HT-29 Human Colon Cancer Cells Research Articles

Chemical investigation and screening of anti-cancer potential of Syzygium aromaticum L. bud (clove) essential oil nanoemulsion.

This study was done to improve the medicinal properties of Syzygium Aromaticum L by processing S. Aromaticum L. bud essential oil (SABE) to the Nanoemulsion drug delivery system (SABE-NE) and investigating its anti-tumor and apoptotic impacts against the human HT-29 colon cancer cells. Applying the ultra-sonication method and characterization by DLS and FESEM analysis facilitates the nanoemulsification procedure. Human cancer (HT-29) and normal (HFF) cell lines were then evaluated based on the SABE-NE apoptotic and cytotoxic effects. In an in vitro section, flow cytometry method, Cas3 gene profile, AO/PI cell staining, and MTT assays are used to analyze the apoptotic and cytotoxic activities. In further analysis, liver lipid peroxidation and antioxidant genes expression (SOD, CAT, and GPx) investigate alterations in mice organs. As a result, produced 131.2nm SABE-NE induces apoptosis response and cellular death (Cas3 up-regulation and enhanced SubG1 peaks). Subsequently, the HT-29 cells' viability can reduce significantly, while HFF cells indicate confined cytotoxic impacts. Moreover, in vivo test results on mice livers demonstrate the cytoprotective properties of SABE-NE (reduced lipid peroxidation and increased antioxidant enzymes gene expression and nondetectable cytotoxic impacts). We produced a novel nanoemulsion drug delivery system called SABE-NE, a cell-specific apoptotic inducer. We thus can be utilized as an efficient anti-cancer compound for human colon cancer treatment. However, further supplementary studies are required to verify and approve its cell-specific anti-tumor activity.

Green synthesis and biological characterization of cerium oxide nanoemulsion against human HT-29 colon cancer cell line

ABSTRACT In this research, the cerium oxide nanoparticles (CeO2-NPs) were synthesized using Spirulina platensis microalgae. CeO2-NPs were nano-encapsulated based on ultrasonic emulsification method. Morphology, size, and stability of cerium oxide nanoemulsion (CeO2-NE) were verified by TEM, SEM, DLS, and zeta-potential. Antioxidant, antimicrobial against Escherichia coli and Staphylococcus aureus, and cytotoxic (on cancer cell line HT-29 and normal cell HFF) were performed comparatively between CeO2-NE and CeO2-NPs. Apoptotic and antioxidant related genes expression were determined in HT-29 treated cells with the CeO2-NE by Q-PCR. The CeO2-NE showed a much greater effect on anticancer activity, growth inhibition of two bacterial strains, antioxidant capacity, and reduction of ferric ions. Overexpression of caspase-3, caspase-8 and, caspase-9 of CeO2-NE confirmed apoptosis. Upregulation of superoxide dismutase attributed to the cellular redox potential of CeO2-NE in HT-29 cells. In conclusion, the CeO2-NE provides more efficient biological applications that could turn it into a promising factor for cancer treatment.

Mechanism of Pterostilbene-Induced Cell Death in HT-29 Colon Cancer Cells.

Pterostilbene is a dietary phytochemical that has been found to possess several biological activities, such as antioxidant and anti-inflammatory. Recent studies have shown that it exhibits the hallmark characteristics of an anticancer agent. The aim of the study was to investigate the anticancer activity of pterostilbene against HT-29 human colon cancer cells, focusing on its influence on cell growth, differentiation, and the ability of this stilbene to induce cell death. To clarify the mechanism of pterostilbene activity against colon cancer cells, changes in the expression of several genes and proteins that are directly related to cell proliferation, signal transduction pathways, apoptosis, and autophagy were also evaluated. Cell growth and proliferation of cells exposed to pterostilbene (5–100 µM) were determined by SRB and BRDU assays. Flow cytometric analyses were used for cell cycle progression. Further molecular investigations were performed using quantitative real-time RT-PCR. The expression of the signaling proteins studied was determined by the ELISA method. The results revealed that pterostilbene inhibited proliferation and induced the death of HT-29 colon cancer cells. Pterostilbene, depending on concentration, caused inhibition of proliferation, G1 cell arrest, and/or triggered apoptosis in HT-29 cells. These effects were mediated by the down-regulation of the STAT3 and AKT kinase pathways. It may be concluded that pterostilbene could be considered as a potential therapeutic option in the treatment of colon cancer in the future.

