Colon Cancer Cell Line HT-29 Research Articles

Phytoconstituents, Antioxidant Activity and Cytotoxicity of Puya chilensis Mol. Extracts in Colon Cell Lines

Puya chilensis Mol. is a plant of the Bromeliaceae family, which has been traditionally used for medicinal applications in various digestive disorders. In this study, the phytoconstituents of six extracts of stems and flowers of P. chilensis were evaluated: phenols, flavonoids and total anthraquinones, as well as their antioxidant capacity and cytotoxicity in colon cancer cell lines HT-29. The data demonstrate that the ethyl acetate extract of P. chilensis flowers is cytotoxic in HT-29 cell lines (IC50 = 41.70 µg/mL) without causing toxic effects on healthy colon cells (IC50 > 100 µg/mL); also, this extract concentrated the highest amount of phenols (4.63 μg GAE/g d.e.), flavonoids (31.5 μg QE/g d.e.) and anthraquinones (12.60 μg EE/g d.e.) among all the extracts tested, which also correlated with its highlighted antioxidant capacity (DPPH∙IC50 = 4.15 mg/mL and FRAP 26.52 mM TEAC) over the other extracts. About thirty-five compounds were identified in this extract−the fatty acid esters present have been shown to have therapeutic effects on several types of cancer and could explain its antiproliferative activity.

Exploring the impact of sEH inhibition on intestinal cell differentiation and Colon Cancer: Insights from TPPU treatment

Inhibition of soluble epoxide hydrolase (sEH) appears to be promising for the treatment of many diseases. Studies have focused on the beneficial effects of epoxyeicosatrienoic acids (EETs), which are sEH substrates. However, our recent studies have shown that the sEH activity is crucial for the proper intestinal cell differentiation. In this recent study, we investigated the impact of TPPU, an inhibitor of sEH, on the colon cancer cell lines Caco2 and HT-29. We analysed the changes in the expression of the cytoskeletal protein ezrin and the phosphorylated protein kinase p38 (p-p38). Our results showed a decrease in ezrin expression in differentiated cells and an increase in p-p38 expression after TPPU treatment. Immunocytochemical staining revealed a higher staining intensity of p-p38 in the nuclei of HT-29 cells following TPPU treatment. Immunohistochemical staining was performed on human samples of normal colon tissue, grade 2 tumours, and embryonal/foetal tissues. The staining intensity of ezrin in tumours was reduced in the surface area compared to the crypts. Additionally, we observed the translocation of p-p38 expression from the cytoplasm to the nucleus during differentiation. The tumour samples exhibited higher levels of p-p38 in the cytoplasm, similar to normal undifferentiated tissue. To observe the disruption of the cytoskeleton after TPPU treatment, confocal microscopy was used. It was found that β-actin associated with ezrin forms clusters under the plasma membranes. All of these results are significant because sEH inhibitors are being tested in clinical trials, but they could cause an unexpected adverse effects.

Fucosylated Chondroitin Sulfate from Bohadschia ocellata: Structure Analysis and Bioactivities

Fucosylated chondroitin sulfate (FCS) was prepared from Bohadschia ocellata using protease hydrolysis. The structural characteristics of FCS were confirmed through chemical composition analysis using FTIR spectroscopy, 1H NMR, and 13C NMR. FCS from B. ocellata (FCS-Bo) exhibited an average molecular weight of approximately 122 kDa. The biological activities of FCS-Bo, including anticoagulant, anti-cancer, and Protein Tyrosine Phosphatase 1B (PTP1B) inhibition, were evaluated. FCS-Bo displayed potent anticoagulant properties, markedly extending activated partial thromboplastin time, prothrombin time, and thrombin time when compared to the heparin control. In anti-cancer bioactivity research, FCS-Bo efficiently inhibited colony formation in the colon cancer cell lines HCT-116, HT-29, and DLD-1, achieving inhibition rates of up to 65%. Additionally, FCS-Bo exhibited significant inhibition of PTP1B, with an IC50 as low as 0.0326 µg/mL, suggesting its potential for improving insulin sensitivity and managing conditions such as type 2 diabetes and obesity.

