/Keap-1, MAPK pathways and regulating of autophagy, suggesting that it might be a potential anti-inflammatory and anti-allergic agent and could be used as medicine or health benefit product.

Botryorhodines K and L, two new cytotoxic depsidones from a fungus of the genus Arcopilus.

Botryorhodines K (1) and L (2), two new depsidone derivatives, along with one known metabolite, 4-O-demethylbarbatic acid (3), were isolated from the culture extract of a fungus of the genus Arcopilus. The structures of 1‒3 were determined by the analysis of NMR and MS spectral data and the absolute configuration of 1 was established by single-crystal X-ray diffraction analysis. Compounds 1 and 2 showed antimicrobial activity against Gram-positive bacteria and cytotoxicity against murine leukemia P388 cells.

Discovery of Four New Compounds from Macropanax membranifolius and Their Cytotoxic Activity.

A phytochemical investigation of the methanolic extract of the Macropanax membranifolius C.B. Shang leaves led to the isolation of three new flavans, (2R,3R)-4'-O-methylcatechin 5-O-β-D-glucopyranoside (1), (2S,3S)-4'-O-methylcatechin 5-O-β-D-glucopyranoside (2), (2S,3R)-4'-O-methylcatechin 5-O-β-D-glucopyranoside (3), one new triterpene glycoside 3-O-β-D-xylopyranosyl-(1→6)-[β-D-xylopyranosyl-(1→2)]-β-D-glucopyranosyl-oleanolic acid 28-O-β-D-glucopyranoside (4), together with nine known compounds (5-13). Their chemical structures were elucidated based on HR-ESI-MS, NMR spectroscopic data. The absolute configurations of compounds 1-3 were established by electronic circular dichroism (ECD) spectra. At concentration of 20 μM, compounds 1-13 showed the percentages of dead cell in the range of 2.14 % to 33.61 % against KB, HepG2, HL60, P388, HT29, and MCF7 cancerous cell lines by SRB assay.

SEMA3G functions as a novel prognostic biomarker associated with Wnt pathway in clear cell renal cell carcinoma

Renal cell cancer (RCC) is one of the most common cancer, and the incidence of clear cell renal cell cancer rank at the first among multiple subtypes of RCC. Tumor heterogeneity and limited therapies expedite researches and studies on prognostic biomarkers and molecular mechanism. SEMA3G mediates various bimolecular processes but few studies have assessed the influence of SEMA3G on ccRCC. The expression of SEMA3G at mRNA level in ccRCC was analyzed using 4 TCGA datasets. The expression at protein level was verified by immunohistochemistry and western blot. Biological pathway was explored by GSEA and western blot. At both mRNA and protein level, SEMA3G expressed significantly lower in ccRCC tissues compared with normal renal tissues, and the expression was highly associated with clinical stage and pathological grade. Low expression of SEMA3G indicated a poorer overall survival and disease specific survival. Transwell and wound-healing assays showed that overexpressed SEMA3G inhibited the cell motility of renal cancer cells. Upregulated SEMA3G suppressed the invasion and proliferation of both 769-P and 786-O cells. Wnt signaling pathway was tested to work in the interfering of SEMA3G on tumorigenesis and progression of ccRCC. The results provide novel insight into the role of SEMA3G in ccRCC, suggesting the prognostic value and potential suppressor role of SEMA3G.

New indole alkaloid Morucolletotricin from endophytic fungus Colletotrichum queenslandicum associated with Morus australis Poir. Leaf

Six compounds were isolated from the ethyl acetate extract of endophytic fungus Colletotrichum queenslandicum derived from Morus australis Poir. leaf. Based on NMR and MS data led to characterised of these compounds including one new indole alkaloid Morucolletotricin (1) along with two other indole alkaloids; tryptopol (2) and indole-3-acetic acid (3), phomopyronol (4), 2-(3-aminophenyl)acetic acid (5) and ergosterol (6). phomopyronol (4) and 2-(3-aminophenyl)acetic acid (5) were first reported from Colletotrichum fungi. The cytotoxic activity of compounds 1-6 was evaluated against murine leukaemia P-388 cells and showed that all compounds were moderate cytotoxic. phomopyronol (4) was the most active among the five other compounds with IC50 = 37.17 μg/mL.

