Colon Cancer Cell Lines Research Articles

Bee (Apis mellifera L. 1758) wax restores adipogenesis and lipid accumulation of 3T3-L1 cells in cancer-associated cachexia condition.

Cachexia is associated with various diseases, such as heart disease, infectious disease, and cancer. In particular, cancer-associated cachexia (CAC) accounts for more than 20% of mortality in cancer patients worldwide. Adipose tissue in CAC is characterized by adipocyte atrophy, mainly due to excessively increased lipolysis and impairment of adipogenesis. CAC is well known for the loss of skeletal muscle mass and/or fat mass. CAC induces severe metabolic alterations, including protein, lipid, and carbohydrate metabolism. The objectives of this study were to evaluate the effects of bee wax (Apis mellifera L. 1758) (BW) extract on adipogenesis, lipolysis, and mitochondrial oxygen consumption through white adipocytes, 3T3-L1. To achieve this study, cancer-associated cachexia condition was established by incubation of 3T3-L1 with colon cancer cell line CT26 cultured media. BW extract recovered the reduced adipogenesis under cachectic conditions in CT26 media. Treatment of BW showed increasing lipid accumulation as well as adipogenic gene expression and its target gene during adipogenesis. The administration of BW to adipocytes could decrease lipolysis. Also, BW could significantly downregulated the mitochondrial fatty acid oxidation-related genes, oxygen consumption rate, and extracellular acidification rate. Our results suggest that BW could improve metabolic disorders such as CAC through the activation of adipogenesis and inhibition of lipolysis in adipocytes, although we need further validation invivo CAC model to check the effects of BW extract. Therefore, BW extract supplements could be useful as an alternative medicine to reverse energy imbalances.

In Vitro Inhibition of Colorectal Cancer Gene Targets by Withania somnifera L. Methanolic Extracts: A Focus on Specific Genome Regulation.

An approach that shows promise for quickening the evolution of innovative anticancer drugs is the assessment of natural biomass sources. Our study sought to assess the effect of W. somnifera L. (WS) methanolic root and stem extracts on the expression of five targeted genes (cyclooxygenase-2, caspase-9, 5-Lipoxygenase, B-cell lymphoma-extra-large, and B-cell lymphoma 2) in colon cancer cell lines (Caco-2 cell lines). Plant extracts were prepared for bioassay by dissolving them in dimethyl sulfoxide. Caco-2 cell lines were exposed to various concentrations of plant extracts, followed by RNA extraction for analysis. By explicitly relating phytoconstituents of WS to the dose-dependent overexpression of caspase-9 genes and the inhibition of cyclooxygenase-2, 5-Lipoxygenase, B-cell lymphoma-extra-large, and B-cell lymphoma 2 genes, our novel findings characterize WS as a promising natural inhibitor of colorectal cancer (CRC) growth. Nonetheless, we recommend additional in vitro research to verify the current findings. With significant clinical benefits hypothesized, we offer WS methanolic root and stem extracts as potential organic antagonists for colorectal carcinogenesis and suggest further in vivo and clinical investigations, following successful in vitro trials. We recommend more investigation into the specific phytoconstituents in WS that contribute to the regulatory mechanisms that inhibit the growth of colon cancer cells.

Anticancer Activity of Rice Callus Suspension Cultures from Aromatic Varieties and Metabolites Regulated in Treated Cancer Cell Lines

Tissue culture techniques were used to produce large amounts of bioactive compounds with medicinal potential, overcoming space and time constraints for cancer prevention. Rice callus suspension cultures (RCSC) and seed extracts prepared from aromatic rice varieties were used to evaluate the cytotoxic impact on human colon and lung cancer cell lines, as well as a normal control cell line, using Taxol as a positive control. RCSC and seed extracts from two Indian aromatic rice varieties were applied at different concentrations to treat the cancer cell lines and normal lung fibroblasts over varying time intervals. Apoptosis was assessed in 1:5 dilutions of the A549 and HT-29 cell lines treated with RCSC for 72 h, using propidium iodide staining and flow cytometry. RCSC showed a more potent cytotoxic effect than seed extracts with minimal effect on the normal cell line, in contrast to Taxol. Confocal microscopy and flow cytometry further confirmed the apoptotic effect of RCSC. Gas chromatography-mass spectrometry-based metabolic profiling identified metabolites involved in cytotoxicity and highlighted altered pathways. RCSC is proposed as an alternative source for the development of novel anticancer drugs with reduced side effects.

