HT-29 Colon Cancer Cells Research Articles

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.

Effects of Viable and Heat-Inactivated Bifidobacterium longum D42 on Proliferation and Apoptosis of HT-29 Human Colon Cancer Cells.

Bifidobacterium longum is a common probiotic; both viable and heat-inactivated Bifidobacterium longum have many probiotic effects, such as anticancer effects. But some mechanisms of anticancer effects are still unclear, especially for heat-inactivated probiotics. In this study, we analyzed the effects of viable and heat-inactivated Bifidobacterium longum D42 on human colon cancer cells (HT-29). Cell proliferation, membrane permeability and apoptosis were detected by using the CCK-8 method, LDH method and Annexin V-FITC/PI kits. The ROS level and mitochondrial membrane potential were examined using the fluorescent probes DCFH-DA and JC-1. Real-time fluorescence quantitative PCR (RT-qPCR) and Western blot were used to detect the expression of mitochondrial apoptosis pathway genes and proteins. The results showed that viable and heat-inactivated Bifidobacterium longum D42 at concentrations of 1 × 106 CFU/mL significantly inhibited the proliferation of and increased the level of LDH release of HT-29 colon cancer cells. We found that they could increase the apoptosis rate of HT-29 cells. Moreover, they could also induce apoptosis by inducing cells to produce ROS and destroying the mitochondrial membrane potential of cells. Further studies found that they could increase the mRNA transcription and protein expression levels of the Caspase-3, Caspase-9 and Bax genes in cells, and reduce the mRNA transcription and protein expression levels of the Bcl-2 gene. In summary, our findings revealed that viable and heat-inactivated Bifidobacterium longum D42 have inhibitory effects on proliferation and promote the apoptosis of human colon cancer cells, and also have certain adjuvant drug therapeutic effects and have potential application value in the adjuvant treatment of colon cancer.

Ulva lactuca L. extract bio-templated fabrication of CuO/ZnFe2O4 nanocomposites for photodegradation organic dye pollutants and in vitro biological activities

In this present work, a novel CuO/ZnFe2O4 nanocomposites has been fabricated via hydrothermal-assisted synthesis using Ulva lactuca L. extract as both a reducing and capping agent A comprehensive investigation such as structural, morphological, optical, and electronic properties of the CuO/ZnFe2O4 NCs were investigated in details. The photocatalytic efficacy of the prepared CuO/ZnFe2O4 NCs was assessed for Congo red (CR) dye degradation and the experimental results demonstrate the CuO/ZnFe2O4 NCs exhibits superior photocatalytic activity (91.13% k=0.03289 min−1) compared to pure CuO and ZnFe2O4 catalysts under 120 min of visible light irradiation. Additionally, the CuO/ZnFe2O4 NCs exhibited excellent long-term recycling stability over four consecutive cycles. This enhanced photocatalytic performance of CuO/ZnFe2O4 NCs was attributed to the synergism between CuO and ZnFe2O4, which facilitate the exceptional properties such as large specific area, a wide visible-light absorption range, and efficient electron-hole separation. Furthermore, the prepared CuO/ZnFe2O4 NCs were evaluated for their in vitro antibacterial and anticancer properties. The results of the disc diffusion assay with CuO/ZnFe2O4 NCs as an antibacterial agent demonstrated a highest zone of inhibition of 18.23 ± 0.30 mm for Bacillus subtilis and 19.83 ± 0.76 mm Escherichia coli. Moreover, in vitro cytotoxicity assessments using the MTT assay indicated a 64.94% cytotoxic activity against HT-29 colon cancer cells. The multifunctional advantages of Ulva lactuca L. extract-mediated green CuO/ZnFe2O4 NCs hold significant promise for environmental remediation and in vitro-based therapeutic applications.

