Colorectal Carcinoma Cell Lines Research Articles

Anti-inflammatory and Apoptotic Effects of Levisticum Officinale Koch Extracts on HT 29 and Caco-2 Human Colorectal Carcinoma Cell Lines.

Colorectal cancer is among the deadliest cancers in the world. Due to the occurrence of side effects related to current standard therapy, researchers are seeking better alternative treatments. For many years, herbs have been a promising source for discovering therapeutic compounds. Therefore, the primary objective of this research was to examine the distinctive apoptotic and anti-inflammatory properties exhibited by Levisticum officinale Koch (lovage) on HT-29 and Caco-2 cell lines. The maceration method was used to prepare different extracts (ethanol, dichloromethane, petroleum, and residues) from the plant. These extracts were then tested on two colon cancer cell lines - HT-29 and Caco-2 - using the MTT assay to determine the half-maximal inhibitory concentration (IC50) values. In addition, we evaluated the expression levels of several inflammatory genes (IKKb, IKKa, and REIB) using real-time PCR. We also assessed Cox-2 protein expression using western blot analysis. The western blot was also used to analyze apoptosis-related proteins, including Caspase-3, BAX, and Bcl-2. The dichloromethane extract of Levisticum officin (DELO) exhibited a high cytotoxic effect on Caco-2 and HT-29 cell lines, with IC50 values of 106.0±2 μg/mL in HT-29 cells and 175.3±4 μg/mL in Caco-2 cells after 72 hours. None of the lovage extracts showed a significant cytotoxic effect on non-cancerous cells (3T3 cell line). Furthermore, the group treated with DELO showed a lower expression level of inflammatory genes and COX-2 protein compared to the control group. Notably, treatment with DELO resulted in an increase in Caspase-3 protein and BAX/Bcl-2 ratio in both HT-29 and Caco-2 cells. According to this study, DELO has the potential to act as an anti-inflammatory and anti-cancer agent. Further research on the compounds present in DELO and their effect on various signaling pathways could help in the development of new drugs for diseases where inflammation or cells escape from apoptosis play a crucial role.

Abstract LB168: A selective, potent, and orally bioavailable CDK7 inhibitor demonstrates superior anti-cancer activity in colorectal cancer models

Abstract Dysregulated cell cycle and gene expression are pivotal factors in cancer development and the regulators of these processes are attractive targets for cancer treatment. The success of CDK4/6 inhibitors such as palbociclib, ribociclib, and abemaciclib underscores the promising role of targeting cell cycle regulators in anti-cancer therapy. On the other hand, a robust correlation has been established between the overexpression of oncogenes and super-enhancer (SE). The genes associated with SE show increased vulnerability to transcription factor disruption compared to genes with typical enhancers. It has been demonstrated in previous studies that inhibiting transcription factors can effectively decrease the expression of SE-associated oncogenes, including c-MYC, thereby highlighting potential anticancer effects for oncogene-addicted cancers. Currently, agents targeting transcription factors are undergoing clinical trials, offering promising therapeutic options for patients with various types of cancers. CDK7 plays a crucial role in orchestrating cell cycle progression and transcription by phosphorylating CDKs, the C-terminal domain of RNA polymerase II, and hormone receptors like estrogen and androgen receptors. The inhibition of CDK7 has been shown to effectively inhibit tumor growth through various mechanisms, including inducing cell cycle arrest and inhibiting transcription, thereby leading to the death of cancer cells. Notably, ongoing clinical studies have demonstrated that small-molecule CDK7 inhibitors effectively control cancer progression as a stand-alone and in combination with other anti-cancer agents. C-16661, a selective, potent, orally bioavailable, and reversible ATP-competitive inhibitor of CDK7, displayed a single-digit nanomolar IC50 value in both biochemical and cellular assays. It exhibited excellent selectivity in a kinase panel assay, and remarkable potency in a human cancer cell line panel assay. C-16661 treatment resulted in strong inhibition of both its direct and indirect downstream targets including pS5-POLII, pT161-CDK1, c-MYC, and MCL1 in HCT116, a human colorectal carcinoma cell line. While C-16661 exhibited good PK profiles in animals, it demonstrated excellent tumor growth inhibition without any tolerability issues in a colorectal cancer CDX mouse efficacy model. Citation Format: Heuijoon Park, Kyoungwan Seo, Jihyun Yu, Jieun Kim, Min Sung Joo, Sungeun Kim, Joonhyung Lee, Jeonghyeon Lee, Eun-Jung Kim, Joonwoo Nam, Wooseok Han. A selective, potent, and orally bioavailable CDK7 inhibitor demonstrates superior anti-cancer activity in colorectal cancer models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB168.