Comprehensive analysis of E47‑like factors and verification of ELF4 in clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) is the most prominent subtype of renal cancer and E47-like factors (ELFs) are important in tumorigenesis; however, the specific role of key ELFs in ccRCC remains unclear. The present study comprehensively analyzed RNA sequencing and clinical data from multiple databases, and identified differentially expressed ELFs (ELF3-5) in ccRCC. The DNA promoter methylation, genetic variation and clinical significance of ELF3-5 in ccRCC were analyzed using the cBioPortal and UALCAN databases. The association between ELF3-5 and multiple immune cell infiltration was analyzed using Tumor Immune Estimation Resource. Subsequently, ELF4 was selected and its association with biological functions was assessed. Cell counting kit-8 (CCK-8), colony formation, Transwell, macrophage chemotaxis and polarization assays were conducted to validate the functions of ELF4. Notably, the mRNA expression levels of ELF4 were significantly upregulated in ccRCC, whereas ELF3 and ELF5 mRNA expression levels were significantly downregulated. Clinical significance analysis revealed that ELF4 showed a high clinical significance with tumor grade, clear cell type A and B subtypes, and incidence rates of amplification in genetic variation. Further analyses indicated that ELF4 may be involved in multiple immune cell differentiation. Additionally, cell experiments revealed that ELF4 inhibition downregulated 769-P and 786-O proliferation, migration and invasion. Knockdown of ELF4 in cancer cells also inhibited M2 macrophage polarization and chemotaxis towards 769-P and 786-O cells. Conclusively, the present findings indicated the clinical significance of ELF4 in ccRCC, and verified its key role in driving cell proliferation, migration and invasion, and promoting M2 macrophage polarization and chemotaxis in ccRCC.

A Novel Tumor Mutation Burden Related lncRNA Signature Identified Prognosis and Tumor Immune Microenvironment Features in Clear Cell Renal Cell Carcinoma.

Emerging evidence indicates that long noncoding RNA (lncRNA) plays an important biological role in clear cell renal cell carcinoma (ccRCC); however, the clinical value of tumor mutation burden-related lncRNA in ccRCC patients is unknown yet. Somatic mutation profiles and lncRNA expression data of ccRCC were downloaded from the TCGA database. We retrospectively analyzed lncRNA expression data and survival information from 116 patients with ccRCC fromJanuary 2013 to January 2014. Univariate and multivariate Cox regression analyses were performed to construct lncRNA signature, and the prognosis value was determined by Kaplan-Mayer and receiver operating characteristic curve (ROC) analysis. Based on 160 differentially expressed TMB-related lncRNAs, two TMB-related molecular clusters with distinct immune checkpoints expression and immune cells infiltration were established for ccRCC patients. Moreover, a novel TMB-related lncRNA signature was constructed based on five lncRNAs for individualized prognosis assessment. High-risk group represents significantly worse overall survival in all cohorts. The area under the ROC curve was 0.716, 0.775 and 0.744 in the training cohort, testing cohort and TCGA cohort, respectively. Results of qRT-PCR successfully validated the expression levels of AP002360.3, LINC00460, AL590094.1, LINC00944 and LINC01843 in HK-2, 786-O, 769-P and ACHN cells. More importantly, the predictive performance of TMB-related lncRNA signature was successfully validated in an independent cohort of 116 ccRCC patients at our institution. This study successfully developed and validated a novel TMB-related lncRNA signature for individualized prognosis assessment of ccRCC patients.

Development of A Plant-Based Meat Product Supplemented With Red Amaranth Antioxidants For The Elderly