Synthesis, characterisation, and anticancer and antioxidant activities of novel complexes of palladium and an organic Schiff-base ligand

Treatment of the Schiff base 2-N-(salicylidene) benzothiazole (SabtH) with palladium chloride afforded the neutral 2-N-(salicylidene) benzothiazole palladium(II) complex. This was then treated with one molar equivalent of either ethylenediamine or 1,10-phenanthroline, yielding the neutral mixed ligand complexes 1 and 2, respectively. The prepared organic ligand and its mixed ligand complexes were characterised via UV-Vis, FTIR, 1H-NMR, and 13C-NMR spectroscopy. The antiproliferative activities of 1 and 2 were tested against MCF-7 (breast) and HT-29 (colon) cancer cells using an MTT assay. 1 showed strong antiproliferative activity against both MCF-7 and HT-29 cancer cells, with IC50 values of 74.6 ± 8.35 and 174 ± 20.28 µg/mL, respectively. In addition, 2 exhibited the maximum antioxidant activity, with an EC50 value of 1029 ± 196.76 µg/mL.
 KEY WORDS: 2-N-(Salicylidene) benzothiazole, 1,10-Phenanthroline, Palladium mixed ligand complexes, Anticancer, Antioxidant activity
 Bull. Chem. Soc. Ethiop. 2024, 38(3), 605-613. 
 DOI: https://dx.doi.org/10.4314/bcse.v38i3.5

Preparation, characterization and in vitro evaluation of 5-fluorouracil loaded into chitosan-acacia gum nanoparticles.

Aim: In this study, we prepared, characterized and in vitro evaluated a 5-fluorouracil (5-FU)-loaded chitosan-acacia gum nanoparticles. Methods: Nanoparticles were characterized for their size, charge, morphology and encapsulation efficiency (EE%) followed by cellular investigations against HT-29 colon cancer cell line. Results: The nanoparticles exhibited a spherical morphological size with 94.42% EE%. Free 5-FU showed a fast and fully cumulative release after 6h while 5-FU loaded into CS-AG NPs showed good entrapment and slow, prolonged 5-FU release even after 24h. Enhanced IC50 for the 5-FU loaded NPs compared with free 5-FU against HT-29 colon cancer cell line was reported with high selectivity compared with normal fibroblast cells. Conclusion: 5-FU loaded NPs is promising nano-therapy against colon cancer.

Reliable quality of R-phycoerythrin derived from Portieria hornemannii for effective antioxidant, antibacterial, and anticancer activity

The phycobiliprotein is a light-harvesting fluorescent probe; Phycoerythrin is a pigment used in many industrial, and pharmaceutical applications; including fluorescence resonance energy transfer and flow cytometry. The main objective of the current study was purifying the phycoerythrin in Portieria hornemannii. The phycoerythrin pigment was dialyzed commonly using Q sepharose, but it is high cost. However, the alternative method is DEAE-Sephadex is cost-effective, high- yield, pure compound, and has high activity performance. Then the R-Phycoerythrin (R-PE) compound was extracted in 0.02 M of phosphate buffer (pH–7.2). R-PE investigated the FT-IR, HPLC, SDS-PAGE and including physico-chemical properties. Then the purified R-PE compound was evaluated by an antibacterial, antioxidant, and anticancer activity. The R-Phycoerythrin was showing a maximum inhibition effect of high concentration (IC50value 77.98 µg/ml) against the colon cancer HT-29 cell line.

Diterpenoids from Torreya grandis and their cytotoxic activities

Eight previously undescribed diterpenoids, along with eleven previously reported analogues, were obtained from the supercritical CO2 extracts of Torreya grandis aril. The structures of these compounds were elucidated based on HRESIMS, NMR, ECD, and single-crystal X-ray diffraction data. In the MTT assay, compound 18 exhibited significant inhibitory effects on two human colon cancer cell lines, HT-29 and HCT 116 cells, with IC50 values of 7.37 μM and 6.55 μM, respectively. It was found that compound 18 induced apoptosis and significantly inhibited the migration of HCT 116 colon cancer cells in a concentration-dependent manner.