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

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

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

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

LncRNA EBLN3P Accelerates Cell Proliferation and Invasion of Colon Cancer through Modulating the miR-519d-3p/ZFP91 Axis.

Purpose: The study aims to explore the roles and underlying mechanisms of long noncoding RNAs endogenous bornavirus-like nucleoprotein (lncRNA EBLN3P) in colon cancer, emphasizing the potential impact of these insights on advancing colon cancer treatment strategies. By shedding light on lncRNA EBLN3P's involvement, this research could contribute to the development of novel therapeutic approaches, enhancing the efficacy of interventions for colon cancer patients. Methods: We employed quantitative reverse transcription polymerase chain reaction to assess the levels of lncRNA EBLN3P, zinc finger protein (ZFP91), and miR-519d-3p, alongside CCK-8 and EdU assays for cell proliferation, flow cytometry for apoptosis, and Transwell and wound healing assays for migration and invasion. The in vivo function of lncRNA EBLN3P was investigated through a xenograft model, and protein levels were evaluated via Western blot analysis. Results: LncRNA EBLN3P was found to be upregulated in colon cancer tissues and cells, promoting cell proliferation and metastasis while inhibiting apoptosis. Downregulation of lncRNA EBLN3P reduced tumor size, volume, and weight in a mouse model. MiR-519d-3p, which negatively interacts with lncRNA EBLN3P, was found to be downregulated in colon cancer tissues and cell lines. Its upregulation hindered cancer cell proliferation and metastasis while enhancing apoptosis. ZFP91, a binding partner of miR-519d-3p, was upregulated in colon cancer and inversely related to miR-519d-3p levels. Rescue experiments indicated that the effects of lncRNA EBLN3P silencing could be reversed by miR-519d-3p suppression, but were mitigated by ZFP91 downregulation. Conclusion: LncRNA EBLN3P facilitates colon cancer progression via the miR-519d-3p/ZFP91 axis, presenting a novel understanding of lncRNA EBLN3P's role and offering potential therapeutic insights for colon cancer treatment. This study fills a critical gap by linking lncRNA EBLN3P with the miR-519d-3p/ZFP91 axis in the context of colon cancer, thereby broadening our understanding of the molecular mechanisms underlying colon cancer progression.

Exploring the impact of MiR-92a-3p on FOLFOX chemoresistance biomarker genes in colon cancer cell lines.

Introduction: One of the primary obstacles faced by individuals with advanced colorectal cancer (CRC) is the potential development of acquired chemoresistance as the disease advances. Studies have indicated a direct association between elevated levels of miR-92a-3p and the progression, metastasis, and chemoresistance observed in CRC. We proposed that miR-92a-3p impairs FOLFOX (fluorouracil/oxaliplatin) chemotherapy response by upregulating the expression of chemoresistance biomarker genes through the activation of β-catenin and epithelial-mesenchymal transition (EMT). These FOLFOX biomarker genes include the pyrimidine biosynthesis pathway genes dihydropyrimidine dehydrogenase (DPYD), thymidylate synthase (TYMS), methylenetetrahydrofolate reductase (MTHFR), and the genes encoding the DNA repair complexes subunits ERCC1 and ERCC2, and XRCC1. Methods: To assess this, we transfected SW480 and SW620 colon cancer cell lines with miR-92a-3p mimics and then quantified the expression of DPYD, TYMS, MTHFR, ERCC1, ERCC2, and XRCC1, the expression of EMT markers and transcription factors, and activation of β-catenin. Results and discussion: Our results reveal that miR-92a-3p does not affect the expression of DPYD, TYMS, MTHFR, and ERCC1. Furthermore, even though miR-92a-3p affects ERCC2, XRCC1, E-cadherin, and β-catenin mRNA levels, it has no influence on their protein expression. Conclusion: We found that miR-92a-3p does not upregulate the expression of proteins of DNA-repair pathways and other genes involved in FOLFOX chemotherapy resistance.

