Colon Carcinoma Cell Line HCT116 Research Articles

Efficient Green Synthesis of Hydrazide Derivatives Using L-Proline: Structural Characterization, Anticancer Activity, and Molecular Docking Studies

Green synthesis using L-proline as an organocatalyst is crucial due to its reusability, mild conditions, clean reactions, easy workup, high purity, short reaction times, and high yields. However, existing methods often involve harsh conditions and longer reaction times. In this study, 2-cyano-N’-(2-cyanoacetyl)acetohydrazide (3) was prepared and condensed with various benzaldehyde derivatives to yield 2-cyano-N’-(2-cyano-3-phenylacryloyl)-3-phenylacrylohydrazide derivatives (5a–e, 7a,b) using a grinding technique with moist L-proline. Additionally, three 2-cyano-N’-(2-cyano-3-heterylbut-2-enoyl)-3-heterylbut-2-enehydrazides (9, 11, 13) were synthesized by condensing compound 3 with respective (heteraryl)ketones (8, 10, 12) following the same method. The synthesized compounds were characterized using IR, NMR, and MS spectroscopy. L-proline’s reusability was confirmed for up to four cycles without significant yield loss, showcasing the protocol’s efficiency and sustainability. The new compounds were screened for anticancer activities against the HCT-116 colon carcinoma cell line using the MTT assay. Molecular docking studies revealed the binding conformations of the most potent compounds to the target protein (PDB ID 6MTU), correlating well with in vitro results. In silico ADMET analysis indicated favorable pharmacokinetic properties, highlighting these novel compounds as promising targeted anti-colon cancer agents.

Memory effects of prior subculture may impact the quality of multiomic perturbation profiles

Mass spectrometry-based omics technologies are increasingly used in perturbation studies to map drug effects to biological pathways by identifying significant molecular events. Significance is influenced by fold change and variation of each molecular parameter, but also by multiple testing corrections. While the fold change is largely determined by the biological system, the variation is determined by experimental workflows. Here, it is shown that memory effects of prior subculture can influence the variation of perturbation profiles using the two colon carcinoma cell lines SW480 and HCT116. These memory effects are largely driven by differences in growth states that persist into the perturbation experiment. In SW480 cells, memory effects combined with moderate treatment effects amplify the variation in multiple omics levels, including eicosadomics, proteomics, and phosphoproteomics. With stronger treatment effects, the memory effect was less pronounced, as demonstrated in HCT116 cells. Subculture homogeneity was controlled by real-time monitoring of cell growth. Controlled homogeneous subculture resulted in a perturbation network of 321 causal conjectures based on combined proteomic and phosphoproteomic data, compared to only 58 causal conjectures without controlling subculture homogeneity in SW480 cells. Some cellular responses and regulatory events were identified that extend the mode of action of arsenic trioxide (ATO) only when accounting for these memory effects. Controlled prior subculture led to the finding of a synergistic combination treatment of ATO with the thioredoxin reductase 1 inhibitor auranofin, which may prove useful in the management of NRF2-mediated resistance mechanisms.

Chili pepper extracts, capsaicin, and dihydrocapsaicin as potential anticancer agents targeting topoisomerases.

DNA topoisomerases regulate conformational changes in DNA topology during normal cell growth, such as replication, transcription, recombination, and repair, and may be targeted for anticancer drugs. A DNA topology assay was used to investigate DNA-damaging/protective activities of extracts from Habanero Red (HR), Habanero Maya Red (HMR), Trinidad Moruga Scorpion (TMS), Jalapeno (J), Serrano pepper (SP), Habanero Red Savina (HRS), Bhut Jolokia (BJ), and Jamaica Rosso (JR) peppers, demonstrating their inhibitory effect on the relaxation of pBR by Topo I. DNA topoisomerase II (Topo II) is proven therapeutic target of anticancer drugs. Complete inhibition of Topo II was observed for samples TMS, HR, and HMR. Extracts J and SP had the lowest capsaicin and dihydrocapsaicin content compared to other peppers. HR, HMR, TMS, J, S, HRS, BJ, JR extracts showed the anticancer effect, examined by MTS and xCell assay on the in vitro culture of human colon carcinoma cell line HCT116.