Effects of Sphingosine 1-Phosphate/Sphingosine-1-Phosphate Receptor on Pro-Angiogenesis in HT-29 Human Colon Cancer Cells

To investigate the effects and mechanism of sphingosine-1-phosphate/sphingosine-1-phosphate receptor on pro-angiogenesis in human colon cancer cells is the main objective. The effects of sphingosine-1-phosphate on the pro-angiogenesis in human colon cancer cells HT-29 and SW480 were assessed by tube formation assay. Quantitative real-time polymerase chain reaction was used to detect the expression of interleukin-8, interleukin-6 and vascular endothelial growth factor in HT29 and SW480 cells after sphingosine-1-phosphate treatment. Small interfering ribonucleic acid was designed to silence the expression of sphingosine-1- phosphate receptor 1, sphingosine-1-phosphate receptor 2 and sphingosine-1-phosphate receptor 3 in HT-29 and SW480 cells. Western blot and quantitative real-time polymerase chain reaction were used to assess the down-regulation efficiency of sphingosine-1-phosphate receptor in HT-29 and SW480 cells. Later, the effects of sphingosine-1-phosphate receptor were detected by quantitative real-time polymerase chain reaction. Tube formation assay showed that the number of formed tubes in human umbilical vein endothelial cell-fused cells (EA.hy926) resuspended in the culture supernatant from HT-29 and SW480 cells treated with sphingosine-1-phosphate was significantly more than that of the control group (t=3.667 and 4.881, p=0.021 and 0.013), indicating that sphingosine-1-phosphate promoted the pro-angiogenic ability of colon cancer cells. Moreover, the messenger ribonucleic acid expression of interleukin-8, interleukin-6 and vascular endothelial growth factor were significantly increased after sphingosine-1-phosphate treatment compared to the control group (p<0.05) where sphingosine-1-phosphate receptor 1 gene silence could significantly decrease the messenger ribonucleic acid expression in HT-29 and SW480 cells (p<0.05), while sphingosine-1-phosphate receptor 2 and sphingosine-1-phosphate receptor 3 gene silence did not lead to significant messenger ribonucleic acid changes. Sphingosine-1-phosphate up-regulated the expression of interleukin-8, interleukin-6 and vascular endothelial growth factor in human colon cancer cells through sphingosine-1-phosphate receptor 1 and thereby enhancing the proangiogenic effect of colon cancer cells. The sphingosine-1-phosphate/sphingosine-1-phosphate receptor pathway is a promising novel therapeutic target for inhibiting colon cancer growth.

Genomic and Phenotypic Characteristics in Geographically Separated Clinical Campylobacter jejuni ST353CC Isolates.

Campylobacter jejuni fecal isolates of eight international travelers, 5 of which had traveled to Ecuador and 3 to Bangladesh, were characterized, and the possible relationship between bacterial traits and clinical symptoms was further analyzed. All eight isolates belonged to the same Multi-Locus Sequence Type clonal complex (ST353CC). The three isolates from Bangladesh were all of the same sequence type (ST-9438), and when compared to isolates of various other sequence types, they had a larger quantity of unique genetic content, higher expression levels of some putative virulence genes involved in adhesion and invasion (flpA, ciaB and iamA), and showed higher adhesion levels to human HT-29 colon cancer cells in an in vitro infection model. However, in contrast to the seemingly higher pathogenic potential of these bacterial isolates, travelers infected with the ST-9438 isolates had no or only very mild symptoms, whereas the other individuals, whose bacterial isolates seemed to have less pathogenic potential, generally reported severe symptoms. When studying the 16S rRNA gene-based fecal microbiota in samples collected prior to travel, there was an individual variation in the relative abundance of the three major bacterial phyla Actinobacteria, Bacteroidetes and Firmicutes, but there were no associations between composition and diversity of microbiota and development of severe symptoms from the infection. It remains to be confirmed by larger studies whether an individual’s characteristics such as gut microbiota, might be related to the severity of symptoms in Campylobacter infections.

