Prostate Adenocarcinoma Cell Line Research Articles

A bis-quinoline ruthenium(II) arene complex with submicromolar cytotoxicity in castration-resistant prostate cancer cells.

A new Ru(II) arene chlorido organometallic complex [(η6-p-cymene)(L)RuCl]PF6 (named as pCYRuL) using 2-bis(quinolin-2-ylmethylene) hydrazine (L) was developed that exhibits potent anticancer activity against castration-resistant prostate cancer (CRPC) (IC50 = 0.71 μM), and it is 45 times more effective than the standard drug cisplatin (IC50 = 31.3 μM) in a castration-resistant human prostatic adenocarcinoma cell line (PC-3) but non-toxic in normal human kidney cells (HK2) as well as normal breast cells (MCF10A) and found that pCYRuL exerted anticancer activity via apoptosis induction and cell cycle arrest in the G2/M phase of PC-3 cells.

Chemical composition, antioxidant, antimicrobial and antiproliferative activities of the leaf essential oil of Roxburgh’s Cherry (Eugenia roxburghii), an underutilized species

Eugenia roxburghii (Myrtaceae), known as Roxburgh’s Cherry, is a wild edible fruit-producing plant having multiple use in traditional and folk medicines in India and Sri Lanka. In spite of its nutritional and medicinal importance, there is no report on the chemical composition and bioactivities of the essential oil (EO) of this species. In this article, the chemical composition of the leaf EO of E. roxburghii has been reported for the first time along with its antioxidant, antimicrobial and antiproliferative activities. GC–MS analysis of EO led to the identification of 41 compounds with β-caryophyllene (22.32%) as the major constituent. The EO exhibited strong antioxidant capacity in DPPH and ABTS assays with IC50 values of 15.12 ± 0.23 μg/mL and 7.32 ± 0.05 μg/mL, respectively. It also exhibited strong reducing power ability (EC50 = 1.47 ± 0.06 μg/mL). The EO showed highest sensitivity against enteric pathogenic bacteria, Shigella flexneri (ZOI = 24 ± 0.4 mm, MIC = 31.25 μg/mL) validating its ethnomedicinal claim against dysentery. Further, E. roxburghii EO showed significant antiproliferative activity against human colon (HCT-116, IC50 = 64.84 μg/mL) and prostatic adenocarcinoma (PC-3, IC50=70.11 μg/mL) cell lines. The present study suggests that EO can be used as an antiproliferative agent and for the treatment of enteric diseases.

Antiproliferative and Apoptosis Effects of Hybrid Varieties of Vitis vinifera L. Sweet Sapphire and Sweet Surprise on Human Prostate Cancer Cells Using In Vitro and In Silico Approaches.

Grape hybrids are characterized by different chemical compositions; often with high hybrids are characterized by different chemical compositions, often with a high phenolic content and a specific profile of anthocyanins. The aim of study was to characterize the constituents of hybrid Vitis vinifera L. varieties Sweet Sapphire (SA) and Sweet Surprise (SU) extracts and their influence on apoptosis induction and antiproliferative effects on human prostate cancer cells. We used the MTT assay to evaluate the cytotoxic effect of extracts of SA and SU, on the prostate adenocarcinoma cell lines PC-3 and DU-145. To analyze the inhibiting impact by flow cytometry, used 24 and 48 hours. Anthocyanins were quantified by liquid chromatography and analysed by their absorption rate, hepatotoxicity, blood concentration, blood-brain barrier passage ability and maximum recommended dose by in silico approaches. Our results showed that malvidin derivatives present the highest content in both cultivars. We identified 14.46mg/100g malvidin-3-O-glycoside in SA and 2.76 mg/100 g in SU. A reduction in cell viability of DU-145 (45 and 65%) and PC-3 (63 and 67%) cells after 48h treatment with SA and SU, respectively, was found via MTT assay. Flow cytometry showed that the treatment with extracts from SA and SU had an inhibitory impact on cell development due to G2/M arrest and caused a rise in apoptotic cells compared to control group. None of the anthocyanin presented hepatotoxicity as well as blood-brain barrier passage ability. Peonidin 3-O-glucoside had the lower maximum recommended dose as well as the highest intestinal absorption rate. However, delphinidin 3-O-glucoside had the highest blood concentration values. The findings of this study highlight the potential of hybrid Vitis vinifera L. varieties as an important source of natural antioxidants and their protective effect against prostate cancer cells as well as elucidate in part their anthocyanin's metabolism.

Design and Characterization of pMyc/pMax Peptide-Coupled Gold Nanosystems for Targeting Myc in Prostate Cancer Cell Lines.