Evaluation of Antiproliferative and Cytotoxic potential of organic solvent fractions of potential plant sources against Human colorectal carcinoma cell lines

People have been relying and using various medicinal sources for ages throughout the world for alleviation of different ailments. Eighty percent of people worldwide depend on herbal medication for their fundamental medical needs, according to a World Health Organization (WHO) survey. The need for plant-based medications has led to increased studies into the extraction of therapeutic properties from plants. Bioactive compounds are secondary metabolites, sometimes termed nutraceuticals, present in food, providing health benefits.

Structural basis for the preservation of a subset of topologically associating domains in interphase chromosomes upon cohesin depletion

Compartment formation in interphase chromosomes is a result of spatial segregation between euchromatin and heterochromatin on a few megabase pairs (Mbp) scale. On the sub-Mbp scales, topologically associating domains (TADs) appear as interacting domains along the diagonal in the ensemble averaged Hi-C contact map. Hi-C experiments showed that most of the TADs vanish upon deleting cohesin, while the compartment structure is maintained, and perhaps even enhanced. However, closer inspection of the data reveals that a non-negligible fraction of TADs is preserved (P-TADs) after cohesin loss. Imaging experiments show that, at the single-cell level, TAD-like structures are present even without cohesin. To provide a structural basis for these findings, we first used polymer simulations to show that certain TADs with epigenetic switches across their boundaries survive after depletion of loops. More importantly, the three-dimensional structures show that many of the P-TADs have sharp physical boundaries. Informed by the simulations, we analyzed the Hi-C maps (with and without cohesin) in mouse liver and human colorectal carcinoma cell lines, which affirmed that epigenetic switches and physical boundaries (calculated using the predicted 3D structures using the data-driven HIPPS method that uses Hi-C as the input) explain the origin of the P-TADs. Single-cell structures display TAD-like features in the absence of cohesin that are remarkably similar to the findings in imaging experiments. Some P-TADs, with physical boundaries, are relevant to the retention of enhancer–promoter/promoter–promoter interactions. Overall, our study shows that preservation of a subset of TADs upon removing cohesin is a robust phenomenon that is valid across multiple cell lines.

Structural basis for the preservation of a subset of topologically associating domains in interphase chromosomes upon cohesin depletion.

Compartment formation in interphase chromosomes is a result of spatial segregation between euchromatin and heterochromatin on a few megabase pairs (Mbp) scale. On the sub-Mbp scales, topologically associating domains (TADs) appear as interacting domains along the diagonal in the ensemble averaged Hi-C contact map. Hi-C experiments showed that most of the TADs vanish upon deleting cohesin, while the compartment structure is maintained, and perhaps even enhanced. However, closer inspection of the data reveals that a non-negligible fraction of TADs is preserved (P-TADs) after cohesin loss. Imaging experiments show that, at the single-cell level, TAD-like structures are present even without cohesin. To provide a structural basis for these findings, we first used polymer simulations to show that certain TADs with epigenetic switches across their boundaries survive after depletion of loops. More importantly, the three-dimensional structures show that many of the P-TADs have sharp physical boundaries. Informed by the simulations, we analyzed the Hi-C maps (with and without cohesin) in mouse liver and human colorectal carcinoma cell lines, which affirmed that epigenetic switches and physical boundaries (calculated using the predicted 3D structures using the data-driven HIPPS method that uses Hi-C as the input) explain the origin of the P-TADs. Single-cell structures display TAD-like features in the absence of cohesin that are remarkably similar to the findings in imaging experiments. Some P-TADs, with physical boundaries, are relevant to the retention of enhancer-promoter/promoter-promoter interactions. Overall, our study shows that preservation of a subset of TADs upon removing cohesin is a robust phenomenon that is valid across multiple cell lines.