Antioxidant and Anti-Inflammatory Activity of Coffee Brew Evaluated after Simulated Gastrointestinal Digestion.

Coffee contains human health-related molecules, namely polyphenols that possess a wide range of pharmacological functions, and their intake is associated with reduced colon cancer risk. This study aimed to assess the changes in the anti-inflammatory and antioxidant activity of coffee after simulated gastrointestinal digestion. The evaluation of intracellular reactive oxygen species (ROS) levels in the HT-29 human colon cancer cell line and three in vitro spectrophotometric assays were performed to determine the antioxidant activity of the samples. Characterization of coffee composition was also assessed through a Q-Orbitrap high-resolution mass spectrometry analysis. The results highlighted that the levels of polyphenols in the digested coffee brews were higher than those of the non-digested ones. All assayed samples decreased the levels of intracellular ROS when compared to untreated cells, while digested coffee samples exhibited higher antioxidant capacity and total phenolic content than not-digested coffee samples. Digested coffee samples showed a higher reduction in interleukin-6 levels than the not-digested samples in lipopolysaccharide-stimulated HT-29 cells treated for 48 h and fewer cytotoxic effects in the MTT assay. Overall, our findings suggest that coffee may exert antioxidant and anti-inflammatory properties, and the digestion process may be able to release compounds with higher bioactivity.

Real-Time Impedance Monitoring of Alkaline Phosphatase (ALP) Release by Microelectrode Arrays

Alkaline phosphatase (ALP) is a reporter enzyme widely used in point-of-care applications. Real-time monitoring of ALP accelerates understanding of emerging diseases and of cancer progression. This can be achieved through biosensor-based strategies. This paper outlines an approach that facilitates the real-time monitoring of ALP release from the differentiation of cultured human colon cancer cells (Ht-29). Electric cell-substrate impedance sensing was used to generate changes in Ht-29 cells during ALP release and the data were compared using amperometry. The findings indicate a correlation between cell response and shift in impedance magnitude and current. Relevant results had similar attitude during cell monitoring, which influenced cell morphology and cell viability. Our analysis is a clear advance on the current methods such as enzyme-linked immunosorbent assay. The developed method can contribute to the development of microfluidic testing that would facilitate work on small-scale techniques.

Regulation of Butyrate-Induced Resistance through AMPK Signaling Pathway in Human Colon Cancer Cells.

Butyrates inhibit cell growth in colon cancer cells by inhibiting histone deacetylases. However, chronic exposure to butyrates induces butyrate resistance in colon cancer cells. The mechanism underlying the acquisition of resistance is not yet fully understood. Here, butyrate-resistant (BR) colon cancer cells were developed in HCT116, HT29, and SW480 human colon cancer cells and were confirmed by the increase in the inhibitory concentrations of cell growth by 50% (IC50) compared to their respective parental (PT) cells. Chronic exposure to butyrate induced autophagy via higher expression of Beclin-1 and LC3B-II. The AMP-activated protein kinase (AMPK) was downregulated along with the activation of Akt and mammalian target of rapamycin (mTOR) and decrease in acetyl-CoA carboxylase (ACC) in BR colon cancer cells compared to those in their respective PT cells. Activation of AMPK by AICAR treatment in BR colon cancer cells suppressed cell proliferation by inhibiting Akt and mTOR and activating ACC. Taken together, chronic exposure to butyrate increased butyrate resistance in human colon cancer by inducing protective autophagy through the downregulation of AMPK/ACC and activation of Akt/mTOR signaling. Activation of AMPK restored sensitivity to butyrate by the inhibition of Akt/mTOR, suggesting that AMPK could be a therapeutic target for BR colon cancers.