Myc and Max are essential proteins in the development of prostate cancer. They act by dimerizing and binding to E-box sequences. Disrupting the Myc:Max heterodimer interaction or its binding to E-box sequences to interrupt gene transcription represent promising strategies for treating cancer. We designed novel pMyc and pMax peptides from reference sequences, and we evaluated their ability to bind specifically to E-box sequences using an electrophoretic mobility shift assay (EMSA). Then, we assembled nanosystems (NSs) by coupling pMyc and pMax peptides to AuNPs, and determined peptide conjugation using UV-Vis spectroscopy. After that, we characterized the NS to obtain the nanoparticle's size, hydrodynamic diameter, and zeta potential. Finally, we evaluated hemocompatibility and cytotoxic effects in three different prostate adenocarcinoma cell lines (LNCaP, PC-3, and DU145) and a non-cancerous cell line (Vero CCL-81). EMSA results suggests peptide-nucleic acid interactions between the pMyc:pMax dimer and the E-box. The hemolysis test showed little hemolytic activity for the NS at the concentrations (5, 0.5, and 0.05 ng/µL) we evaluated. Cell viability assays showed NS cytotoxicity. Overall, results suggest that the NS with pMyc and pMax peptides might be suitable for further research regarding Myc-driven prostate adenocarcinomas.

Technical report: liquid overlay technique allows the generation of homogeneous osteosarcoma, glioblastoma, lung and prostate adenocarcinoma spheroids that can be used for drug cytotoxicity measurements.

Introduction: The mechanisms involved in cancer initiation, progression, drug resistance, and disease recurrence are traditionally investigated through in vitro adherent monolayer (2D) cell models. However, solid malignant tumor growth is characterized by progression in three dimensions (3D), and an increasing amount of evidence suggests that 3D culture models, such as spheroids, are suitable for mimicking cancer development. The aim of this report was to reaffirm the relevance of simpler 3D culture methods to produce highly reproducible spheroids, especially in the context of drug cytotoxicity measurements. Methods: Human A549 lung adenocarcinoma, LnCaP prostate adenocarcinoma, MNNG/HOS osteosarcoma and U251 glioblastoma cell lines were grown into spheroids for 20days using either Liquid Overlay Technique (LOT) or Hanging Drop (HD) in various culture plates. Their morphology was examined by microscopy. Sensitivity to doxorubicin was compared between MNNG/HOS cells grown in 2D and 3D. Results: For all cell lines studied, the morphology of spheroids generated in round-bottom multiwell plates was more repeatable than that of those generated in flat-bottom multiwell plates. HD had no significant advantage over LOT when the spheroids were cultured in round-bottom plates. Finally, the IC50 of doxorubicin on MNNG/HOS cultured in 3D was 18.8 times higher than in 2D cultures (3D IC50 = 15.07 ± 0.3µM; 2D IC50 = 0.8 ± 0.4µM; *p < 0.05). Discussion: In conclusion, we propose that the LOT method, despite and because of its simplicity, is a relevant 3D model for drug response measurements that could be scaled up for high throughput screening.

Green Synthesis of Biocompatible Hybrid Ginger/Chitosan Carbon Nanodot Exhibiting Antiproliferative Activity on Carcinoma Cells

Carbon Nanodots (CDs)-modified chitosan and ginger (ginger/chitosan@CD) based biocompatible substrate was built with the purpose of assessing antiproliferation effect on prostate cancer cell line (PC-3), human prostate adenocarcinoma cell line (LNCaP), and breast cancer cell line (MCF-7). CDs were fabricated through a solvothermal synhesis process, and characterized with TEM, SERS, and UV-vis spectroscopy. In this study, the cytotoxic impact of ginger/chitosan CD, which was synthesized for the first time, was evaluated as a function of both dose (maximum concentration 500 μg/mL) and time (in 24 and 48 hours) in cancer cell lines by XTT assay. After 48 hours, the ginger/chitosan@CD combination was found to have a 50% inhibitory concentration (IC50) of 178.08 μg/mL in the PC-3 cell line, 246.44 μg/mL in the LNCaP prostate cancer cells, and 345.74 μg/mL in the MCF-7. Cancer cell proliferation was efficiently suppressed by the ginger/chitosan@CDs.

Exportin 1 inhibition prevents neuroendocrine transformation through SOX2 down-regulation in lung and prostate cancers.