Platinum(II) acetimino cyclometalated complexes derived from room temperature ammonia activation

Two Pt(II) rollover cyclometalated complexes, [Pt(bpyPh‐H)(DMSO)Me] (1) and [Pt(bpyPh‐H)(DMSO)Cl] (2), where bpyPh‐H is the C3‐deprotonated 6‐phenyl‐2,2’‐bipyridine, were reacted with aqueous ammonia in acetone at room temperature. In the case of the electron‐poor complex 2 simple substitution of DMSO gave the amino complex [Pt(bpyPh‐H)(NH3)Cl] (4), whereas the electron‐rich complex 1 produced the rare acetimine complex [Pt(bpyPh‐H)(HN=CMe2)Me] (3), stable both in solution and in the solid state. The result is noteworthy, as the condensation product of acetone and ammonia, acetimine, is not stable under mild conditions. The reaction likely requires a Pt(II) electron‐rich complex and the presence of a carbon donor in trans position. An analogous reaction occurs with methylamine, allowing the synthesis of the secondary imine complex [Pt(bpyPh‐H)(MeN=CMe2)Me], 5. Starting from this finding a series of cyclometalated acetimine complexes [Pt(N^C)(HN=CMe2)Me] were isolated and characterized with other classical and rollover cyclometalated ligands with an array of different electronic and steric properties, indicating a possible extension of the reaction to various cyclometalated ligands. A preliminary study on the antimicrobial and antitumor properties of complex 3, taken as a model for this class, showed no significant effects against Gram‐positive, Gram‐negative bacteria, or yeasts, but found activity in vitro against HT29 colon cancer cell line.

Design, Synthesis, Biological Evaluation and Molecular Modeling Studies of New Aminopyrimidine Derivatives as Potential Anticancer Agents

A wide range of bioactive compounds have been created using the versatile pharmacophore pyrimidine. In order to improve biological applications, we, therefore, envisioned adding these privileged moieties. In this work, we developed an efficient 2-aminopyrimidine derivatives. We further investigated the anticancer activity of all the compounds against colon cancer cell lines COLO-205 and HT-29. Out of several compounds synthesized, two compounds SD2 and SD4 have shown comparable potent anticancer activity when compared to standard Cabozantinib. The most potent cytotoxic compound against colon cancer cells in our study was found to be SD2 with an IC50 value of 9.57 [Formula: see text]M. The investigation from molecular docking showed the best binding affinity. The complex stability within an enzyme’s pocket was demonstrated by the molecular dynamic simulation running for 100 ns.

Systematic bioinformatics analysis reveals the role of shikonin in blocking colon cancer progression by identifying senescence-induced genes.

Shikonin, a naturally occurring naphthoquinone compound extracted from comfrey plants, has antitumor, anti-inflammatory, and antimicrobial properties. Cell senescence plays a key role in preventing tumor progression. It is unclear whether shikonin has an effect on cell senescence in colon cancer. In the current study, we first determine the IC50 values of shikonin on colon cancer cell lines HT29 and HCT116. Then, we verified the inhibitory effects of shikonin on the proliferation and migration abilities of colon cancer cell lines HT29 and HCT116 using cell counting kit-8, colony formation, and wound healing assays. Next, we identified a series of potential targets using high-throughput mRNA sequencing and identified 210 upregulated and 296 downregulated genes. KEGG profiling revealed eight downregulated genes associated with cell senescence: CCNB3, IL-1α, CXCL8, CDKN2A, MYC, IGFBP3, SQSTM1, and GADD45G. Among them, CXCL8 and CDKN2A were associated with poor prognosis in patients with colon cancer, suggesting that their downregulation by shikonin could improve patient survival. Furthermore, SA-β-galactosidase staining revealed that the percentage of cellular senescence in colon cancer cells was significantly increased after shikonin treatment. Molecular docking revealed that shikonin suppressed colon cancer progression by blocking CXCL8 activity. Based on these findings, we deem that shikonin might induce senescence and exert antitumor activity in colon cancer cells by downregulating CDKN2A and CXCL8. This provides a new molecular mechanism and potential therapeutic target for shikonin to inhibit colon cancer progression.