Attenuating Colorectal Cancer Using Nine Cultivars of Australian Lupin Seeds: Apoptosis Induction Triggered by Mitochondrial Reactive Oxygen Species Generation and Caspases-3/7 Activation.

As Australian lupin cultivars are rich sources of polyphenols, dietary fibers, high-quality proteins, and abundant bioactive compounds with significant antioxidant, antidiabetic, and anticancer activities, this research work is aimed at investigating the colon cancer alleviation activity of nine cultivars of lupin seeds on HCT116 and HT29 colon carcinoma cell lines through anti-proliferation assay, measurement of apoptosis, and identification of the mechanism of apoptosis. Nine cultivars were pre-screened for anti-proliferation of HCT116 and HT29 cells along with consideration of the impact of heat processing on cancer cell viability. Mandelup and Jurien showed significant inhibition of HCT116 cells, whereas the highest inhibition of HT29 cell proliferation was attained by Jurien and Mandelup. Processing decreased the anti-proliferation activity drastically. Lupin cultivars Mandelup, Barlock, and Jurien (dose: 300 μg/mL) induced early and late apoptosis of colon cancer cells in Annexin V-FITC assay. The mechanism of apoptosis was explored, which involves boosting of caspases-3/7 activation and intracellular reactive oxygen species (ROS) generation in HCT116 cells (Mandelup and Barlock) and HT29 cells (Jurien and Mandelup). Thus, the findings showed that lupin cultivars arrest cell cycles by inducing apoptosis of colorectal carcinoma cells triggered by elevated ROS generation and caspases-3/7 activation.

Assessment of anticancer, antibacterial, antifungal, and antioxidant activities of Micractinium reisseri (Chlorophyta, Trebouxiophyceae) methanolic extract

This study looked at the anticancer, antimicrobial, and antioxidant properties of a methanolic extract of Micractinium reisseri. In vitro antitumor activity of M. reisseri methanolic extract revealed an inhibitory effect against MCF-7 and HCT-116 (breast carcinoma and colon carcinoma) cell lines. Antimicrobial activity of M. reisseri methanolic extract was estimated against Gram-negative bacteria (Pseudomonas aeruginosa RCMB 010,049 and Escherichia coli RCMB 010,052), Gram-positive bacteria (Streptococcus pneumonia RCMB 010,010 and Bacillus subtilis RCMB 010,067) and fungi (Aspergillus fumigatus RCMB 02,568 and Candida albicans RCMB 05,036). The extract showed promising results against all the tested microbes except C. albicans which gave negative results. The algal extract showed 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing and hydroxyl radical scavenging activity in a concentration-dependent manner with maximum scavenging activity at concentrations (0.001, 0.001, and 5 mg/ml) for each assay, respectively. High-performance liquid chromatography (HPLC) revealed that the active compounds are phenols, alkaloids, and flavonoids. In conclusion, M. reisseri methanolic extract exhibited effective anticancer, antimicrobial, and antioxidant activities.

Abstract 2384: Quantitative phase imaging provides new insights into diverse morphologies in cell lines and their behavior in response to alternative cancer treatments

Abstract Quantitative phase imaging (QPI) through holographic time-lapse microscopy allows for non-invasive measurements of cell dry mass and the ability to track cells for long periods of time with minimal damage, making it most beneficial for research on cancer treatment. Research with this enhanced method of observation can help decipher various morphologies present in specific cell lines and show how each cell can respond differently to provided signals. In this assay, we studied N2a murine neuroblast differentiation in response to Retinoic acid, as well as the anticancerous activity of Saffron and UV in MDA-MB-468 breast carcinoma and HCT116 colon carcinoma cell lines. Cells were observed using Telight’s Q-Phase, a multimodal microscope that utilizes QPI holographic time-lapse technology with label-free monitoring. The cells were able to survive in optimum conditions for up to 96 hours without any appreciative loss of viability. With the emergence of phytochemicals being used as alternative cancer treatments, specific pigments in saffron have shown therapeutic effects against cancer tissues such as apoptosis and decreased carcinogen presence. A microanalysis of the effects of saffron on reducing the invasion potential, viability, and tumorigenicity of breast cancer cell lines was performed. QPI microscopy was performed on MDA-MB-468 human breast adenocarcinoma and HCT116 colon carcinoma cell lines treated with varying doses of saffron and ultraviolet radiation. Cell lines were observed under the microscope for 58-hour periods for changes in mass, movement, and replication. The quantitative data showed a reduction of projections from saffron treatment but did not show any significant reduction in cell movement over the 58-hours. These results were later confirmed through scratch wound and Boyden chamber assays. Similarly, when examining the process of N2a neuroblast differentiation, it was observed that cells do not move towards differentiation in a linear manner, but rather changed their direction and path several times before committing to their final phenotype. These types of observations are critical in understanding the biological processes behind differentiation and drug response and could not have been done without using the various quantitative and analysis tools provided by the QPI technology. Citation Format: Rida Khan, Meaghan McDonald, Albina Mikhaylova, Fatima K. Rehman. Quantitative phase imaging provides new insights into diverse morphologies in cell lines and their behavior in response to alternative cancer treatments [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 2384.