Unveiling biological activities of biosynthesized starch/silver-selenium nanocomposite using Cladosporium cladosporioides CBS 174.62

Background and objectivesMicrobial cells capability to tolerate the effect of various antimicrobial classes represent a major worldwide health concern. The flexible and multi-components nanocomposites have enhanced physicochemical characters with several improved properties. Thus, different biological activities of biosynthesized starch/silver-selenium nanocomposite (St/Ag-Se NC) were assessed.MethodologyThe St/Ag-Se NC was biosynthesized using Cladosporium cladosporioides CBS 174.62 (C. cladosporioides) strain. The shape and average particle size were investigated using scanning electron microscope (SEM) and high-resolution transmission electron microscope (HR-TEM), respectively. On the other hand, the St/Ag-Se NC effect on two cancer cell lines and red blood cells (RBCs) was evaluated and its hydrogen peroxide (H2O2) scavenging effect was assessed. Moreover, its effects on various microbial species in both planktonic and biofilm growth forms were examined.ResultsThe St/Ag-Se NC was successfully biosynthesized with oval and spherical shape and a mean particle diameter of 67.87 nm as confirmed by the HR-TEM analysis. St/Ag-Se NC showed promising anticancer activity toward human colorectal carcinoma (HCT-116) and human breast cancer (MCF-7) cell lines where IC50 were 21.37 and 19.98 µg/ml, respectively. Similarly, little effect on RBCs was observed with low nanocomposite concentration. As well, the highest nanocomposite H2O2 scavenging activity (42.84%) was recorded at a concentration of 2 mg/ml. Additionally, Staphylococcus epidermidis (S. epidermidis) ATCC 12,228 and Candida albicans (C. albicans) ATCC 10,231 were the highly affected bacterial and fungal strains with minimum inhibitory concentrations (MICs) of 18.75 and 50 µg/ml, respectively. Moreover, the noticeable effect of St/Ag-Se NC on microbial biofilm was concentration dependent. A high biofilm suppression percentage, 87.5% and 68.05%, were recorded with S. epidermidis and Staphylococcus aureus (S. aureus) when exposed to 1 mg/ml and 0.5 mg/ml, respectively.ConclusionThe biosynthesized St/Ag-Se NC showed excellent antioxidant activity, haemocompatibility, and anti-proliferative effect at low concentrations. Also, it exhibited promising antimicrobial and antibiofilm activities.

Anticancer Activities of Zinc Oxide Nanoparticles (ZnONPs) Synthesized from Mentha longifolia L. Leaf Extract

Recently, there has been a remarkable increase in cancer and cancer-related deaths. In this study, the impacts of zinc oxide nanoparticles (ZnONPs) produced from the aqueous leaf extract of Mentha longifolia L. (ML) on ovary adenocarcinoma (OVCAR-3), colorectal carcinoma (HCT-116), and healthy retinal pigment epithelial cell (RPE-1) lines were investigated. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test was performed to discover its antiproliferative properties. As a result of the application of ML-ZnONPs on RPE-1, OVCAR-3, and HCT-116 cell lines at doses (µg/mL) of 250, 500, and 1000 for 24 hours, the viability rates (%) in the cell lines were 18.73-30.56, 21.98-28.76, and 27.27-40.93, respectively. In the 48-hour application, the viability rates (%) of the same cells were between 29.51-46.83, 32.49-40.81, and 46.82-44.37, respectively. The MTT test revealed that ML-ZnONPs strongly suppressed the growth of RPE-1, OVCAR-3, and HCT-116 cell lines. The test showed that the effect of dose increase and time on the viability of both cancer cell lines was negative.

Selective Synergy of Rapamycin Combined With Methioninase on Cancer Cells Compared to Normal Cells.