In lung and prostate adenocarcinomas, neuroendocrine (NE) transformation to an aggressive derivative resembling small cell lung cancer (SCLC) is associated with poor prognosis. We previously described dependency of SCLC on the nuclear transporter exportin 1. Here, we explored the role of exportin 1 in NE transformation. We observed up-regulated exportin 1 in lung and prostate pretransformation adenocarcinomas. Exportin 1 was up-regulated after genetic inactivation of TP53 and RB1 in lung and prostate adenocarcinoma cell lines, accompanied by increased sensitivity to the exportin 1 inhibitor selinexor in vitro. Exportin 1 inhibition prevented NE transformation in different TP53/RB1-inactivated prostate adenocarcinoma xenograft models that acquire NE features upon treatment with the aromatase inhibitor enzalutamide and extended response to the EGFR inhibitor osimertinib in a lung cancer transformation patient-derived xenograft (PDX) model exhibiting combined adenocarcinoma/SCLC histology. Ectopic SOX2 expression restored the enzalutamide-promoted NE phenotype on adenocarcinoma-to-NE transformation xenograft models despite selinexor treatment. Selinexor sensitized NE-transformed lung and prostate small cell carcinoma PDXs to standard cytotoxics. Together, these data nominate exportin 1 inhibition as a potential therapeutic target to constrain lineage plasticity and prevent or treat NE transformation in lung and prostate adenocarcinoma.

ZERDEÇALIN BİYOAKTİF BİLEŞİĞİ KURKUMİN, GEMSİTABİNİN PROSTAT KANSERİ HÜCRELERİNDEKİ ANTİ-MALİGNANT ÖZELLİĞİNİ GELİŞTİREBİLİR

Objective: The aim of this study was to investigate the possible synergistic effect of curcumin on the anticancer features of gemcitabine on prostate cancer cells. Material and Method: The human prostate adenocarcinoma cell line LNCaP was used in the studies. The effect of the co-administration of gemcitabine and curcumin on the viability of LNCaP cells was investigated by the WST-1 assay. Autophagy, ubiquitin-proteasome system (UPS), unfolded protein response (UPR) and cell death-associated proteins, androgenic signaling, proto-oncogenic, angiogenic and epithelial-mesenchymal transition (EMT) associated protein levels were investigated by immunoblotting studies. Result and Discussion: Our results showed that curcumin potentiated the anticancer effects of gemcitabine on LNCaP cells. Co-administration of curcumin and gemcitabine strengthened the suppressive effect of gemcitabine on cell viability. Moreover, co-administration modulated the autophagy, more strongly stimulated UPS and UPR, suppressed androgenic signaling, led to the activation of cell death-related poly [ADP-ribose] polymerase 1 (PARP-1) and caspase-3 and strongly suppressed the expression levels of proto-oncogenic c-Myc and angiogenic vascular endothelial growth factor-A (VEGF-A). In addition, it was determined that co-administration negatively regulated EMT by stimulating E-cadherin expression and suppressing N-cadherin level. These results suggest that the combined usage of gemcitabine and curcumin may offer a potent therapeutic approach to prostate cancer by enhancing the anticancer effects of gemcitabine.

Abstract B017: Lineage-specific PRC2 targets and response to EZH2 inhibition in neuroendocrine prostate cancer

Abstract Lineage plasticity is an emerging mechanism of treatment-resistance in prostate cancer, which can manifest clinically as histologic transformation from prostate adenocarcinoma to small-cell neuroendocrine prostate cancer (NEPC). NEPC is associated with poor prognosis, and new treatment strategies are urgently needed. Epigenetic dysregulation has been implicated as a key driver of prostate cancer lineage plasticity, mediated in part by EZH2. EZH2 is a component of the polycomb repressive complex (PRC2) regulating the transcriptional repressive mark H3K27me3. EZH2 also controls cell-fate determination during normal development via bivalent promoters bearing both repressive H3K27me3 and active H3K4me3 marks. EZH2 is targetable, and several EZH2 inhibitors are in clinical development; these trials are currently in biomarker-unselected prostate cancer populations. A thorough understanding of the mechanism of action of EZH2 is imperative for predicting optimal response to EZH2i and for the development of robust combination treatment strategies. Cell viability studies across a diverse panel of prostate cancer models representing adenocarcinoma and NEPC indicated variable responses to tazemetostat (EZH2i) treatment. The castration-resistant prostate adenocarcinoma (CRPC) cell line LNCaP-abl showed a greater than 50% reduction in cell viability upon EZH2i (5uM) while LNCaP and 22Rv1 showed moderate response with 15-25% reduction in cell viability. However, none of our 5 NEPC patient-derived models showed significant response to EZH2i which was also validated in vivo. Hierarchical clustering of RNA-seq profiles of responder and non-responder models revealed distinct cell-type specific EZH2 targets and clusters of genes that may be associated with response to EZH2i. Approximately 15% of the EZH2 targets in the NEPC organoid WCM154 carried bivalency in their promoters including neuronal lineage determining genes such as ASCL1 and LHX2. Gene set enrichment analysis of EZH2 targets bearing bivalency in NEPC revealed association with neurogenesis and cell proliferation pathways. Further upregulation of neuronal-lineage PRC2 targets in NEPC models upon EZH2i may provide justification for lack of lineage reversal and maintenance of terminal differentiation. While there is evidence that EZH2i leads to lineage reversion in adeno-to-NEPC transitioning preclinical models, our data shows that EZH2i was associated with modest response in NEPC models and did not revert lineage plasticity. Our results provide insights into a potential role for bivalent promoters in EZH2-mediated lineage reprogramming and sustenance of terminal differentiation upon EZH2i in NEPC. Understanding lineage-specific action of EZH2i may inform biomarkers in ongoing clinical trials. Genes and pathways dysregulated because of their bivalency might also be exploited as potential candidates to be co-targeted with EZH2i. Citation Format: Varadha Balaji Venkadakrishnan, Richa Singh, Yasutaka Yamada, Kei Mizuno, Adam G. Presser, Sheng-Yu Ku, Henry Long, David Rickman, Himisha Beltran. Lineage-specific PRC2 targets and response to EZH2 inhibition in neuroendocrine prostate cancer [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr B017.