Hsa-miR-181a-2-3p inhibits the oncogenicity of colon cancer by directly targeting STING.

Colon cancer is a common malignant tumor of the gastrointestinal system, which is characterized by high morbidity and mortality. The purpose of this study was to analyze the expression and biological role of miR-181a-2-3p in colon cancer and to investigate the molecular mechanism of its regulatory effect on colon cancer through stimulator of interferon genes (STING). Real-time reverse transcription polymerase chain reaction (qRT-PCR) assay was used to detect the expression of miR-181a-2-3p in colon cancer cell lines and normal intestinal epithelial cells. After overexpression of miR-181a-2-3p in colon cancer cell lines SW480 and HT29, cells were examined by CCK8, Transwell, and flow cytometry assays for alterations in proliferation, migration, apoptosis, and cell cycle. Target genes of miR-181a-2-3p were predicted by bioinformatics and validated by dual luciferase assays. Rescue experiments were performed to explore the role of STING in the effect of miR-181a-2-3p. The effect of miR-181a-2-3p on colon cancer proliferation in vivo was validated by nude mouse tumorigenicity assay. miR-181a-2-3p was lowly expressed in both colon cancer tissues and cell lines. Overexpression of miR-181a-2-3p led to reduced proliferation and migration, increased apoptosis, and altered cell cycle in colon cancer cell lines SW480 and HT29. STING was a target gene of miR-181a-2-3p. Increased STING expression partially counteracted the effect of overexpression of miR-181a-2-3p on colon cancer cell lines. miR-181a-2-3p also suppressed colon cancer proliferation in vivo. miR-181a-2-3p inhibits the proliferation and oncogenicity of colon cancer, and its molecular mechanism could be inhibited by STING.

Decoration of silver nanoparticles over chitosan-Arabic gum hydrogel: Synthesis, characterization and investigation of its antioxidant and anti-colorectal cancer activities

The present study describes the production of hybrid biopolymer hydrogel supported Ag NPs in a sustainable manner, along with its biological investigations and characterizations. A composite hydrogel of chitosan and Arabic gum (CS-AG) was utilized as the support or template for the green synthesis of nanoparticles. By taking advantage of their electron-rich organo-functions, the biopolymers could function as both an encapsulating stabilizer of the Ag NPs and a green-metric reductant of the incoming Ag ions. The physicochemical properties of the material were evaluated using an array of advanced analytical techniques like X-ray diffraction (XRD), elemental mapping, scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDX), Fourier Transformed Infrared Spectroscopy (FT-IR), UV–Vis spectroscopy and Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). Towards the biological implication, the material was used in anticancer studies against HT-29 and Caco-2 colon cancer cell lines using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)colorimetric method, as well as in the biological evaluation of antioxidant properties by the DPPH (2,2-diphenyl-1-picrylhydrazyl)method. The substance appeared to have strong anticancer properties based on the DPPH assay's noteworthy IC50 value. Remarkably, it was discovered that the percentage of malignant cell lines' cell viability decreased over the Ag NPs@CS-AG nanocomposite in a dose-dependent manner. The MTT assay produced the corresponding IC50 values against the two cell lines, which were 366.1 μg/mL and 264.5 μg/mL, respectively. At 545 nm, the absorbance rate was assessed, showing toxicity on normal cell line (CHO) up to 1927 μg/mL, which was quite insignificant.

Effects of photobiomodulation on colon cancer cell line HT29 according to mitochondria