Rapamycin and recombinant methioninase (rMETase) have both shown efficacy to target cancer cells. Rapamycin prevents cancer-cell growth by inhibition of the mTOR protein kinase. rMETase, by degrading methionine, targets the methionine addiction of cancer and has been shown to improve the efficacy of chemotherapy drugs. In the present study, we aimed to determine if a synergy exists between rapamycin and rMETase when used in combination against a colorectal-carcinoma cell line, compared to normal fibroblasts, in vitro. The half-maximal inhibitory concentrations (IC50) of rapamycin alone and rMETase alone against the HCT-116 human colorectal-cancer cell line and Hs-27 human fibroblasts were determined using the CCK-8 Cell Viability Assay. After calculating the IC50 of each drug, we determined the efficacy of rapamycin and rMETase combined on both HCT-116 and Hs-27. Hs-27 normal fibroblasts were more sensitive to rapamycin than HCT-116 colon-cancer cells (IC50=0.37 nM and IC50=1.38 nM, respectively). HCT-116 cells were more sensitive to rMETase than Hs-27 cells (IC50 0.39 U/ml and IC50 0.96 U/ml, respectively). The treatment of Hs-27 cells with the combination of rapamycin (IC50=0.37 nM) and rMETase (IC50=0.96 U/ml) showed no significant difference in their effect on Hs-27 cell viability compared to the two drugs being used separately. However, the treatment of HCT-116 cells with the combination of rapamycin (IC50=1.38 nM) and rMETase (IC50=0.39 U/ml) was able to decrease cancer-cell viability significantly more than either single-drug treatment. Rapamycin and rMETase, when used in combination against colorectal-cancer cells, but not normal fibroblasts, in vitro, have a cancer-specific synergistic effect, suggesting that the combination of these drugs can be used as an effective, targeted cancer therapy.

Expression of Mitochondrial Long Non-Coding RNAs, MDL1 and MDL1AS, Are Good Prognostic and/or Diagnostic Biomarkers for Several Cancers, Including Colorectal Cancer.

Non-coding RNAs provide new opportunities to identify biomarkers that properly classify cancer patients. Here, we study the biomarker status of the mitochondrial long non-coding RNAs, MDL1 and MDL1AS. Expression of these genes was studied in public transcriptomic databases. In addition, a cohort of 69 locally advanced rectal cancer (LARC) patients with a follow-up of more than 5 years was used to determine the prognostic value of these markers. Furthermore, cell lines of colorectal (HCT116) and breast (MDA-MB-231) carcinoma were employed to study the effects of downregulating MDL1AS in vitro. Expression of MDL1AS (but not MDL1) was significantly different in tumor cells than in the surrounding tissue in a tumor-type-specific context. Both MDL1 and MDL1AS were accurate biomarkers for the 5-year survival of LARC patients (p = 0.040 and p = 0.007, respectively) with promising areas under the curve in the ROC analyses (0.820 and 0.930, respectively). MDL1AS downregulation reduced mitochondrial respiration in both cell lines. Furthermore, this downregulation produced a decrease in growth and migration on colorectal cells, but the reverse effects on breast cancer cells. In summary, MDL1 and MDL1AS can be used as reliable prognostic biomarkers of LARC, and MDL1AS expression provides relevant information on the diagnosis of different cancers.

A37 POSTBIOTIC BACTERIAL MEMBRANE VESICLES FROM B. SUBTILIS DELIVER NOD2 LIGANDS AND PROMOTE IN VITRO WOUND RE-EPITHLIALIZATION THROUGH RIPK2 SIGNALING

Abstract Background Bacterial membrane vesicles (MVs) are bilipid nanoparticles secreted as a conserved method of intercellular communication. MVs from Gram-positive bacteria carry diverse bacterial products and represent a unique opportunity in the growing field of postbiotics. This native machinery for mediating microbe-host interactions makes MVs a promising tool for intestinal drug delivery in the context of Crohn’s disease (CD). CD is a chronic inflammatory bowel disease characterized by relapsing inflammation of the digestive tract. CD is associated with (1) loss of function mutations in the host gene encoding the nuclear-binding oligomerization domain 2 (NOD2) pattern recognition receptor and (2) a gut microbiome with reduced capabilities to generate NOD2 stimulating muropeptides from peptidoglycan. Nod2-/- mice have decreased barrier integrity and impaired epithelial restitution upon challenge. However, there are currently no therapeutic strategies targeting NOD2 for CD management. Aims The aims of this study were to (1) leverage MVs as a biological system for NOD2 ligand delivery to intestinal epithelial cells and (2) determine the roll of probiotic MVs and bacterial ligand synergy in wound re-epithelialization. Methods MVs were purified from B. subtilis through filtration and ultracentrifugation. MVs were characterized through nanoparticle tracking analysis using NanoSight300. WT and NOD2-/- HCT116 cells, a human colorectal carcinoma cell line, were utilized for IL-8 secretion quantification by ELISA, gene expression by qPCR, and in vitro scratch wound assay by Incucyte quantification. Results Here, we demonstrate for the first time that MVs from B. subtilis deliver NOD2 ligands in vitro. Treatment with isolated MVs induces IL8 secretion and CXCL1 expression in a NOD2-dependent manner. Furthermore, MVs promote re-epithelialization after in vitro scratch wounding. This effect was completely blunted by the addition of an inhibitor for RIPK2, a downstream transducer required for NOD2 signaling. Through bacterial pathogen-associated molecular patterns (PAMPs) screening, we found that wound re-epithelialization is promoted by PAMP synergy but dependent on RIPK2 signaling. Conclusions This work suggests that delivery of muropeptides by probiotic-derived MVs is a promising strategy to promote epithelial regeneration during injury through targeting NOD2. Future directions will work towards validating these findings in human ileal primary cells and murine models of intestinal injury. Funding Agencies CIHRMedicine by Design

