HCT116 Colon Cancer Cells Research Articles

The Inhibitory Effect and Mechanism of the Histidine-Rich Peptide rAj-HRP from Apostichopus japonicus on Human Colon Cancer HCT116 Cells

Colon cancer is a common and lethal malignancy, ranking second in global cancer-related mortality, highlighting the urgent need for novel targeted therapies. The sea cucumber (Apostichopus japonicus) is a marine organism known for its medicinal properties. After conducting a bioinformatics analysis of the cDNA library of Apostichopus japonicus, we found and cloned a cDNA sequence encoding histidine-rich peptides, and the recombinant peptide was named rAj-HRP. Human histidine-rich peptides are known for their anti-cancer properties, raising questions as to whether rAj-HRP might exhibit similar effects. To investigate whether rAj-HRP can inhibit colon cancer, we used human colon cancer HCT116 cells as a model and studied the tumor suppressive activity in vitro and in vivo. The results showed that rAj-HRP inhibited HCT116 cell proliferation, migration, and adhesion to extracellular matrix (ECM) proteins in vitro. It also disrupted the cytoskeleton and induced apoptosis in these cells. In vivo, rAj-HRP significantly inhibited the growth of HCT116 tumors in BALB/c mice, reducing tumor volume and weight without affecting the body weight of the tumor-bearing mice. Western blot analysis showed that rAj-HRP inhibited HCT116 cell proliferation and induced apoptosis by upregulating BAX and promoting PARP zymogen degradation. Additionally, rAj-HRP inhibited HCT116 cell adhesion and migration by reducing MMP2 levels. Further research showed that rAj-HRP downregulated EGFR expression in HCT116 cells and inhibited key downstream molecules, including AKT, P-AKT, PLCγ, P38 MAPK, and c-Jun. In conclusion, rAj-HRP exhibits significant inhibitory effects on HCT116 cells in both in vitro and in vivo, primarily through the EGFR and apoptosis pathways. These findings suggest that rAj-HRP has the potential as a novel targeted therapy for colon cancer.

Hyaluronic acid-functionalized carboxymethyl dextran-coated melatonin nanoconjugates for targeted etoposide delivery in metastatic colon cancer: Extensive in-vitro investigation in HCT116 cell lines, antimicrobial efficacy, and anti-angiogenic potential in chick chorioallantoic membrane (CAM) assay

Managing advanced colon cancer is challenging, requiring targeted therapies. This study presents a novel nanoconjugate system, HA-CMD@ETP-MLT-NCs, designed to deliver etoposide (ETP) specifically to colon cancer cells. The system consists of Hyaluronic Acid (HA)-Functionalized Carboxymethyl Dextran (CMD) coated with Melatonin (MLT). The nanoconjugates showed good stability, with a zeta potential of −29.90 mV and a particle size of 199.1 nm. They achieved an 80.3 % yield and a high drug entrapment efficiency of 93.4 %. In vitro release studies demonstrated pH-dependent drug release, with 73.4 % released at pH 5.5 (tumour-like environment) and 42.6 % at pH 7.4 (normal tissue) over 24 h. The nanoconjugates improved cellular uptake, induced apoptosis, and reduced reactive oxygen species (ROS) in HCT116 colon cancer cells. Flow cytometry showed a significant decrease in ROS levels, and lipid peroxidation inhibition increased to 56.67 %. These findings suggest that HA-CMD@ETP-MLT-NCs enhance etoposide delivery and reduce side effects. Further in vivo studies and clinical trials are needed to confirm its therapeutic potential.

Activating transcription factor 4 plays a major role in shaping the transcriptional response to isoginkgetin in HCT116 cells