Antitumor activity of triazine mimic antibiotics for DNA-binding implications (impressive activity in vitro against a variety of tumor types in the NCI-60 screen): NSC 710607 to fight HCT-116 human colon carcinoma cell lines in vivo using the hollow fiber assay and xenograft mouse models.

Successful clinical applications of DNA-directed selective cytotoxic agents disrupt the vital replication/transcription processes and ultimately lead to cancer cell death. This study aimed to examine the growth screen of two lead triazine compounds in a number of cell lines and xenografts and to develop anticancer agents with noncovalent binding affinity bringing fewer side effects. The NCI initial hollow fiber test was performed using an established procedure. The cytostatic and cytocidal capacities of the test compounds were assessed by evaluating cytotoxicity by simply performing a standard cellular viability assay. The nude mouse human tumor xenograft system was used as an in vivo model. More sensitive drug with sub-micromolar activity met the interdisciplinary criteria for testing and was referred to evaluations in subcutaneous colorectal carcinoma HCT-116 human tumor xenografted into nude mice. Principal findings of the study: total cytostasis, almost nontoxic schedules, specific working hypotheses, strong rationale for the potential use, and important general implications (relevance to human biology). NSC 710607 displayed in vivo better than Cisplatin and 5-fluorouracil abilities to significantly decrease tumor growth. Cell proliferation can be reduced or stopped in vivo in view of the xenograft results. The mimic molecule behaves as DNA-binding antitumor antibiotics with great potential as general anticancer agents and deserves further trials. NSC 710607 represents the result of a design strategy with outstanding potential. This study also identifies the prognostic significance and is likely to translate to other species or systems.

Comparative therapeutic effects of Pituranthos tortuosus aqueous extract and phyto-synthesized gold nanoparticles on Helicobacter pylori, diabetic and cancer proliferation

Synthesis of nanomaterials and nanoparticles represents a promising era in biological research that may lead to the development of new approaches in biomedicine and pharmaceutical studies. In this regard, the present study aimed to investigate anti-diabetic, anti-Helicobacter pylori and cytotoxic activities of phyto-synthesized gold nanoparticles using Pituranthos tortuosus (Desf.) aqueous extracts. Characterization of the phyto-synthesized gold nanoparticles (AuNPs) was performed using several analytical techniques including ultraviolet–visible spectroscopy, transmission electron microscopy, X-ray diffraction, Fourier transforms infrared spectroscopy, and Thermo gravimetric analysis. In addition, preliminary phytochemical screening of the P. tortuosus aqueous extract was tested using standard biochemical methods. Anti-diabetic activity was evaluated by α-glucosidase inhibitory assay, cytotoxic activity was evaluated using cell viability MTT assay, and H. pylori activity was detected using the minimum inhibitory concentration (MIC). The initial phytochemical analysis of P. tortuosus aqueous extract indicated the existence of the following compounds: flavonoids, carbohydrates, glycosides, tannins, coumarins, proteins, alkaloids, terpenoids and essential oils. The phyto-synthesized AuNPS effectively inhibit the α-glucosidase resulting in an IC50 value of 77, 44 μg/ml. AuNPS showed anti-H. pylori activity, especially against the multi-drug resistant strains with an MIC value of 15, 63 (μg/ml). AuNPs showed significant cytotoxic activity against hepatocellular carcinoma cell lines HepG2 and human colon carcinoma cell lines HCT-116 with IC50=23.60 μg/ml and IC50=6.27 μg/ml, respectively. This study indicated that the phyto-synthesis AuNPs from P. tortuosus aqueous extract enhanced the anti-diabetic, anti-H. pylori and cytotoxic activities.