Nanoliposome-loaded phenolics from Salvia leriifolia Benth and its anticancer effects against induced colorectal cancer in mice.

Colon cancer is one of the leading causes of death among various types of cancer. Despite the significant progress made in cancer treatment, chemotherapy resistance and various side effects are still prevalent. The objective of this study is to assess the therapeutic potential of phenolic-rich fraction encapsulated nanoliposome (PRF-NLs) of Salvia leriifolia Benth in the treatment of colon cancer in mice. Initially, the phenolic-rich fraction (PRF) was extracted and then encapsulated into nanoliposomes. The physicochemical properties of the nanoliposomes were evaluated using dynamic light scattering, zeta potential, and field emission scanning electron microscopy. Subsequently, 24 mice with HT-29 colon cancer cells were divided into three groups, and the anticancer effects of PRF-NLs were measured. The results showed that the ethyl acetate fraction of S. leriifolia was the highest PRF containing 14.27 ± 2.39mg (gallic acid) GA/g DW (dry weight), and the PRF successfully loaded into the nanoliposome structure resulted in the synthesis of nanoliposomes with a nanometer size and spherical shape and homogenous dispersion. Some of the abundant bioactive phenolic compounds in the nanoliposome-loaded PRF are salicylic acid and naringin. The average daily weight gain and food intake, and changes in the expression of caspase 3, Bax (Bcl-2 associated X-protein), and Bcl2 (B-cell lymphoma 2), inducible nitric oxide synthase genes, were observed in the mice group induced colorectal cancer cells. At a dose of 100mg TPC (total phenolic content)/kg BW/day, the nonencapsulated PRF dietary addition improved these parameters; however, the potential shown by nanoliposome-encapsulated PRF than the nonencapsulated PRF in enhancing health parameters in mice was higher. The developed intestinal absorption and bioavailability of nanoliposome-encapsulated PRF contribute to its increased health-promoting activity. Thereby, the synthesized nanoliposome may be a potential natural anticancer drug to prevent colorectal cancer.

Rheum khorasanicum. Hydroalcoholic root extract induces cell death in human colorectal adenocarcinoma: An in vitro and in silico study

Colorectal cancer (CRC) is the second greatest cause of cancer-related death in the world and chemotherapy, as an important part of CRC treatment, has some drawbacks, including systemic toxicity. Therefore, it is crucial to discover new and more effective CRC treatment plans. Rheum khorasanicum (R.khorasanicum) is a medicinal plant with high flavonoids, stilbenes, and anthraquinone contents, so it can be a potential source of antioxidants and can be used for therapeutic purposes and trigger apoptosis in cancer cells. In this study, we investigated the effects of hydroalcoholic root extract of R.khorasanicum treatment on inducing mitochondrial apoptosis of HT-29 and Caco-2 human colorectal adenocarcinoma cells. Firstly, the total phenolic and flavonoid content was determined. Then, the cytotoxic effects of R.khorasanicum on cells of three different types, including HT-29 and Caco-2 colon cancer cells as well as normal 3T3 cells were assessed using the MTT assay. To investigate the characteristics of cellular death, flow cytometry, and western blotting were performed. The results of this study indicated considerable phenolic (356.4±9.4 GAE/gDW) and flavonoid (934.55±17.1 QE/gDW) contents in R.khorasanicum. MTT assay's finding indicated that 100, 60, and 30μg/mL concentrations of R.khorasanicum reduce cell viability in HT-29 and Caco-2 cell lines significantly (P<0.05). It has been also revealed that R.khorasanicum extract induces apoptosis rather than necrosis in these cell lines. Moreover, Bcl-2 expression was significantly reduced in both HT-29 and Caco-2 cell lines, while Bax and cleaved caspase-3 expression soared considerably in the groups under R.khorasanicum treatment (P<0.05). In conclusion, our findings have suggested that high phenol and flavonoid contents of R.khorasanicum root extract possibly play an important role in cell cytotoxicity and apoptosis induction in HT-29 and Caco-2 colon cancer cells.