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 LB160: The mouse mammary tumor virus mediates the antineoplastic action of decitabine

Abstract The purpose of this study is to clarify the role of the mouse mammary tumor virus (MMTV) in cancer growth and in mediating the antineoplastic effect of decitabine, a DNA demethylating agent. In vitro studies have shown that decitabine inhibits cancer cell proliferation by stimulating endogenous retroviruses which in turn activates the production of cytostatic interferon beta (IFN-β). We investigated the potential involvement of endogenous MMTV and the mouse IFN-β in the pharmacodynamics of decitabine using stable knockdown cell lines and MMTV hyper-infection. For the first time, we show that knockdown of MMTV or IFN-β in a mouse mammary cancer cell line (4T1) both rendered the cancer cells more resistant to decitabine in mice bearing 4T1-derived mammary cancer. Mechanistically, decitabine enhanced MMTV expression on the RNA level in both control tumors and tumors with MMTV knockdown, although the degree of enhancement was limited in knockdown tumors. Conversely, infection of a mouse colon cancer cell line (MC38) with MMTV released from 4T1 cells rendered the colon cancer cells more sensitive to decitabine. These data confirm the viral mimicry hypothesis as the mechanism of DNA demethylating agents. Practically, our research suggests the possibility of combining the use of an exogenous virus and a DNA demethylating agent as a new approach to cancer treatment. Citation Format: Sarah Light, Maziar Montazer, Cade Wycoff, Richard Tuttle, Andrew Brola, William Wycoff, Ryan Johnson, Yingguang Liu. The mouse mammary tumor virus mediates the antineoplastic action of decitabine [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 LB160.

Abstract LB322: ARC2-001, a CAF-targeting drug exhibits strong anticancer effect in in vivo setting

Abstract For more effective tumor growth suppression, a combinatorial treatment of CAF-targeting agent and other well-validated therapies such as chemotherapy, radiotherapy, and immune checkpoint therapy should be considered. Thus, we hypothesized that co-administration of the CAF-targeting drug, ARC2-001 (the in vitro-approved CAF-targeting candidate) and clinically practiced anticancer drugs could be useful strategy for treating under clinical settings. However, it is essentially important to evaluate this using well-established pre-clinical models. We thoroughly verified the anticancer effect of ARC-001 using two in vivo model systems. ( 2nd- in vivo systems for CAF targeted drugs Research). 1. (Subcutaneous model) Immune-deficient mice (NOD/SCID): in which human colon cancer cell line HCT116 was co-injected with human CAF (CAFs were obtained from stage IV CRC patients) followed by administration of ARC2-001 alone or in combination with 5-fluorouracil (5FU). 2. (Subcutaneous and Orthotopic model) Wild-type mice (C57BL/6): in which mouse colon cancer cells was co-injected with mouse colon CAFs (both cancer cells and CAFs were obtained from chemically-induced colorectal cancer model with rich desmoplastic stroma) followed by administration of ARC2-001 alone or in combination with aPD-1. We observed a magnificent inhibition of tumor growth and metastasis in the combined treatment group over singular treatment groups in both models. This denotes that ARC2-001 largely augments the anti-cancer effect of widely employed anticancer drugs, such as 5FU, and thus can be exploited as an adjuvant for enhanced colorectal cancer chemotherapy.Collectively, our results evidenced the necessity of considering CAF-targeting agents for a more regulated therapeutic strategy to treat desmoplastic cancers. Citation Format: So-Young Yeo, Mohamed Farh, InSuk Sohn, Seok-Hyung Kim, Keun-Woo Lee. ARC2-001, a CAF-targeting drug exhibits strong anticancer effect in in vivo setting [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 LB322.

DNA-DEPENDENT PROTEIN KINASE INHIBITOR INDUCES APOPTOSIS IN COLON CANCER CELLS

Objectives: Among all types of DNA damage, DNA double-strand breaks (DSBs) are considered the most deleterious form induced by either endogenous factors (oxi-dative damages, mismatches, altered chromatin structures, and missing, or modified nucleotides) or exogenous factors, i.e., ultraviolet (UV) radiation, ionizing radiation (IR), and chemicals or drugs. DNA-dependent protein kinase (DNA-PK) plays a crucial role in repairing DSBs through non-homologous end joining (NHEJ). Cells lacking DNA-PK exhibit heightened sensitivity to IR and various DNA-damaging agents. The inhibition of DNA-PK further intensifies cellular susceptibility to IR and DNA-damaging agents. Several small molecules that inhibit DNA-PK have been developed. This study aimed to evaluate the effect of DNA-PK inhibitor (DNA-PKi) NU7441 on the HCT116 cell line. Methods: DNA-PKi NU7441 was used to assess the effect on anti-proliferation and induction of apoptosis on the HCT116 colorectal cancer cell line. Cells were cultured under standard conditions; crystal violet and apoptosis assay were applied to evaluate cell proliferation and apoptosis. Data were analysed using GraphPad Prism 8.4. Results: DNA-PKi effectively inhibited HCT116 colon cancer cell growth via crystal violet assay (p < 0.01). In addition, DNA-PKi also induced programmed cell death in the HCT116 cell line (p < 0.05). Conclusion: DNA-PKi NU7441 suppressed cell proliferation and induced apoptosis in the HCT116 colon cancer cell line.