Activating transcription factor 4 (ATF4) plays a central role in the integrated stress response (ISR) and one overlapping branch of the unfolded protein response (UPR). We recently reported that the splicing inhibitor isoginkgetin (IGG) induced ATF4 protein along with several known ATF4-regulated transcripts in a response that resembled the ISR and UPR. However, the contribution of ATF4-dependent and -independent transcriptional responses to IGG exposure was not known. Here we used RNA-sequencing in HCT116 colon cancer cells and an isogenic subline lacking ATF4 to investigate the contribution of ATF4 to IGG-induced changes in gene expression. Approximately 85% of the IGG-responsive DEGs in HCT116 cells were also differentially expressed in response to the ER stressor thapsigargin (Tg) and these were enriched for genes associated with the UPR and ISR. Most of these were positively regulated by IGG with impaired responses in the ATF4-deficient cells. Nonetheless, there were DEGs that responded similarly in both cell lines. The ATF4-independent IGG-induced DEGs included several metal responsive transcripts encoding metallothionines and a zinc transporter. Taken together, the predominant IGG response was ATF4-dependent in these cells and resembled the UPR and ISR while a second less prominent response involved the ATF4-independent regulation of metal responsive mRNAs.

Integrated network pharmacology and transcriptomic approach reveal the role of equol in reducing colorectal cancer via regulating multiple cell cycle genes in HCT116 cells

Equol is an isoflavone-derived metabolite known to exhibit strong estrogenic and antioxidant activities. The aim of this paper is twofold: first, to confirm the anticancer potential of equol against colorectal cancer, and second, to reveal the underlying mechanisms. After treatment with 40 μg/mL equol, cell proliferation, cell migration, and colony formation of HCT116 colon cancer cells were inhibited. Network pharmacology and transcriptomics analysis revealed the downregulation of genes related to DNA replication (CCND1, E2F1, CDC6, CDC45, MCM4), leading to G1/S cell cycle arrest and the induction of cell apoptosis, which was confirmed by flow cytometry. Genes associated with the G2-to-M transition (CDK1, CCNA2, CCNB1) were also downregulated. In addition, equol downregulated genes (FOXM1 and ASPM) that control cell migration and invasion. Our data indicate that equol can inhibit colorectal cancer by targeting multiple pathways, suggesting its potential as a key component in the adjuvant treatment of colorectal cancer.

Antiproliferative and Apoptotic Effects of Solenostemma argel Leaf Extracts on Colon Cancer Cell Line HCT-116

Colorectal cancer ranks as the fourth most frequent cause of cancer-related fatalities on a global scale. The present study aims at assessing the anti-proliferative and apoptotic effects of Solenostemma argel extract on colorectal cancer cells (HCT-116). The antiproliferative activity was investigated using Sulfo-Rhodamine-B (SRB) assay and the apoptotic effects were demonstrated utilizing acridine orange/ ethidium bromide (AO/EB) staining method. The antiproliferative results demonstrated that the extract exhibited dose-dependent antiproliferative activity, with an IC50 value of 85.3 μg/ml. The apoptosis results clearly demonstrated the ability of the methanolic extract of Solenostemma argel in inducing apoptosis in HCT-116 cancer cells. In conclusion, the investigation highlights the considerable antiproliferative and apoptotic impacts of Solenostemma argel leaf extract on HCT-116 colorectal cancer cells. This underscores its potential as a promising chemopreventive agent specifically targeting HCT-116 colon cancer cells.

ANTI-CANCEROUS POTENTIAL OF POLYSACCHARIDES DERIVED FROM WHEAT CELL CULTURE

There is a global need to discover effective anti-cancerous compounds from natural sources. Cultivated wheat cells can be a valuable source of non-toxic or low toxic plant-derived polysaccharides. In this study we evaluated the antitumor ability of seven fractions of wheat cell culture polysaccharides (WCCPSs) in the HCT-116 colon cancer cell line. Almost all (6/7) fractions had an inhibitory effect on the proliferation of colon cancer cells, and two fractions (A-b and A-f) had considerable therapeutic indexes. The WCCPS fractions induced cell cycle arrest in the G1 phase and induced different rates of apoptosis (<48%). Transmission and scanning electron microscopy revealed that WCCPS fractions caused apoptotic changes in the nucleus and cytoplasm, including damage to mitochondria and external morphological signs of apoptosis. In addition, the WCCPSs induced an increase in the levels of Bax, cytochrome c, caspases 8 and 3, indicating that cell death was assessed through intrinsic and extrinsic pathways of apoptosis. Furthermore, some fractions caused a significant decrease of c-Myc, b-catenin, NFkB2 and HCAM (CD 44) levels, indicating enhanced cell differentiation. Thus, for the first time, our results provide a proof of concept of the anti-cancer capacity of WCCPS fractions in colorectal cancer. This research was supported by the FEDER Operational Program 2020/Junta de Andalucía-Consejería de Economía y Conocimiento/ Project (B-CTS-562-UGR20) and the Chair “Doctors Galera-Requena in cancer stem cell research” (CMC-CTS963); Erasmus+ Mobility Program; “Bolashak” Presidential Scholarship of the Republic of Kazakhstan for scientific work abroad; Scottish Government’s Rural and Environment Science and Analytical Services (RESAS) division.