Fabrication of poly (lactic-co-glycolic acid)/mesoporous silica composite nanofibers for controllable co-delivery of 5-fluorouracil and curcumin against HT-29 colon cancer cells

Fabrication of poly (lactic-co-glycolic acid)/mesoporous silica composite nanofibers for controllable co-delivery of 5-fluorouracil and curcumin against HT-29 colon cancer cells

5-Fluorouracil-Loaded PLGA Declined Expression of Pro-Inflammatory Genes IL-9, IL-17A, IL-23 and IFN- y; in the HT-29 Colon Cancer Cell Line.

Pro-inflammatory cytokines play critical roles in cancer pathobiology and have been considered potential targets for cancer management and therapy. Understanding the impact of cancer therapeutics such as 5-fluorouracil (5-FU) on their expression might shed light on development of novel combinational therapies. This study aimed to encapsulate 5-FU into PLGA and evaluate their effects on the expression of pro-inflammatory genes IL-9, IL-17-A, IL-23, and IFN-y; in the HT-29 cells. PLGA-5-FU NPs were constructed and characterized by Dynamic Light Scattering (DLS) and Atomic Force Microscopy (AFM). The cytotoxicity was evaluated by MTT test and, the IC50 was identified. HT-29 cells were treated with different concentrations of the PLGA-5-FU NPs for 48 hours and, gene expression levels were analyzed by qRT-PCR. DLS and AFM analysis revealed that the prepared PLGA-5-FU NPs were negatively charged spherical-shaped particles with a mean size of 215.9 ± 43.3 nm. PLGA-5-FU NPs impacted the viability of HT-29 cells in a dose- and time-dependent manner. The qRT-PCR results revealed a dose-dependent decrease in the expression of IL-9, IL-17A, IL-23 and IFN-y; genes, and their expressions were significantly different in both 10 and 20 µg/mL treated groups compared to the control. However, although the treatment of HT-29 cells with 20 µg/mL free 5-FU resulted in decreased expression of the studied genes, the differences were not statistically significant compared to the control group. PLGA-5-FU NPs significantly suppressed expression of the IL-9, IL-17A, IL-23 and IFN-y; genes, and the encapsulation of 5-FU into PLGA improved considerably impact of the 5-FU on the HT-29 cells.

Caffeic acid phenethyl ester mediates apoptosis in serum-starved HT29 colon cancer cells through modulation of heat shock proteins and MAPK pathways.

Colorectal cancer (CRC) is among the most prevalent gastrointestinal cancers of epithelial origin worldwide, with over 2 million cases detected every year. Emerging evidence suggests a significant increase in the levels of inflammatory and stress-related markers in patients with CRC, indicating that oxidative stress and lipid peroxidation may influence signalling cascades involved in the progression of the disease. However, the precise molecular and cellular basis underlying CRC and their modulations during bioactive compound exposure have not yet been deciphered. This study examines the effect of caffeic acid phenethyl ester (CAPE), a natural bioactive compound, in HT29 CRC cells grown under serum-supplemented and serum-deprived conditions. We found that CAPE inhibited cell cycle progression in the G2/M phase and induced apoptosis. Migration assay confirmed that CAPE repressed cancer invasiveness. Protein localisation by immunofluorescence microscopy and protein expression by western blot analysis reveal increased expressions of key inflammatory signalling mediators such as p38α, Jun N-terminal kinase and extracellular signal-regulated kinase (ERK) proteins. Molecular docking data demonstrates that CAPE shows a higher docking score of -5.35 versus -4.59 to known p38 inhibitor SB203580 as well as a docking score of -4.17 versus -3.86 to known ERK1/2 inhibitor AZD0364. Co-immunoprecipitation data reveals that CAPE treatment effectively downregulates heat shock protein (HSP) expression in both sera-supplemented and limited conditions through its interaction with mitogen-activated protein kinase 14 (MAPK14). These results suggest that stress induction via serum starvation in HT29 CRC cells leads to the induction of apoptosis and co-ordinated activation of MAPK-HSP pathways. Molecular docking studies support that CAPE could serve as an effective inhibitor to target p38 and MAPK compared to their currently known inhibitors.