Comprehension characterization of prostate cancer cell lines identified JAK-STAT3 signaling in lineage-switched non-neuroendocrine cells.

e17005 Background: Prostate cancer (PCa) is the most diagnosed cancer type among American men. Androgen receptor (AR) targeted therapy as the gold standard of PCa has been challenged by treatment resistance. Lineage plasticity enables PCa cells to transit from prostate luminal epithelia to alternative lineages to bypass their dependency on AR signaling. Previously we established prostatic HOX code to study the prostatic lineage. We found that cells with neuroendocrine (NE) phenotype have a lineage switch. Besides, a subgroup of AR negative, NE negative samples also display lineage switch, among which is DU145, a human PCa cell line. Methods: RNA-seq data of PCa cell lines were obtained from GEO then aligned and quantified using human GRCh38 genome with STAR-Salmon pipeline. Downstream differential expression and enrichment analysis was performed with R. Immunohistochemistry (IHC) was used to assess protein levels on cell line derived xenografts. Results: 45 RNA-seq datasets of 6 prostate adenocarcinoma cell lines (VCaP, LNCaP, C4-2, 22RV1, PC3 and DU145) and 8 RNA-seq datasets of neuroendocrine prostate cancer cell line (H660) passed quality control. Principle component analysis indicates that samples from the same cell line had similar transcriptomes. Differential expression analysis followed by gene function enrichments analysis identified the marker genes and key signaling pathways of each cell line. HOX code analysis confirmed previous conclusion that all PCa cell lines but DU145 and H660 samples maintained prostatic HOX code. IHC staining on cell line-derived xenografts confirmed the protein expression of selected marker genes (TUBB2B for H660, HGF for VCaP and 22RV1, CXCL1 and KRT7 for DU145 and PC3). The three AR-negative PCa cell lines were further characterized in terms of AR/ NE activity scores, transcriptome identities, HOX codes. These analyses demonstrated that PC3 is a prostate progenitor like cell line, H660 is a lineage-switched NE cell line, and DU145 is a lineage-switched non-NE cell line. Further, signaling enrichment analysis revealed that DU145 cells had higher JAK-STAT3 signaling activity compared to other cell lines. IHC staining detected strong p-STAT3 signals on DU145-derived tumors but not on any other cell line-derived tumors. Conclusions: Human PCa cell lines were fully characterized based on their transcriptomes with high sample numbers. PC3 represents cells dedifferentiated into progenitor stage. DU145 and H660 represent cells that further trans-differentiate into alternative lineages. Neuron master regulators drive H660 cells into NE lineage and STAT3 drives DU145 cells into a non-NE lineage. DU145, representing a non-NE PCa class that have lost prostatic lineage, would be a suitable model system to study JAK-STAT targeting therapy.

Gleason Score-related MT1L as biomarker for prognosis in prostate adenocarcinoma and contribute to tumor progression in vitro.