Synthesis and photoluminescence properties of Ce3+/Tb3+ and Eu3+ ions doped GdB3O6/Ca10(PO4)6(OH)2 Core/Shell particles as well as its cytotoxicity against colon cancer cells

In this study, Ce3+/Tb3+ co-doped, Eu3+ doped GdB3O6 phosphor particles were synthesized by sol-gel method, coated with hydroxyapatite (HAP) and loaded with doxorubicin. Different chelating agents and surfactants (Cetyltrimethylammonium bromide (CTAB), citric acid, glycine, ethylenediaminetetraacetic acid (EDTA) and tartaric acid) were used in the synthesis in order to see their effect on the structure. While single phase Gadolinium triborate, GdB3O6 was obtained by using CTAB in precursor solution in the synthesis and Gadolinium orthoborate, GdBO3 produced when EDTA was used in precursor solution. Other organic molecules added to precursor solution produced mixtures of both phases. The highest photoluminescence intensity was obtained in the Gd0.95Ce0.025Tb0.025B3O6 and Gd0.825Eu0.175B3O6. In order to increase the drug loading efficiency, The Gd0.825Eu0.175B3O6 core was coated with Calcium hydroxyapatite, HAP. The core had an average size of 458.65 nm which increased to 616.85 nm after coating as seen with dynamic light scattering (DLS). Gd0.825Eu0.175B3O6@HAP particles were loaded with 7.82 % ± 0.80 wt doxorubicin and the release was studied at 37 °C in pH 7.4 phosphate-buffered saline (PBS) and pH 5.5 acetate buffer solutions. The total number of moles of drug released versus luminescence intensity was measured during the release studies and real-time imaging studies were carried out. Finally, cytotoxicity experiments were performed on HCT-116 colon cancer cells.

EP4 receptor agonist CAY10598 upregulates ROS-dependent Hsp90 cleavage in colorectal cancer cells

Prostaglandin E2 (PGE2) interacts with four specific G protein-coupled receptors, namely EP1, EP2, EP3, and EP4, playing a pivotal role in determining the fate of cells. Our previous findings highlighted that stimulating the EP4 receptor with its agonist, CAY10598, triggers apoptosis in colon cancer HCT116 cells via the production of reactive oxygen species (ROS). This process also reduces the phosphorylation of the oncogenic protein JAK2 and leads to its degradation in these cells. In this study, our goal was to explore the pathways through which CAY10598 leads to JAK2 degradation. We focused on Hsp90, a heat shock protein family member known for its role as a molecular chaperone maintaining the stability of several key proteins including EGFR, MET, Akt, and JAK2. Our results show that CAY10598 decreases the levels of client proteins of Hsp90 in HCT116 cells, an effect reversible by pretreatment with the ROS scavenger N-acetyl cysteine (NAC) or the proteasome inhibitor MG132, indicating that the degradation is likely driven by ROS. Furthermore, we observed that CAY10598 cleaves both α and β isoforms of Hsp90, the process inhibited by NAC. Inhibition of EP4 with the antagonist GW627368x not only prevented the degradation of Hsp90 client proteins but also the cleavage of Hsp90 itself in CAY10598-treated HCT116 cells. Additionally, CAY10598 suppressed the growth of HCT116 cells implanted in mice. Our findings reveal that CAY10598 induces apoptosis in cancer cells by a novel mechanism involving the ROS-dependent cleavage of Hsp90, thereby inhibiting the function of crucial Hsp90 client proteins.