In Vitro and In Silico Anticancer Evaluation of Secondary Metabolites from an Endophytic Fungus Aspergillus Fumigatus Isolated from Monarda Citriodora

AbstractChemical investigation of endophytic fungal extract of Aspergillus fumigatus strain KF436391 isolated from medicinal plant Monarda citriodora yielded one new compound 2‐methylenedecanedioic acid (1) along with seventeen known compounds (2–18) including (E)‐Undec‐2‐enedioic acid (2), Ergosterol (3) Ditryptophenaline (4), Gliotoxin (5), Pseurotin A (6), 12, 13‐dihydroxyfumitremorgin C (7), Brevianamide F (8), Demethoxy fumitremorgin C (9), Fumitremorgin C (10), Bisdethiobis (methylthio) gliotoxin (11), Cephalimysin A (12), Neofipiperazine C (13), Antibiotic FD838 (14), Monomethylsulochrin (15), trans‐p‐coumaric acid (16), trans‐ferulic acid (17) and 6‐Methoxyspirotryprostatin B (18). Anticancer evaluation study of all the compounds against a panel of eleven human tumor cell lines (PC‐3, HT‐29, MCF‐7, OVCAR‐5, HCT‐15, SF‐268, A549, HCT‐116, SW620, T47D and MD‐MBA‐435) of varied tissue origin revealed that compound 5 found to be the most potent cytotoxic agent against all the cancer cell lines examined, with an IC50 value <1.0 μM. But, it also showed cytotoxicity for normal human cells. Compounds 12, 14 and 15 exhibited maximum cytotoxicity against colon (HT‐29) cancer cell line with IC50 values 2.2, 2.8 and 7.7 μM respectively and were nontoxic for normal human cells. Docking studies of all the isolated compounds carried out using the Gaussian 09 suite of programs at B3LYP/6‐31+G(d) level of theory against three different proteins Poly [ADP‐ribose] polymerase‐1 (ID: 1UK0), TRAF2 (ID: 2X7F) and Matrix Metalloproteinase‐2 (ID: 1HOV) and maximum binding affinity was observed for TRAF2 (2X7F) protein. The in silico and in vitro anticancer results revealed that compounds 14 and 15 have the potential to be developed as anticancer agents against colon cancer cells.

Evaluation of In vitro Anti-Cancer Activity of Methanolic Leaf Extract of Phoenix pusilla on Colon Cancer Cell Line

ABSTRACT Background: Cancer rates continue to climb, owing largely to the world population’s aging and growth, as well as economically developing countries, a surge in cancer-causing behavior, particularly smoking. The third or fourth most prevalent type of cancer is colon cancer. Cancer of the large intestine (colon) is one of the primary causes of death from cancer. Colorectal cancer prevention is mostly based on adenomatous disease screening approaches. The cytotoxic and pharmacological properties of Phoenix pusilla are widely documented. As a result, there is little recorded evidence of its cytotoxic activity against colon cancer cells. Therefore, we planned to study the efficacy of a methanolic leaf extract of Phoenix pusilla against in vitro colon cancer cells. Aim: To evaluate the anti-cancer effects of the methanolic leaf extract of Phoenix pusilla on colon cancer cell lines. Materials and Methods: In vitro screening and anti-cancer effects of the methanolic effect of Phoenix pusilla on colon cancer cell lines were assessed by cell viability assays and cell and nuclear morphological studies. For the in vitro cell culture study, different concentrations of Phoenix pusilla leaf extract (0, 25, 50, 75, 100, 150 μg/ml) were used, and IC50 doses were calculated. Results: The results of the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay revealed that the fraction of viability cells significantly decreased in treated cells when compared to untreated control groups, was expressed as 100%, and an inhibitory concentration of μg/ml was identified. A phase-contrast microscope was used to observe cell shrinkage and cytoplasmic membrane blebbing. A fluorescent microscope was used to examine the apoptotic nuclei (internally dyed nuclei, shattered nuclei, and condensed chromatin). Conclusion: In conclusion, the present study results showed that the leaf extracts of Phoenix pusilla had a strong cytotoxic effect and induced significant apoptosis in the colon cancer cell lines at a concentration of 75 μg/ml in the 24 h incubation period. More research is needed to investigate the extract’s active components as well as the molecular mechanisms underlying its anti-cancer properties.