Antiproliferative and Antitelomerase Effects of Silymarin on Human Colorectal and Hepatocellular Carcinoma Cells.

Silymarin, a widely-used hepatoprotective agent, has shown antitumor properties in both in vitro and animal studies. Currently, there is limited knowledge regarding silymarin's antitelomerase effects on human colorectal cancer and hepatocyte carcinoma cells. In this study, we investigated the antiproliferative and antitelomerase effects of silymarin on four human colorectal cancer and HepG2 hepatocyte carcinoma cell lines. The cell viability and telomerase activity were assessed using MTT and the telomerase repeat amplification protocol assay, respectively. We also investigated the effects of silymarin on the expression of human telomerase reverse transcriptase and its promoter methylation in HepG2 cells by real-time RT-PCR and methylation-specific PCR, respectively. Silymarin treatment inhibited cell proliferation and telomerase activity in all cancer cells. After 24 h of treatment, silymarin exhibited IC50 values ranging from 19 - 56.3 µg/mL against these cancer cells. A 30-min treatment with silymarin at the IC50 concentration effectively inhibited telomerase activity in cell-free extracts of both colorectal cancer and hepatocyte carcinoma cells. Treatment of HepG2 cells with 10 and 30 µg/mL of silymarin for 48 h resulted in a decrease in human telomerase reverse transcriptase expression to 75 and 35% of the level observed in the untreated control (p < 0.01), respectively. Treatment with silymarin (10, 30, and 60 µg/mL) for 48 h did not affect human telomerase reverse transcriptase promoter methylation in HepG2 cells. In conclusion, our findings suggest that silymarin inhibits cancer cell growth by directly inhibiting telomerase activity and downregulating its human telomerase reverse transcriptase catalytic subunit. However, silymarin did not affect human telomerase reverse transcriptase promoter methylation at the concentrations of 10 - 60 µg/mL used in this study.

Phytochemical Contents and Anti-Colon Cancer Activities of Macadamia integrifolia Maiden &amp; Betche Leaves and Husks Extracts

The macadamia (Macadamia integrifolia Maiden &amp; Betche, Proteaceae Family) leaves and husks were the by-products from macadamia nut production which have few reports on the phytochemicals and pharmacological activities. This study aimed to investigate the possibility of using these by-products in drug discovery by examining their phytochemicals contents, as well as their anti-colon cancer activities. Dry macadamia leaves and husks were extracted and fractionated to obtain the crude extracts (EtOH), dichloromethane (DCM), ethyl acetate (EtOAc) and water fractions. Colorimetric methods were used to determine the total amounts of phytochemicals. The anti-colon cancer activities of the extracts against human colorectal carcinoma (HCT116) cell line were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The results showed that each gram of the samples contained total phenolics between 2.9-12.9 mg equivalent to gallic acid, total flavonoids 30.1-89.7 mg equivalent to rutin, and total triterpenoids 228.4-640.9 mg equivalent to ursolic acid. The DCM and EtOAc fractions of the husks exhibited the highest anti-colon cancer activity, with half maximal inhibitory concentrations (IC50) of 39.1 ± 4.3 and 56.9 ± 15.0 µg/mL, respectively. The EtOAc and water fractions of the leaves had IC50 values of 130.9 ± 17.4 and 127.8 ± 7.9 µg/mL, respectively. The IC50 values of other samples were greater than 200 µg/mL, while the IC50 of sorafenib, a positive control, was 5.95 ± 1.26 µg/mL. In conclusion, the leaves and husks of macadamia contained high amounts of several phytochemicals and exhibited anti-colon cancer activity against HCT116. These findings suggested that the extracts from these wastes may have potential in the discovery of anticancer agent research. Keywords: Macadamia integrifolia, Leaf, Husk, Colon cancer, Phytochemical

Anti-MRSA and cytotoxic activities of different solvent extracts from Artemisia herba-alba grown in Shubak, Jordan.