The m6A methyltransferase METTL3 modifies Kcnk6 promoting on inflammation associated carcinogenesis is essential for colon homeostasis and defense system through histone lactylation dependent YTHDF2 binding

Inflammation induces tumor formation and plays a crucial role in tumor progression and prognosis. KCNK6, by regulating K(+) efflux to reduce NLRP3 Inflammasome-induced lung injury, relaxes the aorta. This study aims to elucidate the effects and biological mechanism of KCNK6 in inflammation-associated carcinogenesis, which may be essential for colon homeostasis and the defense system. To induce colitis, mice were given 3.0% Dextran Sodium Sulfate (DSS) in their drinking water for 7 days. The Azoxymethane (AOM) +DSS method was used to induce colon cancer in the mice model. Bone marrow-derived macrophages (BMDM) from Kcnk6-/- mice, AW264.7 cells, and human colon cancer HCT116 and Caco2 cells were used as in vitro models. The loss of Kcnk6 prevented spontaneous colitis and restored mucosal integrity and homeostatic molecules. Additionally, the loss of Kcnk6 reduced the severity of AOM/DSS-induced carcinogenesis. Kcnk6 promoted cell viability and proliferation in HCT-116 or Caco-2 cells. The loss of Kcnk6 inhibited the levels of inflammatory factors in BMDM cells. Kcnk6 accelerated potassium channel activity, inducing NLRP3 inflammasome activation. METTL3-mediated m6A modification increased Kcnk6 stability in a YTHDF2-dependent manner. Histone lactylation activated the transcription of YTHDF2/Kcnk6. Our study revealed the important role of Kcnk6 in inflammation-associated carcinogenesis progression. The m6A methyltransferase METTL3 and histone lactylation increased Kcnk6 stability in a YTHDF2-dependent manner, providing a potential strategy for inflammation-associated carcinogenesis or colorectal cancer therapy.

Enhancement of Cinnamomum camphora magnetic nanoparticle bioactivities via carboxymethyl cellulose immobilization for potential therapeutic applications in cancer treatment

A promising method for cancer therapy is the coating of magnetic nanoparticles with carboxy methylcellulose. In a research project, hydroalcoholic extract of Cinnamomum camphora leaves was used to demonstrate the production of magnetic nanoparticles (MNPs); MNPs were coated with carboxymethyl cellulose to form carboxymethyl cellulose-coated magnetic nanoparticles (CMNPs)were formed. Preliminary phytochemical screening of C. camphora confirmed the presence of flavonoids, carbohydrates, phenolic compounds, and proteins. Phenolics 280.59 (mg/g), flavonoids 15.46 (mg/g), proteins 1.9 (mg/mL) and total carbohydrates 293.80 (mg/g) were all quantified. To confirm the formation of MNPs and CMNPs, UV–visible (UV–vis) spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS) were used. Peaks were observed at 232 nm and 240 nm, respectively. The largest absorption peaks were observed in MNPs and CMNPs, respectively. The particles were spherical in shape and less than 10 (nm) in diameter. The potential scavenging activity of biosynthesized MNPs and CMNPs was evaluated by the ABTS and DPPH assays, and the inhibition values IC50 were 141.3 ± 3.0 and 61.67 ± 2.5 (µg/mL) for ABTS and 176.1 ± 4.0 and 70.92 ± 3.0 (µg/mL) for DPPH, respectively (p ≤ 0.05). Furthermore, the cytotoxicity test results showed that the HCT-116 human colon cancer cell line had the lowest IC50 value of 20 (µg/mL) for CMNP, followed by the HepG2 hepatocellular carcinoma cell line with an IC50 value of 33 (µg/mL) for CMNP, indicating that the cytotoxic effect on colon cancer cells is stronger than on liver cancer cells. Molecular docking studies have revealed that CMNPs target and bind to apoptotic protein, enhancing their bioactivity and cytotoxic effects on cancer cells. Furthermore, our findings suggest that the induction of apoptosis may be responsible for the anticancer effects of CMNPs.Graphical abstract

Effect of central metal ion on some pharmacological properties of new Schiff base complexes. Anticancer, antioxidant, kinetic/thermodynamic and computational studies