Background/AimAlthough photobiomodulation therapy (PBMt) is available to alleviate post-operative side effects of malignant diseases, its application is still controversial due to some potential of cancer recurrence and occurrence of a secondary malignancy. We investigated effect of PBMt on mitochondrial function in HT29 colon cancer cells. MethodsHT29 cell proliferation was determined with MTT assay after PBMt. Immunofluorescent staining was performed to determine mitochondrial biogenesis and reactive oxygen species (ROS). Mitochondrial membrane potential was measured with Mitotracker. Western blotting was executed to determine expression of fission, fusion, UCP2, and cyclin B1 and D1 proteins. In vivo study was performed by subcutaneously inoculating cancer cells into nude mice and immunohistochemistry was done to determine expression of FIS1, MFN2, UCP2, and p-AKT. ResultsThe proliferation and migration of HT29 cells reached maximum with PBMt (670 nm, light emitting diode, LED) at 2.0 J/cm2 compared to control (P < 0.05) with more expression of cyclin B1 and cyclin D1 (P < 0.05). Immunofluorescent staining showed that ROS and mitochondrial membrane potential were enhanced after PBMt compared to control. ATP synthesis of mitochondria was also higher in the PBMt group than in the control (P < 0.05). Expression levels of fission and fusion proteins were significantly increased in the PBMt group than in the control (P < 0.05). Electron microscopy revealed that the percentage of mitochondria showing fission was not significantly different between the two groups. Oncometabolites including D-2-hydoxyglutamate in the supernatant of cell culture were higher in the PBMt group than in the control with increased UCP2 expression (P < 0.05). Both tumor size and weight of xenograft in nude mice model were bigger and heavier in the PBMt group than in the control (P < 0.05). Immunohistologically, mitochondrial biogenesis proteins UCP2 and p-AKT in xenograft of nude mice were expressed more in the PBMt group than in the control (P < 0.05). ConclusionsTreatment with PBM using red light LED may induce proliferation and progression of HT29 cancer cells by increasing mitochondrial activity and fission.

Quantum sized blue light emitting ZnO nanoparticles: Synthesis, characterization and biomedical applications

The synthesis of zinc oxide nanoparticles via the hydrolysis and condensation method is described in this study. Characterization of the synthesized nanoparticles through X-ray diffraction, optical absorption, photoluminescence, and transmission electron microscopy discloses highly stable particles with an exceptionally small and uniform size distribution, showing a discrete quantum size effect. These nanoparticles show robust blue emission and demonstrate potent antibacterial activity against a spectrum of pathogenic bacteria, making them capable candidates for combating infectious diseases. Additionally, they display noteworthy anti-cancer properties against HT-29 colon cancer cell lines. This dual functionality of the ZnO nanoparticles holds significant promise for the development of affordable therapies in biomedical applications, as well as for potential applications in electronics.

Two Metallic Nanoparticles Formulas of Phyllostachys heterocycla Extract Exhibited Potent Cytotoxicity against Ovarian Cancer Cells through Apoptosis Induction

Phyllostachys heterocycla is well-known for its high diversity of bioactive metabolites, which are the reason for its various potential medical uses for which anticancer activity has been proven. Herein, Phyllostachys heterocycla extract was prepared in two different metallic nanoparticle formulas such as iron oxide nanoparticle-boron, and iron oxide nanoparticle-humic acid (Fe2O3 NP-B and Fe2O3 NP-HA) with average particle sizes of 12.25 nm and 15.80 nm, respectively. Phyllostachys heterocycla extract and the two nano-formulas were investigated to obtain their cytotoxic activity. The crude extract exhibited potent cytotoxic activity against the ovarian (OVCAR-3) cancer cell line, with IC50 values of 16.3 µg/mL. In comparison, the two nano-loaded forms displayed a much more promising cytotoxic activity with IC50 values of 0.9 µg/mL for Fe2O3 NP-HA, and 6.4 µg/mL for Fe2O3 NP-B. Additionally, NP-HA and NP-B showed potent cytotoxic activities against prostate (PC-3) and pancreatic (Panc1) cancer cell lines with IC50 values of 2.31, 6.3 µg/mL for Fe2O3 NP-HA, and 14.9, 16.8 µg/mL for Fe2O3 NP-B. For apoptosis investigation, Fe2O3 NP-HA induced total ovarian apoptotic cell death by a 87.34-fold change, and necrosis by 1.29-fold change. Regarding cell cycle analysis, Fe2O3 NP-HA-PHE arrested the cell proliferation of OVCAR-3 cells in S-phase, with an increased cell population at S-phase of 42.6%. Additionally, it confirmed the apoptosis mechanism by inhibiting the antiapoptotic gene and activating the proapoptotic gene markers. Moreover, upon continuation of our phytochemical investigation of the plant, additional chemical components of the crude extract of Phyllostachys heterocycla were isolated using various chromatographic techniques. As a result, six compounds were isolated. By using different spectroscopic data, the chemical structures of the pure isolated compounds were assigned as stigmasterol (1), glyceryl monobehenate (2), vanillic acid (3), ferulic acid (4), catechin (5), and thymidine (6). These isolated compounds were previously reported for their potent cytotoxic activities against panel of cancer cell lines including pancreatic cancer and prostate cancer cell lines (Ferulic acid), beside the anti-tumor potential against ovarian cell lines (Stigmasterol). In addition to the cytotoxic activity against human larynx carcinoma HepG-2 cell lines (Catechin), human breast cancer MCF7 (Thymidine), and human colon cancer cell line HT-29 (Vanillic acid). Which may explain the significant cytotoxic and anticancer properities of the crude extract of Phyllostachysheterocycla.