The new andrographolide derivative AGS-30 induces apoptosis in human colon cancer cells by activating a ROS-dependent JNK signalling pathway

Background: The anti-cancer activity of andrographolide (Andro) has been extensively demonstrated in recent years. It is supposed that modifying the chemical structure of Andro can improve its efficacy and reduce its toxicity. Purpose: In this study, the anti-cancer effect of a 14β-(2′-chlorophenoxy) derivative of andrographolide known as AGS-30 was investigated, and its underlying mechanisms were also explored. Study Design/Methods: Different cancer cells were used to evaluate and compare the in vitro anti-cancer effects of Andro and AGS-30. Human colon cancer cells HT-29 and HCT-116 were used to study the underlying anti-cancer mechanisms of AGS-30. HT-29 cells xenografted in nude mouse model was used to compare the in vivo anti-tumour efficacies of Andro and AGS-30. Result:In vitro studies showed that AGS-30 possessed an anti-cancer effect by inhibiting the viability, colony formation and migration of cancer cells. It significantly induced the generation of reactive oxygen species (ROS), caused the loss of mitochondrial membrane potential and triggered the apoptosis in colon cancer cells. These effects of AGS-30 were more potent than those of Andro. In addition, the expression levels of proteins associated with apoptosis, including phospho-JNK1/2 as well as cleaved caspase 9, caspase 3, and poly(ADP ribose) polymerase, were elevated in AGS-30-treated colon cancer cells. Moreover, these elevated levels of the proteins were inhibited by the antioxidant N-acetylcysteine and the JNK inhibitor SP600125, suggesting the involvement of ROS/JNK-dependent mechanisms in AGS-30-induced apoptosis. The in vitro anti-cancer effect could be reproduced in an HT-29 colon cancer cell xenografted nude mouse model. Conclusion: The anti-cancer effect of AGS-30 is stronger than that of Andro. AGS-30 induces apoptosis of colon cancer cells through ROS/JNK-dependent pathway. Our findings may provide insights for the future development of derivatives of Andro as novel chemotherapeutic agents.

A bis indole analog molecule fluorescent probe for living cell staining

In this study, the fluorescent probe property of a bis indole compound (5,14-dimethyl-5,6,7,8,9,14-hexahydroindolo [2 ', 3′: 3,4] cycloocta [1,2-b] indole), a Caulerpin derivative, which is a member of the indole family, was investigated. For this purpose, firstly, the fluorescence character of the molecule was investigated by absorption and fluorescence spectroscopy techniques in both solvent and surfactant environments. Afterwards, the cytotoxicity of the molecule was tested on the human colon cancer HT-29 cells and breast cancer MCF-7 cells as well as mouse fibroblast 3T3-L1 cells. After determining that the molecule has no cytotoxic effect against all the tested cell lines by 100 µM, the cell staining feature of the probe was examined. The obtained data showed us that the molecule, whose confocal microscopy images were taken at 360 nm excitation wavelength, stained the cell cytoplasm. In addition, optical absorption spectra, oscillator strength (f), transition configuration and transition characteristics of the Ars-In in different solvents were investigated at the theoretical level using Time Dependent-Density Functional Theory (TD-DFT). In conclusion, these results confirm that this indole derivative molecule could be evaluated as an effective fluorescence probe for biological systems.