This study was aimed at developing a protein-rich food formulation for the elderly using ingredients derived from soybean, sacha inchi, wheat flour, quinoa, and perilla seed. First, the protein content of all ingredients was analyzed. The results showed that the highest protein level (48.54%) was seen in sacha inchi. Then, sensory test by elderly adults was evaluated. The formulation which had the highest sensory acceptance comprised 33% soybean, 40% sacha inchi, 20% wheat flour, 5% quinoa, and 2% perilla seed. The effect of stabilizers (xanthan gum and sodium alginate) at levels of 0.1 and 0.2%, respectively, was studied. It was found that 0.1% sodium alginate produced the highest sensory score. Measurements of the texture and water absorption of the formulation showed that the values for hardness, cohesiveness, springiness, adhesiveness, chewiness and gumminess were 1003, 0.25, 0.45, 0.17, 110, and 222, respectively, while water absorption was 51.10%. An aqueous extract of Amaranthus dubius was subjected to analysis of levels of polyphenols and anthocyanins, as well as antioxidant capacity and cytotoxicity. The aqueous extract had polyphenol, anthocyanin, DPPH radical-scavenging and FRAP levels of 41.13 µg GAE/mL, 458 mg/L, 62.7%, and 14.8 µg Trolox/mL, respectively. At a concentration of 2000 µg/mL, the crude extract exerted 22% average anti-proliferative effect on P388, KB, Hela and HepG2 cells. Different extract levels were added to the product, and the acceptability of the concentrated extract was re-evaluated. The results showed that the concentrated extract at 0.5% level of incorporation had the highest acceptance rating as a meat analogue. The energy per 100 g of the plant meat sample was 247.95 kcal, while its contents of protein, fat, carbohydrate, ash, and dietary fibre were 24.71, 6.27, 23.17, 2.52 and 6.76%, respectively. Thus, the plant meat product supplemented with A. dubius extract could be an alternative and safe diet for the elderly.

Coumarin Derivative from the Unripe Fruit of Mengkudu (Morinda citrifolia Linn) and Cytotoxic Activity

A coumarin derivative namely scopoletin (7-hydroxy-6-methoxychromen-2-one)had been isolated from the methanol extract of unripe noni fruits (Morinda citrifolia Linn.). The extraction was conducted by maceration, continuing with separation and purification using several techniques of chromatography. The yellowish isolated compound showed a melting point at 200 – 202 C. The compound structure was analyzed based on spectral data 1H-NMR, 13C-NMR, and NOE 1D. The inhibition of methanol extract of noni fruits and scopoletin against murine leukemia P388 cells using MTT assay showed IC50 were > 100 μg/mL and 65.69 μg/mL respectively. These showed that scopoletin was more cytotoxic than methanol extract but weaker than artonin E as a positive control IC50 0.48 μg/mL).

Characterization of the Molecular Diversity and Degranulation Activity of Mastoparan Family Peptides from Wasp Venoms.

Wasp stings have become an increasingly serious public health problem because of their high incidence and mortality rates in various countries and regions. Mastoparan family peptides are the most abundant natural peptides in hornet venoms and solitary wasp venom. However, there is a lack of systematic and comprehensive studies on mastoparan family peptides from wasp venoms. In our study, for the first time, we evaluated the molecular diversity of 55 wasp mastoparan family peptides from wasp venoms and divided them into four major subfamilies. Then, we established a wasp peptide library containing all 55 known mastoparan family peptides by chemical synthesis and C-terminal amidation modification, and we systematically evaluated their degranulation activities in two mast cell lines, namely the RBL-2H3 and P815 cell lines. The results showed that among the 55 mastoparans, 35 mastoparans could significantly induce mast cell degranulation, 7 mastoparans had modest mast cell degranulation activity, and 13 mastoparans had little mast cell degranulation activity, suggesting functional variation in mastoparan family peptides from wasp venoms. Structure-function relationship studies found that the composition of amino acids in the hydrophobic face and amidation in the C-terminal region are critical for the degranulation activity of mastoparan family peptides from wasp venoms. Our research will lay a theoretical foundation for studying the mechanism underlying the degranulation activity of wasp mastoparans and provide new evidence to support the molecular design and molecular optimization of natural mastoparan peptides from wasp venoms in the future.