The Gleason Score is well correlated with biological behavior and prognosis in prostate adenocarcinoma (PRAD). This study was derived to determine the clinical significance and function of Gleason-Score-related genes in PRAD. RNA-sequencing profiles and clinical data were extracted from the The Cancer Genome Atlas PRAD database. The Gleason-Score-related genes were screened out by the Jonckheere-Terpstra rank-based test. The "limma" R package was performed for differentially expressed genes. Next, a Kaplan-Meier survival analysis was performed. Correlation MT1L expression levels with tumor stage, non-tumor tissue stage, radiation therapy, and residual tumor were analyzed. Further, MT1L expression was detected in PRAD cell lines by reverse transcription-quantitative polymerase chain reaction assay. Overexpression of MT1L was constructed and used for cell count kit-8, flow cytometric assay, transwell assay, and wound-healing assay. Survival analysis showed 15 Gleason-Score-related genes as prognostic biomarkers in PRAD. The high-frequency deletion of MT1L was verified in PRAD. Furthermore, MT1L expression was decreased in PRAD cell lines than RWPE-1 cells, and overexpression of MT1L repressed cell proliferation and migration, and induced apoptosis in PC-3 cells. Gleason-Score-related MT1L may serve as a biomarker of poor prognostic biomarker in PRAD. In addition, MT1L plays a tumor suppressor in PRAD progression, which is beneficial for PRAD diagnosis and treatment research.

MDR1 Inhibition Reverses Doxorubicin-Resistance in Six Doxorubicin-Resistant Canine Prostate and Bladder Cancer Cell Lines.

Acquired chemoresistance during chemotherapy, often accompanied by cross- and multi-resistance, limits therapeutic outcomes and leads to recurrence. In order to create in vitro model systems to understand acquired doxorubicin-resistance, we generated doxorubicin-resistant sublines of canine prostate adenocarcinoma and urothelial cell carcinoma cell lines. Chemoresistance to doxorubicin, cross-resistance to carboplatin, and the reversibility of the acquired resistance by the specific MDR1-inhibitor tariquidar were quantified in metabolic assays. Resistance mechanisms were characterized by expression of the efflux transporters MDR1 and RALBP1, as well as the molecular target of doxorubicin, TOP2A, with qPCR and Western blotting. Six out of nine cell lines established stable resistance to 2 µM doxorubicin. Drug efflux via massive MDR1 overexpression was identified as common, driving resistance mechanism in all sublines. MDR1 inhibition with tariquidar extensively reduced or reversed the acquired, and also partly the parental resistance. Three cell lines developed additional, non-MDR1-dependent resistance. RALBP1 was upregulated in one resistant subline at the protein level, while TOP2A expression was not altered. Combination therapies aiming to inhibit MDR1 activity can now be screened for synergistic effects using our resistant sublines. Nevertheless, detailed resistance mechanisms and maintained molecular target expression in the resistant sublines are still to be examined.

New sulfa drug derivatives and their zinc(II) complexes: synthesis, spectroscopic properties and in vitro cytotoxic activities

Synthesis of four sulfa drug derivatives (L1–L4 ) and Zn(II) complexes derived from sulfonamide group antibiotic substances was carried out using the hydrothermal technique (HT) and their structures of the obtained compounds were explained using elemental analysis (EA), FT-IR and NMR (1H- and 13C-). Cytotoxic activities of four novel sulfa drug based-Schiff base compounds and their Zn(II) complexes were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay using MCF-7 (human breast cancer), Caco-2 (human colorectal adenocarcinoma), A2780 (human ovarian cancer) and LNCaP (human prostate adenocarcinoma) cell lines. LogIC50 values of all obtained compounds were computed with the Graphpad Prism 6 program after 24 h of treatment for MCF-7, Caco-2, A2780 and LNCaP cells. Comet assay experiments were performed using LogIC50 concentrations of all compounds to determine DNA damage. Based on the data obtained, all compounds significantly decreased MCF-7, Caco-2, A2780 and LNCaP cell viability compared to the control groups (p < 0.05).

Tail-Approach-Based Design and Synthesis of Coumarin-Monoterpenes as Carbonic Anhydrase Inhibitors and Anticancer Agents.

In this study, sixty novel coumarin-monoterpene compounds were synthesized in two series [thirty-two compounds (12-43) bearing a triazole ring in the first series, and twenty-eight compounds (44-71) bearing an alkyl chain in the second one]. Their inhibitory effects on the human carbonic anhydrase (hCA) isoforms I, II, IX, and XII and anticancer potentials were determined. All synthesized molecules selectively inhibited CA IX and XII. 23 and 42 were found to be the strongest inhibitors, with K i values of 1.9 nM against hCA IX. Also, 70 showed the highest inhibitory activity with a K i value of 4.9 nM against hCA XII. Moreover, their cytotoxic effects on colon adenocarcinoma (HT-29), prostate adenocarcinoma (PC-3), and breast adenocarcinoma (MCF-7) cell lines were evaluated. According to the cytotoxicity results, 14 (IC50 = 2.48 μM) and 63 (IC50 = 3.91 μM) exhibited the highest cytotoxicity on the MCF-7 cells, while 23 showed the strongest cytotoxic effect on both PC-3 (IC50 = 9.40 μM) and HT-29 (IC50 = 12.10 μM) cell lines. 14, 23, and 66 decreased CA IX and CA XII protein expression in HT-29 cells, while 23 and 66 showed the strongest reduction of both CA IX and CA XII in MCF-7 cells. All of the selected compounds increased total apoptosis in a concentration-dependent manner in HT-29 and MCF-7 cells. 14 has the strongest apoptotic effect in MCF-7 cells. 23 increased early apoptosis primarily, while 14 and 66 increased total apoptosis in HT-29. In addition, PI/Hoechst staining proves that apoptotic cells are increased in HT-29 with an effect of 14, 23, and 66. As a result of the modeling studies, it has been shown that only the open coumarin form of the compounds can interact directly with the active-site Zn2+ ion. It has been shown that coumarin-monoterpene structures with different alkyl and monoterpene groups both specifically inhibit CA IX and XII and exhibit specific cytotoxicity in different cell lines.