The biological capacities of Schiff Base complexes such as anti-cancer, anti-microbial and anti-oxidant properties have been widely studied in the scientific community. However, the effect of central metal ion in the occurrence of their biological properties should be paid more attention. With this aim, novel 2-(hydroxyimino)-1-phenylpropylidene)benzohydrazide (HIPB) Schiff base ligand, and C1/palladium(II), C2/platinum(II), and C3/zinc(II) complexes derived from it were synthesized and characterized. Theoretical studies showed that C2 is more reactive and also has a higher pharmacological affinity than C1 and C3. Experimental investigations were done to compare some biological properties of the complexes. The anticancer assay showed that C1-C3 have the ability to inhibit the growth of HCT116 colon cancer cell lines, but C2 shows a relatively better effect than other. Antioxidant studies using •DPPH (2,2-diphenyl-1-picrylhydrazyl) assay presented the following trend: C2 > C1 > C3 > HIPB. Considering the importance of the antioxidant enzyme catalase in removing reactive oxygen species (ROS), the interaction of C1-C3 with Bovine Liver Catalase (BLC) was evaluated. Kinetic studies showed that C1-C3 can inhibit the catalytic performance of BLC by a similar mechanism, i.e. mixed-type inhibition. Among them, C1 was the strongest inhibitor (Activity inhibition% = 82.2). The C1-C3 quenched the BLC fluorescence emission with dynamic quenching mechanism. The binding affinity to BLC was higher for C1 and C2 than C3. The most important forces in the interaction of C1-C3 with BLC were hydrophobic interactions, which was strongly confirmed by molecular docking data. Tracking the structural changes of catalase showed that BLC undergoes structural changes in the presence of C1 more than C2 and C3.

ZNF8 promotes progression of gastrointestinal cancers via a p53-dependent mechanism

p53 is a critical tumor suppressor, and the disruption of its normal function is often a prerequisite for the development or progression of tumors. Our previous works revealed that multiple members of Krüppel-associated box (KRAB) domain zinc-finger proteins (KZFPs) family regulate p53 transcriptional activity by interacting with it. But the tumor biology functions of these members have not been fully elucidated. Here, the pan-cancer analysis related to gastrointestinal cancers (GICs) revealed that ZNF8, a p53-interacting protein, is an unfavorable prognostic factor for patients with malignancies. ZNF8 interacts with p53 and further depresses its transcriptional activity in colon cancer cells. The knockdown of ZNF8 or the overexpression of ZNF8 inhibits or facilitates the in vitro colony formation, migration, invasion, and angiogenesis of p53+/+ colon cancer HCT116 cells, HepG2 cells and EC109 cells rather than p53−/− colon cancer HCT116 cells and p53-knockout HepG2 cells, respectively. Xenograft experiments conducted in vivo also showed that the knockdown of ZNF8 in p53+/+ but not in p53−/− HCT116 cells curbs the tumor growth and metastasis to lung, leading to an extended life span for tumor-bearing mice. Clinically, two independent immunohistochemistry cohorts of colon cancer and esophageal cancer also indicated that ZNF8 is higher expression in carcinoma tissues than adjacent tissues and this is associated with worse overall survival outcomes in patients without harboring p53 mutation. Together, our results provide insight into the p53-specific tumor oncogenic function of ZNF8. ZNF8 may prove to be a potential target for GICs treatment.

Metabolomic Profiling of Guadua Species and Its Correlation with Antioxidant and Cytotoxic Activities.

Bamboo plants are widely used in Asian traditional medicine for various health issues and exhibit therapeutic potential. Guadua species are renowned bamboos for their high phenolic compound content, including flavonoids and hydroxycinnamic acid derivatives, and possess noteworthy biological properties. Despite this, there is a notable scarcity of research on the chemical and biological aspects of Latin American bamboo leaf extracts (BLEs), especially concerning the Guadua genus. This study aimed to employ a metabolomics approach to integrate the phytochemical and activity profiles of BLEs to identify potential bioactive markers. We determined the metabolic fingerprints of 30 BLEs through HPTLC, HPLC-DAD, UHPLC-QTOF-MS, and 1H-NMR analyses and screened for antioxidant and cytotoxic activities using ABTS, DPPH, and MTT methods. Ultimately, correlation analyses were performed by using chemometric methods and molecular networking. Our findings present a comprehensive chemical characterization, encompassing 40 flavonoids and 9 cinnamic acid derivatives. Notably, most of these compounds have been reported for the first time within the genus, signifying novel discoveries. Additionally, certain compounds identified in other species of the subfamily Bambusoideae provide valuable comparative insights. These compounds demonstrated a significant correlation with antioxidant potential, with values exceeding 100 and 30 μmol of TE/g of extract for ABTS and DPPH, respectively, in the samples. Extracts from G. incana and G. angustifolia exhibited potent cytotoxic effects with IC50 values of 1.23 and 4.73 μg/mL against HCT-116 colon cancer cells, respectively. Notably, glycosylated flavones showed a strong correlation with cytotoxicity. These new findings significantly contribute to our understanding of the chemical composition and biological properties of these often overlooked bamboo species, providing them with important added value and alternative use.