Abstract 5016: IL-6 influences PIM1 expression in renal cell carcinoma

Abstract Renal cell carcinoma (RCC) is the most common kidney neoplasm, accounting for over 430,000 new cases worldwide and 179,000 deaths annually. The overexpression of the proviral integration site for moloney murine leukemia virus 1 (PIM1) kinase is associated with poor clinical outcomes in patients with RCC. PIM1 is a constitutively active effector serine/threonine kinase with roles in tumor progression including cell proliferation, apoptosis, invasion, and migration. Yet, the mechanisms underlying PIM1 expression and its function in RCC are not fully delineated. IL-6 is a pleiotropic cytokine involved in the activation of the JAK/STAT signaling cascade. High serum IL-6 levels are implicated in the poor prognosis of RCC patients and are hypothesized to contribute to RCC invasion and metastasis. PIM1 is shown to be transcriptionally regulated downstream of JAK/STAT signaling and an IL-6/STAT3/PIM1 axis exists in pancreatic cancer. We thus hypothesize that this pathway also regulates expression of PIM1 in RCC. Here, we explore the levels of PIM1 in RCC and identify potential signaling pathways influencing PIM1 expression. Our results show five RCC cell lines, 769-P, 786-O, ACHN, CAKI-1, and RCC-4 cells overexpress PIM1 relative to normal renal proximal tubule epithelial cells. In RCC cell lines, IL-6 secretion correlates with PIM1 protein levels. Treatment with ruxolitinib, a JAK1/2 inhibitor, leads to a dose and concentration dependent decrease in PIM1 levels. We then tested whether incubation with an IL-6 neutralizing antibody is sufficient to regulate PIM1 protein levels. Indeed, use of an IL-6 antibody resulted in decreased pJAK2 protein and PIM1 protein. Our initial studies suggest that differential expression of PIM1 among RCC cell lines may be linked to autocrine IL-6 signaling. Interestingly, 769-P cells demonstrate increased PIM1 levels but do not secrete IL-6 and neither ruxolitinib treatment nor incubation with an IL-6 neutralizing antibody appear to affect PIM1 levels. Therefore, an IL-6/JAK-independent pathway resulting in increased PIM1 protein levels likely exists in 769-P cells. Further investigation is required to elucidate the contexts of PIM1 regulatory pathways in order to develop novel targeted anti-cancer strategies in RCC. Citation Format: Kimberly S. Meza, Sheldon L. Holder. IL-6 influences PIM1 expression in renal cell carcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5016.

MP04-12 PHARMACOLOGICAL INHIBITION OF THE MOLECULAR CHAPERONE HSP70 OVERCOMES BELZUTIFAN RESISTANCE IN CCRCC

You have accessJournal of UrologyCME1 Apr 2023MP04-12 PHARMACOLOGICAL INHIBITION OF THE MOLECULAR CHAPERONE HSP70 OVERCOMES BELZUTIFAN RESISTANCE IN CCRCC Michael Basin, Sarah Backe, Michael Bratslavsky, Mark Woodford, Dimitra Bourboulia, Mehdi Mollapour, and Gennady Bratslavsky Michael BasinMichael Basin More articles by this author , Sarah BackeSarah Backe More articles by this author , Michael BratslavskyMichael Bratslavsky More articles by this author , Mark WoodfordMark Woodford More articles by this author , Dimitra BourbouliaDimitra Bourboulia More articles by this author , Mehdi MollapourMehdi Mollapour More articles by this author , and Gennady BratslavskyGennady Bratslavsky More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003215.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Patients with von-Hippel Lindau (VHL) mutations are predisposed to developing clear cell renal cell carcinoma (ccRCC) through constitutive activation of transcription factor hypoxia-inducible factor 2a (HIF2a). The HIF2a antagonist Belzutifan has recently been FDA approved for treatment VHL disease associated tumors. Results of the recent clinical trial (MK-4682) revealed that a certain proportion of VHL patients progress on therapy. Our objective here was to decipher the molecular mechanisms of Belzutifan mediated downregulation of HIF2a and therefore provide better understanding of drug resistance. The molecular chaperone Hsp70 promotes the stability of hundreds of client proteins, thereby protecting them from degradation. Since HIF2a interacts with Hsp70, we hypothesized that Hsp70 maintains HIF2a stability in ccRCC and is a potential alternative therapeutic target in cases of Belzutifan resistance. METHODS: Cell line models of VHL-null ccRCC (786-0, A498, 769-P) were treated with 2 mM Belzutifan. HIF2a expression levels were evaluated by immunoblotting. Wild-type (WT) or Belzutifan resistant (S304M, G323E) HIF2a were transiently expressed in 786-O cells prior to treatment with Belzutifan or an Hsp70 inhibitor (JG-98). HIF2a degradation was examined by immunoblot. RESULTS: Here, we show that HIF2a is an Hsp70 client in VHL null ccRCC cell line 786-O. Further, we demonstrated a time dependent decrease in HIF2a expression with Belzutifan (2 mM) as early as 2 hours after treatment in three VHL null ccRCC cell lines 786-0, A498, and 769-P cells. Pretreatment of ccRCC cells with the proteasome inhibitor bortezomib blocked Belzutifan mediated downregulation of HIF2a, indicating Belzutifan leads to proteasomal degradation of HIF2a through a VHL independent pathway. Utilizing previously characterized Belzutifan resistant HIF2a-S304M and -G323E mutants we demonstrated that treating ccRCC cells with Hsp70 inhibitor JG-98 leads to mutant-HIF2a degradation, therefore overcoming Belzutifan resistance. CONCLUSIONS: The FDA-approved HIF2a inhibitor Belzutifan causes proteasomal degradation of WT-HIF2a, but not of resistant mutants. Our data indicates that HIF2a stability is reliant on the molecular chaperone Hsp70. Further, treatment with an Hsp70 inhibitor could potentially be used in the clinic to overcome resistance to Belzutifan in patients with VHL disease and ccRCC. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e37 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Michael Basin More articles by this author Sarah Backe More articles by this author Michael Bratslavsky More articles by this author Mark Woodford More articles by this author Dimitra Bourboulia More articles by this author Mehdi Mollapour More articles by this author Gennady Bratslavsky More articles by this author Expand All Advertisement PDF downloadLoading ...