Globally, resistance to antimicrobial drugs is a major hazard to public health. Infections that were once easily treatable with antibiotics are becoming harder to control, leading to prolonged illnesses, increased mortality rates, and higher healthcare costs. This study intended to assess the antimicrobial, specifically the anti-Methicillin resistant Staphylococcus aureus (MRSA), and anticancer properties of different extracts obtained from A. herba-alba (AHA). The antibacterial tests of AHA were performed on two Gram-negative bacterial strains (Escherichia coli and Klebsiella pneumonia), two Gram-positive bacterial strains (Methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus aureus). Initial screening for antibacterial activities was conducted using the well diffusion technique. Subsequently, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined through the broth-dilution assay. The anticancer test was carried out in vitro on a human colorectal carcinoma cell line (HCT-116) using MTT assay. Among all extracts, n-hexane extract of AHA was the most effective against S. aureus with the highest inhibition zone (24.67 mm ± 0.58) compared to standard antibiotic (erythromycin, 24.00 mm) followed by the methanolic extract against MRSA (24.00 mm ± 1.73). The methanol extract of AHA showed the highest antibacterial activity against MRSA. The results of MIC and MBC of the AHA methanol extract against MRSA were 1.17 ± 1.09 and 9.375 ± 0.0 mg/ml, respectively, demonstrating therapeutically significant antibacterial activity. Ethyl acetate extract has no antibacterial activity against E. coli and K. pneumonia. The findings indicated that the methanol extract of AHA exhibited the highest efficacy against the colorectal carcinoma cell line (HCT-116), with an IC50 value of 126.61 ± 13.35 μg/ml. These findings suggest that the methanol extract of AHA could be considered as a potential agent to serve as a source of antibacterial and anticancer compounds.

Crude Polysaccharides from Mushrooms Elicit an Anti-Allergic Effect Against Type 1 Allergy In Vitro.

Type 1 allergic disease is a global challenge, hence the search for alternative therapies. Mushrooms have several medicinal and health benefits. However, scant data exist on the anti-allergic properties of polysaccharides from fruiting bodies (FB) and mycelia of mushrooms. We used an in vitro co-culture system comprising Caco-2 cells (intestinal epithelial colorectal carcinoma cell line) and RBL-2H3 cells (cell line from rat basophilic leukemia cells). Reduction in degranulation of mast cells indicated anti-allergy properties. The inhibitory effect of crude polysaccharides from different mushroom FB and mycelia on β-hexosaminidase release from RBL-2H3 cells was measured. Results showed that crude polysaccharides from the FB of Inonotus obliquus exhibited a significant inhibitory effect on β-hexosaminidase release and lowered it by 16%. Polysaccharides from the FB of Lentinus squarrosulus, and Pleurotus ostreatus did not exhibit a significant reduction in β-hexosaminidase. However, crude polysaccharides from their mycelia had a significant inhibitory effect, resulting in up to a 23% reduction in β-hexosaminidase activity. Among fungi showing degranulation properties, crude polysaccharides from their mycelia showed more potent action against degranulation than their corresponding FB. Polysaccharides extracted from FB and or mycelia, of selected mushrooms, possess anti-allergic properties that could be harnessed for use in alternative allergy therapies.

Retrospective Study of the Use of Heparins in Pregnant Women and in vitro Testing on the HCT 116 Colorectal Carcinoma Cell Line.