LncRNA ANRIL Promotes Glucose Metabolism and Proliferation of Colon Cancer in a High-Glucose Environment and is Associated with Worse Outcome in Diabetic Colon Cancer Patients.

The potential involvement of type 2 diabetes mellitus (T2DM) as a risk factor for colon cancer (CC) has been previously reported. Epigenetic changes, such as deregulation of long non-coding RNA (lncRNA) and microRNA (miR), have been linked to the advancement of CC; however, the effects of high glucose levels on their deregulation and, in turn, colon cancer remain unexplored. Fifty patients had a dual diagnosis of CC and T2DM, and 60 patients with CC without diabetes mellitus were included in the study. qRT-PCR was used to examine the expression of lncRNA ANRIL and miR-186-5p in tissue samples. ANRIL, miR-186-5p, and their downstream target genes HIF-1α, PFK, HK, Bcl-2, and Bax were also determined in CC cell lines under various glucose conditions. Glucose uptake, lactate production and cells proliferation were estimated in CC cell lines. A significant upregulation of ANRIL expression levels (p<0.001) and a significant downregulation of miR-186-5p expression (p<0.001) in diabetic colon cancer specimens compared to those in non-diabetic colon cancer group were observed. MiR-186-5p expression levels were inversely correlated with ANRIL expression levels, blood glucose levels and HbA1c%. Concerning in vitro model, a significant upregulation of ANRIL, downregulation of miR-186-5p, upregulation of HIF-1α, glycolytic enzymes and activation of antiapoptotic pathway was detected in higher glucose concentrations than lower one. There was a significant increase of glucose uptake, lactate accumulation and proliferation of the Caco2 and SW620 cell lines in a dose dependent manner of glucose concentrations. Moreover, a significant positive correlation between glucose uptake and ANRIL expression was shown. A high-glucose environment can increase the tumor-promoting effect of ANRIL. ANRIL can promote glucose metabolism and colon cancer proliferation by downregulating miR-186-5p with subsequent upregulation of glycolysis enzymes expression and inhibition of apoptosis.

Urolithin A affects cellular migration and modulates matrix metalloproteinase expression in colorectal cancer cells.

Colorectal cancer (CRC) is the world's second most common gastrointestinal malignancy. Preventing tumor cell proliferation and dissemination is critical for patient survival. Polyphenols have a variety of health advantages and can help prevent cancer. The current study examined different cellular activities of the gut-microbiota metabolite urolithin A (UA) on several colon cancer cell lines. The results revealed that UA suppressed cell growth in a dose- and time-dependent manner. In the current investigation, UA substantially affected cell migration in the wound-healing experiment and greatly decreased the number of colonies generated in each CRC cell culture. UA decreased cellular migration in CRC cells 48 h after treatment, which was significant (p < .001) compared to the migration rate in untreated cells. When compared to untreated cells, UA slowed the process of colony formation by reducing the number of colonies or altering their morphological shape. The western blot analysis investigation revealed that UA inhibits cellular metastasis by lowering the expression levels of matrix metalloproteinases 1 and 2 (MMP1 and MMP2) by more than 43% and 41% (p < .001) in HT29, 28% and 149% (p < .001) in SW480, and 90% and 74% (p < .001) in SW620, respectively, at a 100 µM dosage of UA compared to the control. Surprisingly, at a 100 µM dosage of UA, the expression levels of the tissue inhibitor of metalloproteinases 1 (TIMP1) were elevated in HT29, SW480, and SW620 cells treated with 100 µM of UA by more than 89%, 57%, and 29%, respectively. Our findings imply that UA has anticancer properties and might be used therapeutically to treat CRC. The findings provided the first indication of the influence of UA on cellular migration and metastasis in colon cancer cells. All of these data showed that UA might be used as an adjuvant therapy in the treatment of various forms of CRC.