Monometallic complexes of novel dihydrazone‐oxime ligand: Synthesis, structure, spectral, DFT, and in vitro bioactivity investigation against fungal, bacterial, and tumor cells

In the current study, uni‐metallic Cu2+, Ni2+, Mn2+, Co2+, and Zn2+ complexes of ONN‐tridentate dihydrazone‐oxime incorporating indolin‐2‐one moiety (HL,1) were synthesized and structurally characterized employing elemental, spectral, thermo‐gravimetric, magnetic, and molar conductivity measurements, to investigate the structure and composition of these new uni‐metallic complexes. According to the results of several analyses, the dihydrazone‐oxime behaved as a mono‐negative or neutral tridentate ONN chelator bound the metal cations in molar ratio (1M:1L) through the isatinic carbonylic oxygen, azomethine nitrogen, and nitrogen atom of deprotonated/protonated oximatic moiety forming uni‐metallic complexes with formulae [M(L)XYZ(H2O)].nH2O (where M = Cu2+ (2–4), Ni2+, Mn2+, Co2+, or Zn2+, X = Cl, CH3COO, or NO3, Y = H2O or 0, Z = H2O or 0, n = 2, 0) and [Cu (HL)(SO4)(H2O)]. The Cu2+ complexes were found to have a tetragonally distorted octahedral or square pyramidal geometry, whereas those of Mn2+, and Ni2+, exhibited a tetrahedral geometry but Co2+ complex demonstrated a distorted octahedral geometry. Based on DFT‐B3LYP technique and 6‐311G++(d,p) basis set, The DFT data confirmed the structural investigation of dihydrazone‐oxime and its complexes, revealing that the complexes reactivity is more than un‐complexed dihydrazone‐oxime. Moreover, quantum chemical parameters of dihydrazone‐oxime and its uni‐metallic complexes like the energies of both lowest unoccupied molecular orbital (ELUMO) and highest occupied molecular orbital (EHOMO), separation energy (ΔE = ELUMO − EHOMO), chemical potential, absolute electronegativity, absolute hardness, and absolute softness were obtained. According to the thermogravimetric analysis, the complexes breakdown in two to four disintegrating phases in range of temperature 25°C to 575°C leaving the oxide of metal or metal oxide contaminated with carbon. In vitro microbicide effect of the un‐complexed dihydrazone‐oxime and its uni‐metallic complexes toward various bacterial and fungal strains was tested by agar‐well diffusion method. It is demonstrated that the complexes are more effective than free dihydrazone‐oxime. The antitumor activity has been also analyzed against three cell lines including prostate adenocarcinoma (PC‐3), ovarian adenocarcinoma (SKOV3), and cervical cancer (HeLa) cell lines. Studies showed that all complexes have greater inhibition activity against the above three cell lines than the free dihydrazone oxime and the most effective compound was the Zn2+ complex (9) with IC50, 27.42, 35.16, and 46.77 μg/ml, respectively.

Coumarinolignoid and Indole Alkaloids from the Roots of the Hybrid Plant Citrus × paradisi Macfad (Rutaceae).