Novel 2-pyridone Analogs with Anti-cancer Activity: Synthesis, In silico, and In vitro Evaluation

Background: 2-pyridone is frequently used to synthesize and develop new bioactive molecules approved for treating many diseases. The produced compounds play a significant role in inhibiting cancer growth. Objectives: Through a molecular docking investigation, we determined the binding affinity of 2-pyridone compounds with the Matrix Metalloproteinase receptor, which allowed us to develop, produce, and test the in vitro anticancer efficacy of those compounds. Method: 2-pyridones (A1-A12) were synthesized in a multistep process, followed by spectrum analysis to confirm the structure. In silico screening of the synthesized compounds was carried out with the assistance of AutoDock software. Flow cytometry was used on the HT-29 colon cancer cell line to measure A1-A12’s anticancer effect in a lab setting. Results: The enzyme matrix metalloproteinase receptor and A1-A12 interacted unexpectedly during a docking study (MMP3, MMP9 &amp; MMP13). Research has shown a strong affinity for MMP3 receptors for A9, A10, A11, A12, and A4, respectively. Further flow cytometric testing revealed compound A9 (R1) to be highly cytotoxic, with an IC50 value of 20.77 M. The anticancer activity of A9 (R1) against HT-29 colon cancer cell lines was also confirmed by in vitro results. Conclusion: These findings suggested that 2-pyridone compounds have promising therapeutic potential for cancer treatment, and more research on these lead moieties would be advantageous to discovering an effective anticancer drug.

Assessment of the Anticancer, Antimicrobial, and Antioxidant Activities of the Peganum harmala L. Plant

Introduction Synthetic chemical drug treatments have drawbacks including adverse effects and toxicity. Natural alternatives are sought after. Chemotherapy can be toxic. Medicinal plants offer a suggested alternative. This study examines P. harmala plant extracts for phenolic and flavonoid content, antimicrobial, antioxidant, and anticancer activities. Methods Two samples of P. harmala plant extract were prepared using ultrasonication, with ethanol concentrations of 100% and 50%. The total phenolic and flavonoid contents were determined using a chemical assay method. The antimicrobial activity of the extracts was evaluated against gram-positive bacteria ( Staphylococcus aureus and Streptococcus) and gram-negative bacteria ( Escherichia coli) using the diffusion procedure. The antioxidant impact of the extracts was assessed using the DPPH procedure. P. harmala extracts showed inhibitory effects on MCF7 breast cancer and HT29 colon cancer cell lines. Results The results of the study indicated that the P. harmala plant extracts were rich in phenolic compounds (with total phenolic content of 215.8 ± 3.5 and 155.8 ± 2.9 mg gallic acid per g extract for 50% and 100% ethanol extracts, respectively) and flavonoids (with total flavonoids content of 112.1 ± 3.1 and 92.3 ± 1.8 mg catechin per g extract) and had a high rate of antioxidant activity. The 50% ethanol extract yielded 411.8 ± 3.5 μmol trolox/g, while the 100% ethanol extract yielded 312.9 ± 8.2 μmol trolox per g extract. P. harmala extracts exhibited potent antimicrobial activity against E. coli, S. aureus, and Streptococcus. They also demonstrated strong anticancer activity, causing significant cell death in breast and colon cancer cell lines within 48 h of culturing. Conclusion P. harmala ethanolic extracts are rich in polyphenolic compounds and flavonoids, displaying high antioxidant activity. They also exhibit strong inhibitory effects against gram-positive and gram-negative bacteria and demonstrate potent anticancer activity against breast and colon cancer cell lines.