Review of Sacred Herbal Medicine Aegle marmelos: A Potent Metastasis Inhibitor

Abstract: Aegle marmelos (A. marmelos) appears to be a significantly used ayurvedic medicine. This is a brilliantly composed nutritious fruit with carbohydrates, proteins, vitamins like riboflavin, thiamine, niacin, fatty acids, and minerals. Scientific studies have proved that A. marmelos has phytochemicals: carotenoids, phenolic, alkaloids, pectins, tannins, coumarins, flavonoids and terpenoids. Recent research on the effects of A. marmelos proved its anticancer, antimicrobial, cardioprotective, antidiabetic, and hepatoprotective activities. Extracts of various parts of plants such as leaves, bark, stems, fruits, and pulp subjected to preclinical studies conducted on cell lines showed apoptosis induction, inhibition of cell cycle and inhibition of proliferation of cells. Anti-cancer activity of A. marmelos was studied on selected cell lines of lung cancer, breast cancer, colon cancer and leukaemia. A. marmelos reported significant inhibitory effect on cell proliferation in cell line studies on A549, Ehrlich Ascites Carcinoma, MNU, DMBA, MCF-7, k562 cells, t-lymphoid Jurkat cells, b lymphoid Raji cells, erythroleukemic Hel cells, melanoma Colo 38 cells which supports the anticancer potential of it. Imperatorin has shown an antiproliferative effect on several cancer cell lines. The hydroethanolic extract of A. marmelos leaves showed a decrease in cell viability on A549 Human lung cancer cell line which works by fold change over control of the β-Catenin-m RNA and Wnt-mRNA expression of A549 and tumour growth suppression in DMBA induced carcinogenesis in rats. In human promyelocytic leukaemia, HL-60 cells, imperatorin from A. marmelos has caused cytochrome c-dependent apoptosis. Drugs interfere with the proliferative and survival signals that suppress the growth of K562 cells by blocking BCR-ABL1. Proper awareness, development of formulation and consumption of it make this drug scale up commercially.

Determination of Cytotoxic Compounds of Lepista personata (Fr.) Cooke by Gas Chromatography-Mass Spectrometry (GC-MS) and Chemometrics

The cytotoxic activities of hexane, acetone, and methanol extracts of Lepista personata were evaluated against lung (H1299), prostate (LNCaP), colon (CaCo-2), and breast (MCF-7) cancer cell lines. The hexane extract of Lepista personata exhibited significant cytotoxic activity against CaCo-2 (EC50: 198.7 ± 4.9 µg/mL), LNCaP (EC50: 152.1 ± 3.8 µg/mL), and MCF-7 (EC50: 98.37 ± 2.5 µg/mL). Only the hexane extract exhibited cytotoxicity; thus, it was fractionated to enrich the cytotoxicity against the studied cancerous cell lines over a silica gel column. Gas chromatography-mass spectrometry (GC-MS) analyses and the cytotoxicity of all fractions were investigated. Principal component analysis (PCA) was applied to the GC-MS and cytotoxicity (EC50) results. According to the PCA, the cytotoxic fractions containing abundant benzoic acid (0.01–26.19%), cinnamic acid (0.01–5.63%), arachidic acid (0.01–17.93%), heneicosanoic acid (0.01–14.57%), 1-monolinolein (0.01–26.34%), stigmasterol (0.01–10.15%) and ergosterol (0.01–17.10%) were clustered. Moreover, the chemical composition comparison of hexane extract and its fractions, when evaluated together with PCA, revealed that the bioactive compounds detected as benzoic acid, cinnamic acid, azelaic acid, 1-monolinolein, stigmasterol, and ergosterol, which were concentrated in fractions 3, 6, 7, and 9, were responsible for the cytotoxic activity. When used with bioactivity and chemometric analysis, GC-MS of the fractions identified the bioactive compounds.

Effects of down-regulated carbonic anhydrase 8 on cell survival and glucose metabolism in human colorectal cancer cell lines.