A phytochemical investigation of the roots of Citrus × paradisi Macfad. (Rutaceae) led to the isolation of two new compounds, namely 1-formyl-5-hydroxy-N-methylindolin-1-ium (1) and decyloxycleomiscosin D (2), along with ten known compounds: 1,1-dimethylpyrrolidin-1-ium-2-carboxylate (3), furan-2,3-diol (4), 5-methoxyseselin (5), umbelliferone (6), scopoletin (7), citracridone I (8), citracridone II (9), citracridone III (10), limonin (11) and lupeol (12). The structures were determined through the comprehensive spectroscopic analysis of 1D and 2D NMR and EI- and ESI-MS, as well as a comparison with the published data. Notably, compounds 3 and 4 from the genus Citrus are reported here for the first time. In addition, the MeOH extract of the roots and compounds 1-7 were screened against the human adenocarcinoma alveolar basal epithelial cell line A549 and the Caucasian prostate adenocarcinoma cell line PC3 using the MTT assay. While the extract showed significant activity, with IC50 values of 35.2 and 38.1 µg/mL, respectively, compounds 1-7 showed weak activity, with IC50 values of 99.2 to 250.2 µM and 99.5 to 192.7 µM, respectively.

Antiproliferative Action of Methanolic Petiole Extract of Eichhornia Crassipes on Human Prostate Adenocarcinoma Cell Line: An In Vitro Study.

An increasing number of people are turning to herbal medicines in their search for innovative pharmaceuticals since they are effective treatments for a wide variety of conditions and traditional herbs are rich in bioactive chemicals. In this study, we looked at whether or not a petiole extract of Eichhornia crassipes preserved in methanol inhibited the proliferation of prostate cancer (PC3) cell lines. Lakes in Ezhikkara, Ernakulum, Kerala, were the source of E. crassipes. Soxhlet extraction was used to create the extract. 3-[4,5-dimethylthiazol-2-yl]2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the cell viability of methanolic petiole extract at various concentrations. Mean and standard deviation was used to determine absorbance scores. Utilizing probit analysis, we determined the IC50 value. The descriptive statistics to measure the percent of viable cells along with the regression equation were calculated using SPSS. It has been shown that the methanol extract significantly impacted PC3 cell lines' capacity to survive. It was also determined that increasing the medication concentration resulted in a decrease in cell viability. The percentage of living cells was measured after being exposed to methanol extracts at concentrations of 12.5 μg/ml, 25 μg/ml, 50 μg/ml, 100 μg/ml, and 200 μg/ml, and found to be 95.13, 85.88, 76.12, 64.33, and 53.62 percent, respectively. With IC50 values of 199.488 g/ml, it was shown that methanolic petiole extracts of E. crassipes are cytotoxic. Using probit analysis, we determined that the regression equation is y = -0.2051x + 90.915, with an R2 value of 0.893. As a result of its chemotherapeutic properties, the E. crassipes petiole extract has the potential to be employed in therapeutic applications, with the ultimate goal of bettering prostate cancer management practices and clinical results by drastically lowering cell viability. The study's results may pave the way for fresh chemotherapeutic approaches to be developed for the treatment of androgen-independent prostate cancer.

Unraveling the Metabolic Alterations Induced by Zika Infection in Prostate Epithelial (PNT1a) and Adenocarcinoma (PC-3) Cell Lines.

The outbreak of Zika virus infection in 2016 led to the identification of its presence in several types of biofluids, including semen. Later discoveries associated Zika infection with sexual transmission and persistent replication in cells of the male reproductive tract. Prostate epithelial and carcinoma cells are favorable to virus replication, with studies pointing to transcriptomics alterations of immune and inflammation genes upon persistence. However, metabolome alterations promoted by the Zika virus in prostate cells are unknown. Given its chronic effects and oncolytic potential, we aim to investigate the metabolic alterations induced by the Zika virus in prostate epithelial (PNT1a) and adenocarcinoma (PC-3) cells using an untargeted metabolomics approach and high-resolution mass spectrometry. PNT1a cells were viable up to 15 days post ZIKV infection, in contrast to its antiproliferative effect in the PC-3 cell lineage. Remarkable alterations in the PNT1a cell metabolism were observed upon infection, especially regarding glycerolipids, fatty acids, and acylcarnitines, which could be related to viral cellular resource exploitation, in addition to the over-time increase in oxidative stress metabolites associated with carcinogenesis. The upregulation of FA20:5 at 5 dpi in PC-3 cells corroborates the antiproliferative effect observed since this metabolite was previously reported to induce PC-3 cell death. Overall, Zika virus promotes extensive lipid alterations on both PNT1a and PC-3 cells, promoting different outcomes based on the cellular metabolic state.

Cyc-DEP: Cyclic immunofluorescence profiling of particles collected using dielectrophoresis.