Design, synthesis and antitumor activity of 3,4,5-Trisubstituted Isoxazoles

In this study, 56 isoxazole derivatives were synthesized and their antitumor activity against 9 cancer cells, such as BXPC-3, MiaPaCa-2, PANC-1, MCF-7, HCT-116, HepG2, NCI-H460, A549 and B16 was detected. The results of pharmacological experiments indicated that most compounds exhibit good cytotoxicity against nine cancer cells. Among them, compound 14–3 has an IC50 of 2.4 μM for colon cancer cells HCT-116, which shows the best effect and is better than the broad-spectrum drug 5-Fluorouracil (5-Fu) for the treatment of colon cancer. In addition, 14–3 induces apoptosis and leads to cell cycle arrest in the G2/M phase. Cytotoxicity tests on human normal hepatocytes LO2 also demonstrates that 14–3 is less toxic than 5-Fu with no obvious toxicity. These results suggest that 14–3 is a potential candidate for the development of anti-tumor drugs.

Sexual dimorphism of colorectal cancer in humans and colorectal tumors in a murine model.

In colorectal cancer, men exhibit a higher incidence than women, and there is a disturbance in the levels of sex steroids in serum in patients with this disease. Consistently, in animals, males have greater tumor growth than females in diverse models. Nevertheless, the role of sex steroids is not well established. For that, we analyzed the effect of the principal gonadal sex steroids in both sexes. We determined sex as a statistically risk factor for colorectal cancer with data obtained from GLOBOCAN database. To induce colorectal tumors, we used the gold standard chemical method of azoxymethane and dextran sulphate of sodium. To evaluate the role of sex steroids, we gonadectomized independent males and female animals, reconstituting and substituting them with 17β estradiol and dihydrotestosterone. Finally, we determined, in vitro, the proliferation of a human cell line exposed to 17β estradiol, testosterone, or dihydrotestosterone. Sex, as a risk factor for colorectal cancer, showed a statistically significant susceptibility of men over 50 years old. In vivo, males develop a greater number of tumors and with a larger size than females. In males, orchiectomy prevents tumor growth, whereas in females, ovariectomy promotes the development of neoplasms. DHT acts as a protumoral agent in both sexes. 17β estradiol reduces tumor growth in females but enhances it in males, showing a dimorphic effect. In vitro studies reveal that estradiol decreases the proliferation of the HCT-116 colon cancer cell line, while testosterone boosts proliferation in these cells. Interestingly, dihydrotestosterone does not influence proliferation.

Quantitative structure-activity relationship and ADME prediction studies on series of spirooxindoles derivatives for anti-cancer activity against colon cancer cell line HCT-116

Forty-one derivatives of spirooxindoles, active against HCT-116 colon cancer cells, underwent pharmacophore-based 3D-QSAR analysis to understand their correlation with anti-cancer activity. The study identified a seven-point pharmacophore model (ADHHRRR1) and QSAR models, offering insights for lead optimization and novel analogue design, thus advancing anti-cancer drug discovery. This research underscores the value of molecular modeling in elucidating structure-activity relationships and enhancing drug development efforts.