Carbonic anhydrase 8 (CA8) is a member of the α-carbonic anhydrase family but does not catalyze the reversible hydration of carbon dioxide. In the present study, we examined the effects of CA8 on two human colon cancer cell lines, SW480 and SW620, by suppressing CA8 expression through shRNA knockdown. Our results showed that knockdown of CA8 decreased cell growth and cell mobility in SW620 cells, but not in SW480 cells. In addition, downregulated CA8 resulted in a significant decrease of glucose uptake in both SW480 and SW620 cells. Interestingly, stable downregulation of CA8 decreased phosphofructokinase-1 expression but increased glucose transporter 3 (GLUT3) levels in SW620 cells. However, transient downregulation of CA8 fails to up-regulate GLUT3 expression, indicating that the increased GLUT3 observed in SW620-shCA8 cells is a compensatory effect. In addition, the interaction between CA8 and GLUT3 was evidenced by pull-down and IP assays. On the other hand, we showed that metformin, a first-line drug for type II diabetes patients, significantly inhibited cell migration of SW620 cells, depending on the expressions of CA8 and focal adhesion kinase. Taken together, our data demonstrate that when compared to primary colon cancer SW480 cells, metastatic colon cancer SW620 cells respond differently to downregulated CA8, indicating that CA8 in more aggressive cancer cells may play a more important role in controlling cell survival and metformin response. CA8 may affect glucose metabolism- and cell invasion-related molecules in colon cancer, suggesting that CA8 may be a potential target in future cancer therapy.

Engineering 5-flourouracil and leucovorin-loaded vesicular systems for possible colon specific delivery: In vitro evaluation and real time cell assay against HCT-116 colon cell lines

5-flourouracil (5-FU) is typically modulated with leucovorin (LEU) in clinical practice to improve clinical efficacy and patient survival rates. However, this combination has undesirable side effects and makes 5-FU more toxic. Hence, integrating a vesicular system (proniosomes) with another delivery vehicle may improve drug targeting, while resolving the aforementioned drawbacks. This study aimed to engineer 5-FU/LEU proniosomes for possible delivery to the colon. The modified slurry approach was used to create drug-loaded proniosomes (150 mg/9 g of carrier) using both water-soluble (dextrin (DEX) and lactose (LAC)) and insoluble (Neusilin FH2 (NEU)) carriers. The powdered formulations were filled into Eudragit S100 (10 %)-coated capsules or Eudragit FS 30D capsules for enteric- or colon-specific delivery. In vitro evaluations (flow properties, powder X-ray diffractometry (XRD) analysis, particle size analysis, entrapment efficiency, drug release, scanning electron microscopy (SEM), polydispersity index, Fourier transform infrared spectroscopy (FTIR), and stability studies) were performed on the formulations. An in vitro cytotoxicity test [real-time cell assay (RTCA)] against HCT-116 colon cancer cell lines was performed using the optimized formulation. In vitro evaluations showed that the nanoparticles had good physicochemical properties. RTCA studies showed sustained cell death with the formulations compared to the pure drug and placebo. The sequential drug release of the colon-targeted capsules containing 5-FU and LEU- loaded proniosomes showed negligible drug release in SGF (pH 1.2) and phosphate buffer solution (pH 6.8) (approximately 11 %) but profound drug release (>80 %) at pH 7.4. Drug-loaded proniosomes engineered for colon targeting (Eudragit S100 (10 %) capsules or Eudragit FS 30D capsules) showed good colon-specific targeting and favorable in vitro cytotoxicity profiles.

Exploring the antibiofilm effects on Escherichia coli biofilm associated with colon cancer and anticancer activities on HCT116 cell line of bee products

The association between dysbiotic microbiota biofilm and colon cancer has recently begun to attract attention. In the study, the apitherapeutic effects of bee products (honey, bee venom, royal jelly, pollen, perga and propolis) obtained from the endemic Yığılca ecotype of Apis mellifera anatoliaca were investigated. Antibiofilm activity were performed by microplate assay using crystal violet staining to measure adherent biofilm biomass of Escherichia coli capable of forming biofilms. Bee venom showed the highest inhibition effect (73.98%) at 50% concentration. Honey, perga and royal jelly reduced biofilm formation by >50% at all concentrations. The antiproliferation effect on the HCT116 colon cancer cell line was investigated with the water‑soluble tetrazolium salt‑1 assay. After 48 h of honey application at 50% concentration, cell proliferation decreased by 86.51%. The high cytotoxic effects of royal jelly and bee venom are also remarkable. Additionally, apoptotic pathway analysis was performed by ELISA using caspase 3, 8 and 9 enzyme-linked immunosorbent assay kits. All bee products induced a higher expression of caspase 9 compared with caspase 8. Natural products that upregulate caspase proteins are promising therapeutic targets for proliferative diseases.