Cancer is a highly heterogenous disease that requires precise detection tools and active surveillance methods. Liquid biopsy assays provide an agnostic way to follow the complex trajectory of cancer, providing better patient stratification tools for optimized treatment. Here, we present the development of a low-volume liquid biopsy assay called cyc-DEP (cyclic immunofluorescent imaging on dielectrophoretic chip) to profile biomarkers collected on a dielectrophoretic microfluidic chip platform. To enable on-chip cyclic imaging, we optimized a fluorophore quenching method and sequential rounds of on-chip staining with fluorescently conjugated primary antibodies. cyc-DEP allows for the quantification of a multiplex array of proteins using 25µl of a patient plasma sample. We utilized nanoparticles from a prostate adenocarcinoma (LNCaP) cell line and a panel of six target proteins to develop our proof-of-concept technique. We then used cyc-DEP to quantify blood plasma levels of target proteins from healthy individuals, low-grade and high-grade prostate cancer patients (n=3 each) in order to demonstrate that our platform is suitable for liquid biopsy analysis in its present form. To ensure accurate quantification of signal intensities and comparisons between different samples, we incorporated a signal intensity normalization method (fluorescent beads) and a custom signal intensity quantification algorithm that account for the distribution of signal across hundreds of collection regions on each chip. Our technique enabled a threefold improvement in multiplicity for detecting proteins associated with fluid samples, opening doors for early detection, and active surveillance through quantification of a multiplex array of biomarkers from low-volume liquid biopsies.

Abstract 2411: Di-ganglioside GD2 expression and role in promoting tumorigenicity in prostate cancer

Abstract Background &amp; Significance: Prostate cancer (PCa) is the second leading cause of cancer deaths (~34,000 in 2021 (ACS)) in American men. Castration resistance and resistance to the next-gen androgen receptor (AR) targeted drugs are major challenges. Castration resistance involves multiple mechanisms, including androgen-independent signaling by androgen receptor (AR)- or its variants, and lineage plasticity (LP) with AR-indifferent neuroendocrine (NE) differentiation. Identifying new vulnerabilities across these multitude mechanisms could provide new therapeutic avenues against castration-resistant PCa (CRPC). The cell-surface di-ganglioside GD2 is overexpressed in neural crest cell tumors such as neuroblastoma &amp; melanoma and chimeric (Dinutuximab) or humanized (Naxitamab) anti-GD2 antibodies are now FDA-approved for high-risk neuroblastoma therapy. GD2 expression is reported in other cancers such as breast cancer and glioma and is linked to cancer stem cell behavior. While limited prior studies have detected GD2 expression in PCa cell lines or tumor tissues, nothing is known about the functional role of GD2 in PCa. Objectives: We hypothesized that GD2 overexpression in PCa could play a pro-tumorigenic role and that linkage of GD2 overexpression with CRPC progression may reveal the potential of targeting GD2 for CRPC therapy. Study Design &amp; Results: Immunohistochemical analysis of PCa patient and patient-derived xenograft tissue microarrays (TMAs) revealed GD2 expression in a subset of tumor cells. Fluorescence-activated cell sorter analysis of PCa cell lines showed strong constitutive GD2 expression on murine CRPC cell line RM-1 (derived from mutant Ras and c-Myc overexpressing prostatic epithelial cells) and human PCa line 22Rv1 (overexpresses wild-type AR and ARv7 splice variant). GD2 expression was induced de novo upon induction of lineage plasticity in GD2-negative LNCaP C4-2 prostate adenocarcinoma cell line by shRNA knockdown (KD) of RB1 or TP53. High GD2 expression was also induced when C4-2B cells were made enzalutamide resistant (C4-2BER). Induction of GD2 expression correlated with increased expression of rate-limiting GD2 biosynthetic pathway enzyme GD3 synthase (GD3S). CRISPR-Cas9 mediated stable GD3S knockout (KO) in the RM1 cell line led to the loss of GD2 expression. The GD3S-KO RM1 cells exhibited reduced proliferation, migration, invasion, and tumor sphere forming ability compared to the control cells. Intratibial injections in castrated male C57BL/6 mice showed a significant reduction in tumor development by GD3S KO RM1 cells compared to control cells. Conclusions: Our studies demonstrate that GD2 is expressed in a subset of prostate cancers. Cell line-based studies show that GD2 expression promotes pro-tumorigenic traits. Future studies will assess the biological roles of GD2 in PCa and the potential of targeting GD2+ CRPC with antibody-based approved therapeutic agents. Citation Format: Aaqib M. Bhat, Bhopal C. Mohapatra, Insha Mushtaq, Sukanya Chakraborty, Samikshan Dutta, Sameer Mirza, Matthew D. Storck, Subodh M. Lele, Ming-Fong Lin, Bruce J. Trock, Karen S. Sfanos, Colm Morrissey, Eva Corey, Jonathan Melamed, Leah Cook, Kaustubh Datta, Jane Meza, Jawed Siddiqui, Surinder K. Batra, Vimla Band, Hamid Band. Di-ganglioside GD2 expression and role in promoting tumorigenicity in prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2411.