Pregnant women manifest an increased risk of developing coagulation disorders. Unfractionated heparin (HEP) and low-molecular-weight heparin (LMWHep) are considered as selective medication in the case of pregnancy which needs anticoagulant treatment. In addition to anticoagulant properties, HEP and its derivatives manifest other properties including anti-cancer potential. According to Globocan's latest data, colorectal cancer (CRC) is the second most encountered form of malignancy in the case of women, manifesting some special particularities, as confusion of symptoms from cancer with symptoms encountered normally in pregnant women (such as constipation or rectal bleeding), delayed diagnosis because of limitations imposed both for the fetus and for the mother, and the need for special treatment. The aim of the present work is to follow the incidence and safety of consumption of HEP and LMWHep in the case of pregnant women and to analyze their potential on the HCT 116 colorectal carcinoma cells. Analyzing the consumption of heparins in case of pregnant women hospitalized from 01.01.2022 to 31.12.2022 at the Pius Brînzeu" Emergency Clinical Hospital from Timisoara, Obstetrics and Gynecology Clinic I, it was observed that 44,6% of the patients were administered the following medication and no administration risks were observed. When tested on HCT 116 cells, heparins manifested a significant anti-migratory effect (with wound healing rates of 2,6%, when tested with HEP 100 UI concentration and 14.52% wound healing rates in case of fraxiparine 100 UI). In addition, different signs of apoptosis were observed, suggesting the pro-apoptotic potential of the tested substances. Heparins remain the preferred medication to be administered to pregnant women with the potential for coagulation disorders, showing a high safety profile. Testing on the cancerous line of colorectal carcinoma highlights important properties that stimulate future studies, to establish the anti-tumor potential and the exact mechanism of action.

EBP50 Depletion and Nuclear β-Catenin Accumulation Engender Aggressive Behavior of Colorectal Carcinoma through Induction of Tumor Budding.

Ezin-radixin-moesin-binding phosphoprotein 50 (EBP50) is a scaffold protein that interacts with several partner molecules including β-catenin. Here, we examined the crosstalk between EBP50 and nuclear catenin during colorectal carcinoma (CRC) progression. In clinical samples, there were no correlations between the subcellular location of EBP50 and any clinicopathological factors. However, EBP50 expression was significantly lower specifically in the outer areas of tumor lesions, in regions where tumor budding (BD) was observed. Low EBP50 expression was also significantly associated with several unfavorable prognostic factors, suggesting that EBP50 depletion rather than its overexpression or subcellular distribution plays an important role in CRC progression. In CRC cell lines, knockout of EBP50 induced epithelial-mesenchymal transition (EMT)-like features, decreased proliferation, accelerated migration capability, and stabilized nuclear β-catenin due to disruption of the interaction between EBP50 and β-catenin at the plasma membrane. In addition, Slug expression was significantly higher in outer lesions, particularly in BD areas, and was positively correlated with nuclear β-catenin status, consistent with β-catenin-driven transactivation of the Slug promoter. Together, our data suggest that EBP50 depletion releases β-catenin from the plasma membrane in outer tumor lesions, allowing β-catenin to accumulate and translocate to the nucleus, where it transactivates the Slug gene to promote EMT. This in turn triggers tumor budding and contributes to the progression of CRC to a more aggressive phase.

Tumor-immune cell interaction and senescence-associated molecules in colorectal carcinoma

Cellular senescence permanently arrests the cell cycle of premalignant cells following protumorigenic stimuli, counteracting tumor progression. Senescence induction leads to phenotypic and metabolic changes and alters the interaction with the cells' microenvironment. This mediates tumor immunosurveillance but bears promalignant potential and may contribute to disease progression. Our study aims to investigate the prognostic potential of senescence markers in colorectal carcinoma (CRC) and to understand the interaction of senescent tumor cells and immune cells. Immunohistochemical markers were studied on atissue microarray (TMA) containing tumor tissue of n = 598 CRC patients and were evaluated using digital image analysis. Results were correlated with disease-specific survival (DSS) and progression-free survival (PFS). Consecutive TMA sections were stained for senescence markers and immune cell markers to analyze the spatial relation of those cell populations. Senescence was induced in CRC cell lines in vitro and co-cultures with various immune cell lines were established to study the interactions. Expression of different senescent-associated markers is associated with increased or decreased DSS and PFS. Close proximity of p21+ senescent tumor cells and CD8+ immune cells correlates with increased DSS and PFS. In vitro, senescent cells were dose-dependently eliminated by immune cells, which is facilitated via direct cell-cell contact and induction of apoptosis. Depicting the initiation of this important anti-tumor mechanism, markers of cellular senescence are of significant prognostic relevance in CRC. Moreover, our results show the pleiotropic effect of senescence in vivo. Absence as well as exceeding expression of senescence markers are associated with anegative prognosis in CRC. The impact of cellular senescence depends on the tumor microenvironment and the immunosurveillance of senescent cells. Proximity analyses of senescent cells and tumor-infiltrating immune cells have significant prognostic relevance and reflect this.