The Cytotoxic Activity of Marine Sponge-Derived Fungus Aspergillus nomius NC06 Against HT29 Colon Cancer Cells

The study of natural products from marine-derived fungi has been interesting tense to researchers as drug discovery sources. Marine fungus from West Sumatera, Indonesia repeatedly showed their potential for cytotoxic and antimicrobial activities. This study aims to determine the cytotoxic activity against HT29 colon cancer cells of each fraction of ethyl acetate extracts from Aspergillus nomius NC06 derived from marine sponge Neopetrosia chaliniformis. A. nomius was cultivated with rice as a growth medium and extracted with ethyl acetate solvent and evaporated in vacuo to obtain ethyl acetate extract. Furthermore, the compounds of ethyl acetate extract were separated with the VLC (Vacuum Liquide Chromatography) method. Five fractions were obtained, which further investigated their cytotoxic activity against HT29 colon cancer cells by using an MTT assay. The result showed that fractions I and III were categorized as potential fractions due to their IC50 value of 13.12 ± 0.39 μg/mL and 2.59 ± 0.19 μg/mL, respectively. It was also supported by ANOVA to measure the effect of each concentration (0.1; 1; 10; 100 μg/mL) of each fraction on the viability percentage of HT29 cells with p &lt; 0.005.

Designing nanostructured lipid carriers modified with folate-conjugated chitosan for targeted delivery of osthole to HT-29 colon cancer cells: investigation of anticancer, antioxidant, and antibacterial activities

AbstractThe present study proposed to design nanostructured lipid carriers (NLC) coated with chitosan (CS) conjugated folate (FA) for the targeted delivery of Osthole (OST) to the HT-29 colon cancer cell line and improve its anticancer capability. To assess the physicochemical characteristics of OST-loaded NLC decorated with CS-conjugated FA (OST-NCF-NPS), several techniques, including DLS, SEM, and FTIR, were applied. After determining the encapsulation efficiency of OST in CSFA-modified NLC-NPs, an MTT test was conducted to evaluate the cytotoxic effects of this nano platform on the HT-29 cancer cell line in comparison to normal HFF cells. Possible mechanisms of apoptosis in cancer cells treated with OST-NCF-NPs were examined using qPCR, flow cytometry, and AO/PI fluorescent staining methods. Moreover, the antioxidant capacity of these biosynthesized nanocarriers was determined using ABTS and DPPH methods, and their antibacterial potential was measured through disk diffusion, MIC, and MBC assays. According to the findings, OST-NCF-NPS had the ideal average size of 179.19 nm, low polydispersity (PI = 0.23), acceptable physical stability (ζ-potential = + 18.99 mV), and high entrapment efficiency (83.5%). The MTT data demonstrated the selective cytotoxicity of NPs toward cancerous cells compared to normal cells. Cell cycle and Annexin V/Propidium Iodide (AnV/PI) analysis indicated that OST-NCF-NPs increased the sub-G1 population and AnV/PI-positive cells. The occurrence of programmed cell death in the treated cells was also verified by altered expression of proapoptotic (BAX and caspase-3) and antiapoptotic (Bcl-2) genes. Furthermore, the NPs exhibited strong antibacterial activity, particularly against gram-negative bacteria, and high antioxidant effects in reducing ABTS and DPPH-free radicals. Graphical Abstract

Analyzing of colorectal cancerrelated genes and microRNAs expression profiles in response to probiotics Lactobacillus acidophilus and Saccharomyces cerevisiae in colon cancer cell lines.