Cytotoxic and apoptotic effects of Prunus spinosa fruit extract on HT-29 colon cancer line

Colon cancer holds the position of the third most common type of cancer and stands as the third leading cause of cancer-related deaths for both men and women. Modern strategies in cancer prevention center around the use of natural compounds, which demonstrate a range of effects, including preventive, inhibitory, and latency-inducing impacts on the progression of cancer. In the present study, aqueous extracts derived from the fruits of Prunus spinosa L. (blackthorn, Rosaceae) are employed to assess their cytotoxic potential against the HT-29 colon cancer cell line. The fruit extract is administered to the HT29 cell line in different concentrations over 24 and 48-hours to evaluate the induction of apoptosis. The MTT cell viability test is employed to quantify the cytotoxic effect, indicating the extent of the impact. Additionally, the EB/AO (ethidium bromide/acridine orange) dual staining method is utilized to gather supplementary information regarding the cytotoxic effects. Observations after 24 hours of exposure showed no significant cytotoxic effect; however, 48-hour exposure revealed IC20, IC50, and IC80 values of 1.27, 173.7, and &gt; 1000 µg/ml, respectively, as determined by MTT analysis. Correspondingly, values of 5.06, 123.8, and &gt; 1000 µg/ml were recorded by the EB/AO dual staining method. Our results show that P. spinosa fruit water extract has an inhibitory effect on the HT-29 cell viability by exerting cytotoxic and apoptotic effects in a concentration-dependent and time-dependent manner. Toxicity studies have shown that MTT and EB/AO support each other and achieve similar results. Further extensive research into the metabolic and functional effects of P. spinosa could illuminate its potential and increase its economic importance in the field of anticancer treatments as a natural drug.

Spectroscopic analysis, quantum computational, and molecular docking investigations on the crystalline salt of 4-phenyl sulfanyl butanoic acid and piperazine with enhanced cytotoxic activity

In this investigation, the target crystalline salt was synthesized by combining 4-phenyl sulfanyl butanoic acid (PSBA) and piperazine (PZ). The process encompassed experimental methodologies, such as Powder X-ray Diffraction (PXRD),Single Crystal X-ray Diffraction (SCXRD), Nuclear Magnetic Resonance (NMR), Infrared Spectroscopy (IR),Thermogravimetric Analysis and Differential Scanning Colorimetry (TGA-DSC), as well as theoretical analyses of the synthesized compound.The crystalline salt was successfully synthesized via the solvent evaporation method at room temperature.PZ-PSBA has an orthorhombic crystal system with space group Pbca and unit cell parameters a = 7.4357(10)Å, b = 9.4853(12)Å, c = 34.299(5), and α=β=γ=90°. We employed the B3LYP functional with 6–311++G (d, p) as a basis set to geometrically optimize the molecular structure using Density Functional Theory (DFT).The synthesized material inhibits the human colon cancer cell line HT-29, the human lung adenocarcinoma cell line A549, and the human gastric cancer cell line MKN45, with substantial biological effects that have been demonstrated to be effective. According to in vitro and in sillico anticancer studies, PZ-PSBA has better activity against the human cervical cancer cell line (HeLa) and in bioinformatics analysis. PZ-PSBA suppressed pathogenic strains better than its parent chemical and can thus be suggested for the treatment of bacterial infections.

NXPH4 can be used as a biomarker for pan-cancer and promotes colon cancer progression.