Targeting colon cancer and normal cells with cold plasma-activated water: Exploring cytotoxic effects and cellular responses

Plasma-activated water (PAW), generated by cold plasma, is emerging as a potential treatment for colon cancer. This study focused on its anticancer effects against HCT-116 colon cancer cells, emphasizing the role of pH and conductivity variations due to plasma–fluid reactions. These changes suggest a chemical transformation in PAW, leading to increased acidity and ion presence. The cytotoxic impact of PAW on HCT-116 cells was analyzed using methods like enzyme-linked immunosorbent assay and microscopic evaluation. PAW exhibited cytotoxicity against HCT-116 cells, but also affected normal colon cells, posing a challenge for selectivity. An 18 h exposure duration was identified as a balance between cancer cell eradication and normal cell preservation. Observed morphological changes indicated apoptotic characteristics in PAW-treated cells, hinting at mechanisms of cancer cell death. PAW-induced reactive oxygen species release mirrored cellular stress, with early apoptotic markers, DNA fragmentation, and increased heat shock proteins (HSPs) signifying complex cellular responses. These findings suggest that PAW can trigger apoptosis and cellular stress pathways cancer cells. However, further studies are necessary for its potential as a cancer therapy.

Red Sea Sponge Extract Callyspongia siphonella and its Metabolites Induced Anticancer Activity in 2D and 3D Culture of Colon Cancer Cells.

Colorectal Cancer (CRC) significantly contributes to global cancer-related mortality and morbidity. Callyspongia siphonella (Callyspongia sp.), a Red Sea sponge, has shown promising activity as an anticancer extract and a source of anticancer-active compounds. This study sought to determine the effects of Callyspongia siphonella and its metabolites on HCT-116 colon cancer cells. Cell viability assays showed that Callyspongia sp. inhibited in a dose-dependent manner, the growth of HCT-116 cell lines with IC50 values of 64.8±17 ug/ml on 2D culture and 141.1±6.8 ug/ml on 3D culture. The purified compounds Sipholenol-A and Sipholenone-A have an IC50 of 48.9±2.2 uM and 47.1±1.2 uM respectively. Following Callyspongia sp. treatment of HCT-116, cell cycle analysis showed arrest at G2/M.flow cytometry analysis showed an increase in total apoptosis due to Callyspongia sp treatment. Moreover, mitochondria membrane potential has been reported to be depolarized due to Callyspongia sp. which is an extra sign of apoptosis. Further investigations are needed to explain the particular underlying mechanisms of Callyspongia sp. extract and its metabolites Sipholenol-A and Sipholenone-A to explore their therapeutic potential in treating colon cancer.

Forming Single-Cell-Derived Colon Cancer Organoid Arrays on a Microfluidic Chip for High Throughput Tumor Heterogeneity Analysis.

Single-cell-derived tumor organoids (STOs) possess a distinct genetic background, making them valuable tools for demonstrating tumor heterogeneity. In order to fulfill the high throughput demands of STO assays, we have developed a microfluidic chip containing 30 000 microwells, which is dedicated to a single cell culture approach for selective expansion and differential induction of cancer stem cells. The microwells are coated with a hydrophilic copolymer to eliminate cell adhesion, and the cell culture is supported by poly(ethylene glycol) (PEG) to establish a nonadhesive culture environment. By utilizing an input cell density of 7 × 103·mL-1, it is possible to construct a 4000 single cell culture system through stochastic cell occupation. We demonstrate that the addition of 15% PEG10000 in the cell culture medium effectively prevents cell loss while facilitating tumor stem cell expansion. As were demonstrated by HCT116, HT29, and SW480 colon cancer cells, the microfluidic approach achieved a STO formation rate of ∼20%, resulting in over 800 STOs generated from a single culture. Comprehensive analysis through histomorphology, immunohistochemistry, drug response evaluation, assessment of cell invasion, and biomarker detection reveals the heterogeneity among individual STOs. Specifically, the smaller STOs exhibited higher invasion and drug resistance capabilities compared with the larger ones. The developed microfluidic approach effectively facilitates STO formation and offers promising prospects for investigating tumor heterogeneity, as well as conducting personalized therapy-focused drug screening.

Sargahydroquinoic Acid from Brown Seaweed Induces Apoptosis in High Glucose Treated-Human Colon Cancer HCT116 Cells

Sargahydroquinoic Acid from Brown Seaweed Induces Apoptosis in High Glucose Treated-Human Colon Cancer HCT116 Cells