Colorectal cancer is the world's third most frequent cancer and the fourth cause of mortality. Probiotics play an important function in preventing metastasis as well as the growth and proliferation of malignant cancer cells. The study investigated the anticancer effect of Lactobacillus acidophilus supernatant and Saccharomyces cerevisiae yeast on colorectal cell lines, including HT29 and SW480 as a colorectal cancer model. The extract from the Lactobacillus acidophilus and Saccharomyces cerevisiae standard probiotics were prepared, and probiotics characterization was confirmed by morphological and Biochemical tests. The viability of HT29 and SW480 colon cancer cell lines on effecting probiotic supernatant was evaluated by measuring the MTT colorimetric method. Comparison of the expression profile of several genes involved in apoptosis, cell cycle, and metastatic pathway in HT29 and SW480 cell lines with the treatment of probiotics extract showed an upregulation in the BAX, CASP3, and CASP9 and down regulation BCl-2, MMP2, and MMP9 genes. Also, a comparison of microRNA expression profiles indicated an increase of miR 34, 135, 25, 16, 195, 27, 98, let7 and a decrease of miR 9, 106b, 17, 21, 155, 221. The findings of this study indicate that probiotics can effectively suppress the proliferation of colorectal cancer cells and even reverse their development. Additionally, the study of cellular genes and miRNA profiles associated with colorectal cancer have demonstrated that our probiotics play a crucial role in CRC prevention by increasing the expression of tumor suppressor microRNAs and their target genes while decreasing oncogenes.

Effects of Quercetin Treatment on Apoptosis and Ferroptosis Cell Death Mechanisms in Colon Cancer Cell Lines

One of the key challenges in cancer research is finding ways to selectively eliminate cancer cells without harming healthy tissues. This makes understanding cell death mechanisms in cancer cells critical for developing effective treatments. Ferroptosis, a recently recognized form of programmed cell death driven by iron, has emerged as a promising approach for targeting cancer cells, which have higher iron requirements than normal cells. Inducing ferroptosis has been shown to overcome resistance to various chemotherapeutic agents and targeted therapies, enhancing their effectiveness and decreasing cancer resistance. To explore the protective effects of quercetin, a flavonoid metabolite, and the cell death pathways it engages, experiments were conducted using HT-29 and HCT-116 colon cancer cell lines treated with quercetin and an anticancer drug. Cell viability was measured after 24 and 48 hours of incubation with varying logarithmic concentrations of quercetin and doxorubicin, both independently and in combination. The study focused on identifying the cell death pathways activated by these treatments by analyzing apoptosis markers APAF-1 and CASP-3, along with ferroptosis markers GPx4 and ACSL-4. The results indicated that quercetin alone elevated APAF-1 and CASP-3 levels in both colon cancer cell lines, thus promoting the apoptotic pathway. In terms of ferroptosis, the combination treatment increased ACSL-4 expression while reducing GPx4, indicating the induction of ferroptosis and a synergistic effect on cancer cells. These findings point to the potential for further investigation into combining ferroptosis-inducing agents and the signaling pathways that trigger different forms of cell death.

The effect of curcumin on the necroptosis signaling pathway in colon cancer cells

Colon cancer is the one of the most common types of cancer in humans. A sedentary lifestyle, increasing obesity and the consumption of food additives favor the development and occurrence of colon cancer. It is emphasized that curcumin, a yellow compound isolated from the turmeric plant, is important in preventing cancer. Studies have shown that curcumin has an anticancer effect by driving cancer cells into apoptosis, but studies showing its effect on necroptosis are inconclusive. Necroptosis is a form of programmed cell death mediated by RIP proteins and has been shown to play an important role in cancer. This study aims to determine the effect of curcumin on the necroptosis signaling pathway. For this purpose, HT-29 and HCT-116 colon cancer cells were cultured and exposed to different concentrations of curcumin and MTT experiments were performed to determine the effect on cell viability. The expression levels of RIPK1, RIPK3, and MLKL genes, which are markers of necroptosis, were analyzed by real-time PCR. It was found that the expression level of RIPK1, RIPK3, and MLKL genes significantly increased after exposure of HT -29 cells to 50 µM curcumin. Moreover, the expression of RIPK1 and MLKL genes increased in HCT-116 cells after curcumin administration. Consequently, the current data clearly suggest that curcumin is a prominent driver of necroptotic signaling-mediated colon cancer cell death.