NXPH4 promotes cancer proliferation and invasion. However, its specific role and mechanism in cancer remain unclear. Transcriptome and clinical data for pan-cancer were derived from the TCGA database. K-M survival curve and univariate Cox were used for prognostic analysis. CIBERSORT and TIMER algorithms were employed to calculate immune cell infiltration. Gene set enrichment analysis (GSEA) was employed for investigating the function of NXPH4. Western blot verified differential expression of NXPH4 in colon cancer. Functional assays (CCK-8, plate clonogenicity assay, wound healing assay, and Transwell assay) confirmed the impact of NXPH4 on proliferation, invasion, and migration of colon cancer cells. Dysregulation of NXPH4 in pan-cancer suggests its potential as a diagnostic and prognostic marker for certain cancers, including colon and liver cancer. High expression of NXPH4 in pan-cancer might be associated with the increase in copy number and hypomethylation. NXPH4 expression in pan-cancer is substantially linked to immune cell infiltration in the immune microenvironment. NXPH4 expression is associated with the susceptibility to immunotherapy and chemotherapy. Western blot further confirmed the higher expression of NXPH4 in colon cancer. Knockdown of NXPH4 significantly suppresses proliferation, invasion, and migration of colon cancer cell lines HT-29 and HCT116, as validated by functional assays.

Anticancer, antioxidant, and antibacterial activity of chemically fingerprinted extract from Cyclamen persicum Mill.

In the last few decades, researchers have thoroughly studied the use of plants in Palestine, one of them is Cyclamen persicum Mill. (C. persicum). Cyclamen persicum has been historically cultivated since the 1700s due to its tuber. The tuber is known to stimulate the nasal receptors, thus triggering the sensory neurons. Cyclamen persicum has anti-inflammatory effects, reduces cholesterol levels, treats diabetes, and inhibits tumor growth. In this respect, in-vitro examination of antibacterial and anticancer activities and antioxidative potency of C. persicum ethanolic extract were evaluated. The antioxidative potency of the extracted plant material was determined spectrophotometrically using the DPPH free radical scavenging method and the HPLC–PDA method to evaluate its total phenolic content (TPC) and total flavonoid content (TFC). The experimental results demonstrated limited antibacterial activity of C. persicum extract against both gram negative (E. coli) and gram positive (Streptococcus and S. aureus) bacterial strains, with the observed zones of inhibition measuring less than 8 mm. On the other hand, powerful activity against MCF7 breast cancer as well as HT29 colon cancer cell lines was obtained. The findings also revealed potent inhibition of free radicals and the presence of maximal levels of natural products such as phenolic compounds and flavonoids, which supportits biological activities and powerful ability to scavenge free radicals. HPLC results showed the presence of numerous flavonoid and phenolic compounds such as rutin, chlorogenic acid, kaempferol, trans-cinnamic acid, quercetin, sinapic acid, and p-coumaric acid.

10-Secocycloartane (=9,19-cyclo-9,10-secolanostane) triterpenoid saponins: Huangqiyenins M–X from Astragalus membranaceus (Fisch.) Bge

Phytochemical investigations of the leaves of Astragalus membranaceus (Fisch.) Bge. have led to the isolation of 12 undescribed triterpenoid saponins named huangqiyenins M–X. The structures of the undescribed compounds were determined using NMR and HRESIMS data. The cytotoxicity of these compounds against the RKO and HT-29 colon cancer cell lines was evaluated. Among these compounds, huangqiyenin W exhibited the highest cytotoxic activity against RKO colon cancer cells, whereas huangqiyenin Q and W showed moderate cytotoxic activity against HT-29 colon cancer cells. The network pharmacology results indicated that STAT3, IL-2 and CXCR1 are the correlated targets of huangqiyenin W against colon cancer, with AGE–RAGE and Th17 cell differentiation as the key signaling pathways.