Antiproliferative, antimicrobial and antioxidant properties of Cedrus libani and Pinus pinea wood oils and Juniperus excelsa berry oil

In this paper we report the analysis of essential oils from the woods of C. libani and P. pinea and from the berries of J. excelsa. Several differences were observed with respect to previous studies. In particular, the wood of C. libani was mainly characterized by sesquiterpenes with himachalenes isomers, (E)- and (Z)-α-atlantones and α-acorenol (7.0%) as most representative compounds; in P. pinea wood oil and J. excelsa berries oil were observed differences mainly among the minor components with different isomers of some specific compounds not previously recognized. The oils were assessed in selected bioactivity tests. C. libani wood oil resulted the most active in both the antioxidant and cytotoxic (against human A375 malignant melanoma, human MDA-MB 231 breast adenocarcinoma and human HCT116 colon carcinoma cell lines) tests; on the other hand in the antimicrobial assay it was observed a different susceptibility of different bacteria to the different oils. All the oils exerted a remarkable activity against the yeast Candida albicans. Taken together, these results support some of the traditional uses of these three plants in the Mediterranean area and give evidence of their potential on an industrial level.

New Platinum (II) Ternary Complexes of Formamidine and Pyrophosphate: Synthesis, Characterization and DFT Calculations and In vitro Cytotoxicity.

Platinum (II) and platinum (IV) of pyrophosphate complexes have been prepared and characterized to discover their potential as antitumor drugs. This study was conducted to prepare and characterize new ternary platinum (II) complexes with formamidine and pyrophosphate as an antitumor candidate. The complexes have been characterized by mass, infrared, UV-Vis. spectroscopy, elemental analysis, magnetic susceptibility, thermal analyses, and theoretical calculations. They have been tested for their cytotoxicity, which was carried out using the fastcolorimetric assay for cellular growth and survival against MCF-7 (breast cancer cell line), HCT- 116 (colon carcinoma cell line), and HepG-2 (hepatocellular cancer cell line). All complexes are diamagnetic, and the electronic spectral data displayed the bands due to square planar Pt(II) complexes. The optimized complexes structures (1-4) indicated a distorted square planar geometry where O-Pt-O and N-Pt-N bond angles were 82.04°-96.44°, respectively. Results also show that all complexes are neutral, stable and non-hygroscopic and have noticeable cytotoxicity with IC50 (μM): 0.035-0.144 MCF-7(breast cancer cell line), 0.042-0.187 HCT-116 (colon carcinoma cell line), and 0.063-0.168 HepG-2 (hepatocellular cancer cell line). Moreover, the results show that the complex (4) has the best IC50 value. The complexes showed noticeable cytotoxicity and are considered as promising antitumor candidates for further applications.

Combined Targeted Proteomics and Oxylipin Metabolomics for Monitoring of the COX-2 Pathway.

The important role of inducible cyclooxygenase-2 (COX-2) in several diseases necessitates analytical tools enabling thorough understanding of its modulation. Analysis of a comprehensive oxylipin pattern provides detailed information about changes in enzyme activities. In order to simultaneously monitor gene expression levels, a targeted proteomics method for human COX-2 is developed. With limits of detection and quantification down to 0.25 and 0.5 fmol (on column) the method enables sensitive quantitative analysis via LC-MS/MS within a linear range up to 2.5pmol. Three housekeeping proteins are included in the method for data normalization. A tiered approach for method development comprised of in silico and experimental steps is described for choosing unique peptides and selective and sensitive SRM transitions while avoiding isobaric interferences. This method combined with a well-established targeted oxylipin metabolomics method allows to investigate the role of COX-2 in the human colon carcinoma cell lines HCT-116, HT-29, and HCA-7. Moreover, the developed methodology is used to demonstrate the time-dependent prostanoid formation and COX-2 enzyme synthesis in lipopolysaccharide-stimulated human primary macrophages. The described approach is a helpful tool which will be further used as standard operation procedure, ultimately aiming at comprehensive targeted proteomics/oxylipin metabolomics strategies to examine the entire arachidonic acid cascade.