Exploring the Bioactive Potential of Calostoma insigne, an Endangered Culinary Puffball Mushroom, from Northeastern Thailand.

Calostoma insigne puffball mushrooms are only found in forests with rich biodiversity in very few countries including Thailand, and their biofunctions remain largely unexplored. This study used the agar disk diffusion assay, the anti-glucosidase assay, and the 3, 4, 5-dimethylthiazol-2-yl-2-5-diphenyltetrazolium bromide (MTT) assay to evaluate the bioactive potential of these endangered puffball mushrooms. Internal transcribed spacer (ITS) gene analysis identified C. insigne, a puffball mushroom with green, globose, and spiny spores. Fourier-transform infrared spectroscopy (FTIR) analysis confirmed the polysaccharide structure while scanning electron microscopy (SEM) revealed a fiber-like network. The ethanolic gelatinous fruiting body extract exhibited 1,1-diphenyl-2-picrylhydrazyl (DPPH)-scavenging capacity (57.96%), a ferric ion-reducing antioxidant power (FRAP) value of 1.73 mg FeSO4/g, and α-glucosidase inhibition (73.18%). C. insigne cytotoxicity was effective towards HT-29 colon cancer cells using the MTT assay (IC50 of 770.6 µg/mL at 72 h) and also showed antiproliferative capacity (IC50 of 297.1 µg/mL). This puffball mushroom stimulated apoptotic genes and proteins (caspase-3, Bax, and p21) via an intrinsic apoptotic pathway in HT-29 cells. In the laboratory, the medium formula consisting of 20% potato, 2% sucrose, and 0.2% peptone was optimal to increase fungal mycelial biomass (2.74 g DW/100 mL), with propagation at pH 5.0 and 30 °C. Puffball mushrooms are consumed as local foods and also confer several potential health benefits, making them worthy of conservation for sustainable utilization.

New pyrrolidine-carboxamide derivatives as dual antiproliferative EGFR/CDK2 inhibitors.

Cancer is one of the leading causes of mortality worldwide, making it a public health concern. A novel series of pyrrolidine-carboxamide derivatives 7a-q were developed and examined in a cell viability assay utilizing a human mammary gland epithelial cell line (MCF-10A), where all the compounds exhibited no cytotoxic effects and more than 85% cell viability at a concentration of 50 μM. Antiproliferative activity was evaluated invitro against four panels of cancer cell lines A-549, MCF-7, Panc-1, and HT-29. Compounds 7e, 7g, 7k, 7n, and 7o were the most active as antiproliferative agents capable of triggering apoptosis. Compound 7g was the most potent of all the derivatives, with a mean IC50 of0.90 μM compared to IC50 of 1.10 μM for doxorubicin. Compound 7g inhibited A-549 (epithelial cancer cell line), MCF-7 (breast cancer cell line), and HT-29 (colon cancer cell line) more efficiently than doxorubicin. EGFR inhibitory assay results of 7e, 7g, 7k, 7n, and 7o demonstrated that the tested compounds inhibited EGFR with IC50 values ranging from 87 to 107 nM in comparison with the reference drug erlotinib (IC50 = 80 nM). 7e, 7g, 7k, 7n, and 7o inhibited CDK2 efficiently in comparison to the reference dinaciclib (IC50 = 20 nM), with IC50 values ranging from 15 to 31 nM. The results of inhibitory activity assay against different CDK isoforms revealed that the tested compounds had preferential inhibitory activity against the CDK2 isoform.