Anticancer Potential of Farnesol Against Human Osteosarcoma Saos-2 Cells and Human Colorectal Carcinoma HCT-116 Cells.

Increased colorectal carcinoma (CRC) and osteosarcomaprevalence, low survival rate, poor prognosis, and the limitations of existing anticancer therapies like side effects of drugs, non-specificity, short half-life, etc., pose a need for novel anticancer drugs.Farnesol, an organic sesquiterpene compound, found in the essential oils of various plants has been shown to possess antioxidant, anti-inflammatory, and anticancer properties. However, the anticancer effect of farnesol againstCRCand osteosarcoma has not yet been adequately elucidated. The aim of the study was to analyze the anticancer effects of farnesol against human osteosarcoma and CRC cell lines. Human osteosarcoma (Saos-2) and colorectal carcinoma (HCT-116) cell lines were procured and cultured at37oC and 5% CO2. The cells were treated with 10, 20, 40, 60, 80, and 100 µM/ml and 20, 40, 60, 80, 100, and 120 µM/ml of farnesol for 24 hours, respectively.3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromideassay was performed to assess the cytotoxicity of farnesol on Saos-2 and HCT-116 cells. Acridine orange/ethidium bromidestaining was carried out to analyze apoptosis.4',6-diamidino-2-phenylindolestaining was done to observe the nuclear changes.Dichloro-dihydro-fluorescein diacetatestaining was performed to assess the farnesol-induced reactive oxygen species (ROS)-mediated cell death. Farnesol reduced the viability and proliferation of Saos-2 and HCT-116 cells in a dose-dependent manner. Farnesol was able to alter the cellular and nuclear morphology of Saos-2 and HCT-116 cells, promoting cell death. Farnesol-induced apoptosis inhuman osteosarcoma and colorectal carcinoma cell lines. Early apoptosis was observed in farnesol-treated HCT-116 cells. Additionally, ROS-mediated apoptotic cell death was reported in Saos-2 cells. Farnesol has the potential to induce cytotoxicity against human osteosarcoma and CRC cell lines.

New cytotoxic dammarane type saponins from Ziziphus spina-christi

Cancer is the world's second-leading cause of death. Drug development efforts frequently focus on medicinal plants since they are a valuable source of anticancer medications. A phytochemical investigation of the edible Ziziphus spina-christi (F. Rhamnaceae) leaf extract afforded two new dammarane type saponins identified as christinin E and F (1, 2), along with the known compound christinin A (3). Different cancer cell lines, such as lung cancer (A549), glioblastoma (U87), breast cancer (MDA-MB-231), and colorectal carcinoma (CT-26) cell lines, were used to investigate the extracted compounds' cytotoxic properties. Our findings showed significant effects on all the tested cell lines at varying concentrations (1, 5, 10, and 20 µg/mL). The three compounds exhibited potent activity at low concentrations (< 10 μg/mL), as evidenced by their low IC50 values. To further investigate the complex relationships between these identified cancer-relevant biological targets and to identify critical targets in the pathogenesis of the disease, we turned to network pharmacology and in silico-based investigations. Following this, in silico-based analysis (e.g., inverse docking, ΔG calculation, and molecular dynamics simulation) was performed on the structures of the isolated compounds to identify additional potential targets for these compounds and their likely interactions with various signalling pathways relevant to this disease. Based on our findings, Z. spina-christi's compounds showed promise as potential anti-cancer therapeutic leads in the future.

SOX8 promotes tumor growth and metastasis through FZD6-dependent Wnt/β-catenin signaling in colorectal carcinoma

SOX8 plays an important role in several physiological processes. Its expression is negatively associated with overall survival in patients with colorectal carcinoma (CRC), suggesting SOX8 is a potential prognostic factor for this disease. However, the role of SOX8 in CRC remains largely unknown. In this study, our data showed that SOX8 expression was upregulated in CRC cell lines and tumor tissues. Stable knockdown of SOX8 in CRC cell lines dramatically reduced cell proliferation, migration, and invasion. Furthermore, the knockdown of SOX8 decreased the phospho-GSK3β level and suppressed Frizzled-6 (FZD6) transcription; restoration of FZD6 expression partially abolished the effect of SOX8 on Wnt/β-catenin signaling and promote CRC cell proliferation. In conclusion, our findings suggested that SOX8 served as an oncogene in CRC through the activation of FZD6-dependent Wnt/β-catenin signaling.