Cytotoxic and chemotaxonomic study of isolated metabolites from Centaurea aegyptiaca

AbstractThe aerial parts of Centaureaa egyptiaca afforded 10 secondary metabolites including four sesquiterpene lactones; chlorohyssopifolin A (centaurepensin) (1), rediolpidetriol (2), linichlorinA (3), and sinaicin (4), one monoterpene; loliolid (5), one phenolic: tyrosol (6), three lignans; arctigenin (7), matairesinol (8), and pinoresinol (9), and one steroid; ergosta‐5,22‐dien‐3‐ol (10). The cluster analysis of 32 Centaurea species revealed that C. aegyptiaca is closely related to C. repens and C. solstitialise. The isolated compounds (1–10) were screened against CCRF‐CEM‐leukemia, MDA‐MB‐231‐pcDNA3 breast cancer, and HCT116 (p53+/+) colon carcinoma cell lines. Compounds1 and 2 were the most potent compounds against both leukemia and breast carcinoma cell lines.

Antitumor, Antimicrobial activities and Phytochemicals Constituent of different Extracts of Pulicaria undulata (Forssk.) Oliver. Grown Naturally in Saudi Arabia

Antitumor and antimicrobial resistance are a habitual global issue, which continually demands finding new natural compounds to encounter the resistance. Pulicaria undulata (Forssk.) Oliver. (Asteraceae family) has numerous promising medicinal properties. The recent work aimed at determination of antitumor effects of three extracts of P. undulata on three types of human carcinoma; HEPG-2 hepatocellular carcinoma, MCF-7 breast carcinoma and HCT-116 colon carcinoma cell lines. Anticancer activity was assessed through studying the viability of the cancer cells and apoptotic pathway. Also, antimicrobial potency of different extracts was assessed against studied human pathogens (five Gram negative bacteria, two Gram positive bacteria and yeast). The results reveal that chloroform extract has different levels of cytotoxicity toward the three types of cancer cell lines. A considerable decline in cancer cell rates has been linked to increasing in concentration of plant extract. The half maximal inhibitory concentration IC 50 value was 3.01 µg/ mL for the HepG-2, 16.4 µg/mL for the MCF-7, and 7.4 µg/ mL for HCT-116. Followed by the ethyl acetate extract which showed strong cytotoxic activity against HEPG2 with IC 50 = 12.2 µg ∕ml and moderate activities against MCF7 and HCT 116 and recorded (IC 50 = 26.7 and 26.4 µg ∕ml, respectively). While the crude methanol extract recorded the lowest cytotoxic effect against HEPG2, MCF7 and HCT 116 with (IC 50 = 51.4, 105.1 and 86.7 µg ∕ ml, respectively). Chloroform and ethyl acetate extracts have a high antimicrobial activity more than methanol extract against the pathogens being studied. HPLC and GCMs Analysis identified numerous chemical compounds of P. undulata extracts with various therapeutic benefits. In conclusion, P. undulata has the potential to act as an antimicrobial agent against various pathogenic microbes and is a promising wild herb for the treatment of cancer.

SYNTHESIS AND ANTIPROLIFERATIVE ACTIVITY OF SALICYLIDENEHYDRAZONES BASED ON INDOLE-2(3)-CARBOXYLIC ACIDS

A series of indole-2(3)-carboxylic acid hydrazones were synthesized and their antiproliferative activity was studied for the K562 leukemia, MCF7 breast adenocarcinoma, and HCT116 colon carcinoma cell lines and resistant HCT116p53KO subline in order to preliminary screen biological properties and to reveal the role of the aldehyde fragment and heterocycle in the ability of compounds to inhibit tumor growth. A number of derivatives have been identified that inhibit the growth of tumor cells in submicromolar concentrations, promising for further optimization and in-depth study of antitumor properties.