Abstract 2608: Hu2G10 antibody-drug conjugates selectively target cleaved-activated Trop-2 in cancer cells and show therapeutic efficacy against multiple human cancers

Abstract Trop-2 is a transmembrane calcium signal transducer which activates growth-signaling networks that converge on Akt, ERK, Cyclin D1, NFkB (1). Trop-2 cleavage by ADAM10 was shown to be an activator switch for induction of cancer growth and metastatic diffusion (2). We devised a strategy to obtain monoclonal antibodies (mAb) capable of recognizing ADAM10-exposed sites in cancer cells. We succeeded at obtaining mAb that efficiently bound Trop-2 expressing cancer cells and were able to inhibit cell growth in vitro. We humanized the 2G10 mAb as the IgG1/k Hu2G10 antibody. Hu2G10 binds cancer-specific, cleaved/activated Trop-2 with Kd 10−12 M. On the other hand, it binds uncleaved/wtTrop-2 in normal cells with Kd 3.16 × 10−8 M, thus promising an unprecedented therapeutic index in patients. Antibody-drug conjugates (ADC) of Hu2G10 were obtained using stable linkers, multiple conjugation chemistries and different payloads, among them topoisomerase I inhibitors, bleomycin, RNA polymerase inhibitor, MMAE and calicheamicin. Hu2G10-ADC selectively killed Trop-2-expressing cells in vitro. Dose-response curves were obtained for the BxPC-3 pancreatic cancer, DU-145 prostate cancer, HT29 colon cancer cell lines and KM12SM/Trop2 colon cancer transfectants, reaching EC50 values as low as 5.05 × 10−10 M. Hu2G10-ADC drove high anticancer effectiveness on cell- and patient-derived xenografts from colon, pancreas, prostate and ovarian cancer. Effectiveness followed a gradient of payload potency, with correspondingly high therapeutic indexes. Consistently, neither detectable toxic effects nor weight loss were observed in any subgroup of treated xenotransplant-bearing mice. Taken together these findings candidate anti-Trop-2 Hu2G10-ADC for best-in-class anticancer therapy. (1) E. Guerra et al., The Trop-2 signaling network in cancer growth. Oncogene 32, 1594-1600 (2013).(2) M. Trerotola et al., Trop-2 cleavage by ADAM10 is an activator switch for cancer growth and metastasis. Neoplasia 23, 415-428 (2021). Citation Format: Saverio Alberti, Marco Trerotola, Antonino Moschella, Raffaella Giavazzi, Emanuela Guerra. Hu2G10 antibody-drug conjugates selectively target cleaved-activated Trop-2 in cancer cells and show therapeutic efficacy against multiple human cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2608.

Optimization of Aqueous Extraction of Polyphenols from Cuminum cyminum Seeds Using Response Surface Methodology and Assessment of Biological Activity.

(1) Background: Cumin seeds, extracted from the plant Cuminum cyminum, are abundant in phenolic compounds and have been extensively researched for their chemical makeup and biological effects. The objective of this research is to enhance the water extraction of polyphenols through the water bath (WB) technique and to evaluate the antiradical, antibacterial, and anticancer effects of the extract. (2) Methods: Response Surface Methodology was used to find the best parameters to extract polyphenols. Three experimental parameters, time, temperature, and solid-liquid ratio, were tested. The disc diffusion method has been used to determine the antimicrobial activities against Salmonella Typhimurium, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, and Candida albicans. The antiradical activity was performed using the DPPH method, while total phenolic content was performed using Folin-Ciocalteu. High-Performance Liquid Chromatography (HPLC) was conducted to analyze the phytochemical profile of WB extracts. The anticancer activity of the lyophilized extract was assessed against three cancer cell lines (colon (HT29), lung (A549), and breast (MCF7) cancer cell lines).; (3) Results: The optimal conditions for water extraction were 130 min at 72 °C. The total phenolic compounds yield (14.7 mg GAE/g DM) and antioxidant activity (0.52 mg trolox eq./mL) were obtained using a 1:40 solid-liquid ratio. The primary polyphenols identified were the flavonoids rutin (0.1 ppm) and ellagic acid (3.78 ppm). The extract had no antibacterial or antifungal activities against the microorganisms tested. The extract showed anticancer activity of about 98% against MCF7 (breast cancer cell line), about 81% against HT29 (colon cancer cell line), and 85% against A549 (lung cancer cell line) at high doses. (4) Conclusions: Extraction time and a high solid-liquid ratio had a positive impact on polyphenol recovery and in maintaining their quantity and quality. Furthermore, the optimal aqueous extract exhibited strong antiradical activity reflected by the inhibition of free radicals in addition to a significant specificity against the tested cancer cell lines.