Chryroxosides A–E: five new triterpene saponins from the leaves of Chrysophyllum roxburghii G.Don. and their cytotoxic activity

Five undescribed oleanane triterpene glycosides named chryroxosides A–D (1–5), together with five known compounds (6–10) were isolated from the leaves of Chrysophyllum roxburghii G.Don. Their chemical structures were elucidated by extensive spectroscopic data analyses including IR, HR-ESI-MS, 1D and 2D NMR). Compounds 1, 3, and 5 showed cytotoxic effects against KB, HepG2, HL60, P388, HT29, and MCF7 cell lines with the IC50 values ranging from 14.40 to 52.63 μM compared to the positive control compound (ellipticine) with the IC50 values ranging from 1.34 to 1.99 μM.

In-silico profiling, design, molecular docking computation, and drug kinetic model evaluation of novel curcumin derivatives as potential anticancer agents

The alarming trend of leukemia cell lines that are multidrug-resistant has prompted scientists to scramble for effective new anticancer treatments. Therefore, it remains an intriguing scientific task to optimize curcumin by trying to introduce molecular alteration to its vital structure to improve the biological effect against the P388 cell line or get around resistance phenomena. Regardless of the wide range of medications that are now being studied, Prednisone remains the most important and efficient part of chemotherapy that the WHO recommends. This article discusses the QSAR-oriented model and in silico assessment of some potent curcumin derivatives' anticancer activity against the P388 cell line. The solidity and propensity for prediction of the model were ensured by using stringent validation procedures. The activity of these derivatives was shown to be unrelated to lipophilicity, while shorter N-N distances and short substituents result in quite bioactive molecules. This information was used to design potent molecules that demonstrate good quality as per the assessment based on the Lead-Like Soft rule is acceptable for drug-like molecules. Also, molecule d2 does not possess any toxic effects risk alerts, suggesting drug-adherent conduct. While Prednisone the reference drug has a toxicity risk alert in red, suggesting non-adherent conduct for Prednisone. Hence, the novel molecules are promising anticancer agents.

An Insight of Cryptocarya Secondary Metabolites as Anticancer P388: Study of Molecular Docking and ADMET Properties

Secondary metabolites isolated from Cryptocarya was known to have various activity especially their cytotoxicity in P388 cell. There were two species of Cryptocarya studied in this research that were Cryptocarya konishii and Cryptocarya lucida. In both species, 8 isolate compounds had bioactivity as anticancer in P388 cells. This study aimed to know the binding affinity and ADMET properties of each isolated compound through P-glycoprotein substrate since this protein was reported to be responsible for the inhibition of P388 cells. Molecular docking was performed using AutoDock4 and AutoDockTools software to know the binding energy and interaction of isolate compounds against the P-glycoprotein substrate. ADMET properties calculation was done using the pkCSM web server for all compounds. Molecular docking results showed that Kurzichalcolactone B (7) isolated from C. lucida had the lowest binding energy. It resulted in the highest total intermolecular energy from the contribution of van der Waals and hydrogen bond energy. The lowest binding energy is indicating the stable interaction of ligand and substrate. Calculation of ADMET properties resulted that some of the isolate compounds fulfilling the minimum standard parameters in ADMET properties.