Synthesis and antiproliferative activity of salicylidenehydrazones based on indole-2(3)-carboxylic acids

A series of indole-2(3)-carboxylic acid hydrazones were synthesized and their antiproliferative activity was studied for the K562 leukemia, MCF7 breast adenocarcinoma, and HCT116 colon carcinoma cell lines and resistant HCT116p53KO subline in order to preliminary screen biological properties and to reveal the role of the aldehyde fragment and heterocycle in the ability of compounds to inhibit tumor growth. A number of derivatives have been identified that inhibit the growth of tumor cells in submicromolar concentrations, promising for further optimization and in-depth study of antitumor properties.

Design, Synthesis and In Vitro Cytotoxic Activity of New 6,9-Disubstituted Purine Analogues

A series of new 6,9-disubstituted purine analogs with 4-substituted piperazine at C-6 and 4-substituted benzyl at N-9 were designed and synthesized in four steps. All synthesized compounds (7-26) were screened initially for their in vitro anticancer activity on Huh7 liver, HCT116 colon and MCF7 breast carcinoma cell lines. Cytotoxic bioactivity studies revealed that all compounds screened, with compound 19 being the exception, were found to have promising cytotoxic activities at IC50 range of 0.05-21.8 ∆M against cancer cells Huh7, HCT116 and MCF7. Among the prepared purine analogs, two of them (12 and 22) exhibited excellent cytotoxic activities, with IC50 0.08-0.13 ∆M, on Huh7 cells comparable to camptothecin (CPT) and better than cladribine, fludarabine and 5-FU. Afterwards, the evaluation of cytotoxicity of the most potent purine analogs was screened against further hepatocellular cancer (HCC) cell lines. The 6-(4-(4-trifluoromethylphenyl)piperazine (12) and 6-(4-(3,4-dichlorophenyl)piperazine analogs (25) displayed a significant IC50 values (IC50 < 0.1-0.13 ∆M) comparable to CPT and better cytotoxic bioactivity when compared with 5-FU, cladribine and fludarabine on HCC cells (Huh7 and HepG2).

Single Spheroid Metabolomics: Optimizing Sample Preparation of Three-Dimensional Multicellular Tumor Spheroids.

Tumor spheroids are important model systems due to the capability of capturing in vivo tumor complexity. In this work, the experimental design of metabolomics workflows using three-dimensional multicellular tumor spheroid (3D MTS) models is addressed. Non-scaffold based cultures of the HCT116 colon carcinoma cell line delivered highly reproducible MTSs with regard to size and other key parameters (such as protein content and fraction of viable cells) as a prerequisite. Carefully optimizing the multiple steps of sample preparation, the developed procedure enabled us to probe the metabolome of single MTSs (diameter range 790 ± 22 µm) in a highly repeatable manner at a considerable throughput. The final protocol consisted of rapid washing of the spheroids on the cultivation plate, followed by cold methanol extraction. 13C enriched internal standards, added upon extraction, were key to obtaining the excellent analytical figures of merit. Targeted metabolomics provided absolute concentrations with average biological repeatabilities of <20% probing MTSs individually. In a proof of principle study, MTSs were exposed to two metal-based anticancer drugs, oxaliplatin and the investigational anticancer drug KP1339 (sodium trans-[tetrachloridobis(1H-indazole)ruthenate(III)]), which exhibit distinctly different modes of action. This difference could be recapitulated in individual metabolic shifts observed from replicate single MTSs. Therefore, biological variation among single spheroids can be assessed using the presented analytical strategy, applicable for in-depth anticancer drug metabolite profiling.

Low-dimensional compounds containing bioactive ligands. Part XI: Synthesis, structures, spectra, in vitro anti-tumor and antimicrobial activities of 3d metal complexes with 8-hydroxyquinoline-5-sulfonic acid

Five 3d metal [M(H2O)2(SQ)]n complexes, where M is Mn (1), Co (2), Ni (3), Zn (4) and Cu (5), and SQ is the dianion of 8-hydroxyquinoline-5-sulfonic acid have been prepared and characterized by IR spectroscopy and elemental analysis. X-ray structure analysis of 1 and 5 in combination with IR spectroscopy of all five complexes showed that complexes 1–4 are isostructural 2D polymers, while 5 is 1D polymer due to the different bonding character of SQ. Stability of all complexes in DMSO was proved by UV–Vis spectroscopy and for 4 also verified by 1H and 13C NMR spectroscopy. Cytotoxic activity tests against cervical carcinoma HeLa, colon carcinoma HCT116 and normal human fibroblast MRC-5 cell lines showed moderate but selective cytotoxicity of all complexes. Complex 1 exhibited highest activity with IC50 = 39.1 and 33.4 µM, respectively, but is lower than activity of cisplatin (CP) which IC50 = 24.3 and 14.3 µM, respectively. However, their cytotoxic activity against healthy MRC-5 cells (IC50 > 92 µM) is much lower than CP (IC50 = 19.7 µM). Antimicrobial activity of all complexes against 9 strains of pathogenic bacteria, five mould species and two yeast species is negligible.

Lamellarin Sulfates from the Pacific Tunicate Didemnum ternerratum.

Six new lamellarin sulfates (1-6) were isolated from the methanolic extract of the Pacific tunicate Didemnum ternerratum, collected from the Kingdom of Tonga. Mass spectrometric molecular networking through the GNPS platform was used to target the isolation of 1-6. Planar structures were elucidated through a combination of NMR and MS experiments. Through comparison of experimental and calculated ECD spectra, the absolute configurations of atropisomers 2-5 were determined, with their energetic barriers to racemization also determined computationally. The cytotoxicity of the compounds was tested against the human colon carcinoma cell line HCT-116, where lamellarin D-8-sulfate (5) exhibited moderate activity with an IC50 of 9.7 μM.

Abstract 4851: Involvement of extracellular vesicles in the abscopal effect induced by oncolytic adenovirotherapy

Abstract Background: The abscopal effect is a phenomenon that shrinkage of metastatic tumors occurs concurrently with shrinkage of primary tumors by local treatments such as radiotherapy, and is generally considered to be caused through activation of the immune system. Extracellular vesicles (EVs) are recent interesting topics in scientific fields and are known to play important roles in a variety of intercellular communication processes including immune response. We previously developed a telomerase specific oncolytic adenovirus, Telomelysin (OBP-301), which is currently being tested in clinical trials, and have sometimes observed therapeutic effects on metastatic tumors after local injection to primary tumors in immunodeficient mouse models. Here, we hypothesized that EVs derived from OBP-301-treated tumors may be associated with the abscopal effect of OBP-301. Methods: A human colon carcinoma cell line HCT116 and HCT116 expressing RFP (HCT116-RFP) were used in this study. EVs were isolated by ultracentrifugation from HCT116 cells treated with OBP-301 (Exo-301). Intercellular communication and cytotoxic activity via EVs after OBP-301 treatment were assessed in a non-contact co-culture system in vitro and a bilateral subcutaneous tumor mouse model in vivo. Results: Exo-301 showed potent cytotoxic effects on HCT116 cells and this cytotoxicity was significantly blocked in the presence of anti-CD9 antibody, which suggested that Exo-301, not OBP-301 itself, was involved in this cytotoxicity. In non-contact co-culture experiments, more RFP was taken up into HCT116 cells after OBP-301 treatment on HCT116-RFP cells than control treatments, which meant that EVs including RFP reached HCT116 cells through membrane after OBP-301 treatment on HCT116-RFP cells. In a HCT116 bilateral subcutaneous tumor model in BALB/c nude mice, intratumoral injection of OBP-301 significantly suppressed the growth of untreated tumors as well as treated tumors, and in a bilateral tumor model with HCT116 on one side and HCT116-RFP on the opposite side, immunohistochemical staining showed the presence of RFP in untreated HCT116 tumors after OBP-301 treatment on HCT116-RFP tumors. These findings suggested that EVs derived from OBP-301-treated tumors was possibly involved in some cytotoxic effects on the opposite untreated tumors. Conclusion: EVs may be an important factor in the abscopal effect brought by local treatment of OBP-301 besides immune activation. Citation Format: Yoshihiko Kakiuchi, Shinji Kuroda, Nobuhiko Kanaya, Kento Kumon, Tomoko Tsumura, Satoru Kikuchi, Masahiko Nishizaki, Shunsuke Kagawa, Hiroshi Tazawa, Yasuo Urata, Toshiyoshi Fujiwara. Involvement of extracellular vesicles in the abscopal effect induced by oncolytic adenovirotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4851.