Two Onnamide Analogs from the Marine Sponge Theonella conica: Evaluation of Geometric Effects in the Polyene Systems on Biological Activity.

Two previously unreported onnamide analogs, 2Z- and 6Z-onnamides A (1 and 2), were isolated from the marine sponge Theonella conica collected at Amami-Oshima Is., Kagoshima Prefecture, Japan. Structures of compounds 1 and 2 were elucidated by spectral analysis. Structure-activity relationships (SARs) for effects on histone modifications and cytotoxicity against HeLa and P388 cells were characterized. The geometry in the polyene systems of onnamides affected the histone modification levels and cytotoxicity.

Species-specific secondary metabolism by actinomycetes of the genus Phytohabitans and discovery of new pyranonaphthoquinones and isatin derivatives.

To further exploit secondary metabolic potential of a minor actinomycete genus Phytohabitans within the family Micromonosporaceae, metabolite profiling by HPLC-UV analysis, combined with 16S rDNA sequence-based phylotyping were attempted on seven Phytohabitans strains available at the public culture collection. The strains were grouped into three clades and each exhibited unique and distinct metabolite profiles, which were highly conserved among strains within the same clade. These results were consistent with previous observations on two other actinomycetes genera, reconfirming species-specificity of secondary metabolite production, which were conventionally thought to be strain-specific. A strain RD003215, belonging to the P. suffuscus clade, produced multiple metabolites, some of which were presumed to be naphthoquinones. Liquid fermentation followed by chromatographic separation of the broth extract led to the discovery of three new pyranonaphthoquinones, designated habipyranoquinones A-C (1-3), and one new isatin derivative, (R)-N-methyl-3-hydroxy-5,6-dimethoxyoxindole (4), along with three known synthetic compounds, 6,8-dihydroxydehydro-α-lapachone (5), N-methyl-5,6-dimethoxyisatin (6), and 5,6-dimethoxyisatin (7). Structures of 1-4 were unequivocally elucidated by NMR, MS, and CD spectral analysis, with assistance of density functional theory-based NMR chemical shift prediction and ECD spectral calculation. Compound 2 displayed antibacterial activity against Kocuria rhizophila and Staphylococcus aureus with MIC 50 µg/mL and cytotoxicity against P388 murine leukemia cells with an IC50 value of 34 µM. Compounds 1 and 4 also showed cytotoxicity against P388 cells with IC50 values of 29 and 14 µM, respectively.

Deregulated expression of the 14q32 miRNA cluster in clear cell renal cancer cells.

Clear cell renal cell carcinomas (ccRCC) are characterized by arm-wide chromosomal alterations. Loss at 14q is associated with disease aggressiveness in ccRCC, which responds poorly to chemotherapeutics. The 14q locus contains one of the largest miRNA clusters in the human genome; however, little is known about the contribution of these miRNAs to ccRCC pathogenesis. In this regard, we investigated the expression pattern of selected miRNAs at the 14q32 locus in TCGA kidney tumors and in ccRCC cell lines. We demonstrated that the miRNA cluster is downregulated in ccRCC (and cell lines) as well as in papillary kidney tumors relative to normal kidney tissues (and primary renal proximal tubule epithelial (RPTEC) cells). We demonstrated that agents modulating expression of DNMT1 (e.g., 5-Aza-deoxycytidine) could modulate 14q32 miRNA expression in ccRCC cell lines. Lysophosphatidic acid (LPA, a lysophospholipid mediator elevated in ccRCC) not only increased labile iron content but also modulated expression of a 14q32 miRNA. Through an overexpression approach targeting a subset of 14q32 miRNAs (specifically at subcluster A: miR-431-5p, miR-432-5p, miR-127-3p, and miR-433-3p) in 769-P cells, we uncovered changes in cellular viability and claudin-1, a tight junction marker. A global proteomic approach was implemented using these miRNA overexpressing cell lines which uncovered ATXN2 as a highly downregulated target. Collectively, these findings support a contribution of miRNAs at 14q32 in ccRCC pathogenesis.