Biomedical potential of silver nanoparticles capped with active ingredients of Hypnea valentiae, red algae species

In this present study, marine red algae Hypnea valentiae capped silver nanoparticles (Hv-AgNPs) have been prepared through the green method and characterized by various analytical tools. The prepared Hv-AgNPs were confirmed by Ultraviolet-visible (UV) spectroscopy at 430 nm. The X-ray diffraction (XRD) analysis affirmed the face-centered cubic crystalline structure of Hv-AgNPs. Transmission electron microscope (TEM) revealed the Hv-AgNPs shape as spherical with an average size of 10–45 nm. Fourier Transform Infrared (FR-IR) analysis depicted the presence of various biomolecules such as carboxyl, phenolic, and amine groups, which acted as reducing and capping agents of biosynthesized silver nanoparticles. The Hv-AgNPs had significant antibacterial activity against four bacterial pathogens. The 2,2’-Diphenyl-1-picrylhydrazy (DPPH) assay revealed the strong free radical scavenging ability of Hv-AgNPs. The Hv-AgNPs exhibited excellent anti-cancer activity against HT-29 human colon cancer (IC50–24.6 μg/ml) and A549 lung cancer cell lines (IC50–5.917 µg/ml) by MTT assay. Furthermore, the investigations demonstrated that green synthesized silver nanoparticles possess high anti-bacterial, anti-oxidant and anti-cancer activities.

Evaluation of Radiosensitive Effect of Tolmetin in Radiotherapy on Human Colonic Carcinoma Cell Line HT-29

Background: Radiotherapy is one of the cancer treatment methods. Although radioresistance and normal tissue toxicity limit the radiation therapy in certain anatomical locations, using some substances can be useful to increase radiosensitivity on cancer cells without cytotoxicity effect on normal cells. Objective: This study aimed to evaluate the radiosensitizing effect of tolmetin in radiotherapy treatment on human colon cancer cell line HT-29. Methods In this study, human colon cancer cells HT-29 in different groups were irradiated with 4 Gy x-ray, and tolmetin was administered in different concentrations (75, 100, and 150 μM). Then, the groups were compared with each other and with the control group by Micronucleus Assay (MN) and Nuclear Division Index (NDI). NDI investigated the cytotoxicity, and MN indicated the genotoxicity. Results: In the group receiving radiation, micronuclei increased significantly compared to the control group. In the group receiving tolmetin with a concentration of 75 and 100 μM, the number of micronuclei also increased compared to the control group. In all groups treated with tolmetin that also received radiation, a significant increase in micronuclei was observed, which was more noticeable at concentrations of 100 and 150 μM. At the same time, tolmetin at the studied concentrations did not change the NDI index. Conclusion: The present study demonstrates that tolmetin has a radiosensitizing effect on HT-29 colon cancer cells, which depends on the tolmetin concentration. In addition, tolmetin has no cytotoxic effect on this cell line.

Synergistic Anti-proliferative Effects of Lenalidomide and Dexamethasone on the HT-29 Cell Line Through Apoptotic Genes

Background: Colorectal cancer (CRC) is the third most common cancer among men and the second most common type of cancer among women worldwide. The resistance of tumor cells to apoptosis is caused by changes in the expression of anti-apoptotic or pro-apoptotic proteins. Histone deacetylase inhibitors (HDACi) are known to cause changes in gene expression. Objectives: The present study aimed at investigating the anti-proliferative effects of lenalidomide (LEN) as HDACi and dexamethasone (DEX) on the human colon cancer HT-29 cell line. Methods: The HT-29 cell line was treated with various concentrations of LEN and DEX individually and in combination for 24, 48, and 72 hours. Cytotoxicity was evaluated by MTT assay. The half-maximal inhibitory concentration (IC50) was measured, and quantitative real-time polymerase chain reaction (qRT-PCR) was also performed to examine the expression of Bcl2, Bax, Fas, and FasL genes. Results: The combination of LEN (1000 µM) with DEX (100 µM) showed potent synergistic anti-proliferative activities in a time- and dose-dependent manner. The combination of these drugs induced cell death by affecting the extrinsic and intrinsic apoptotic gene expression profiles. Conclusions: The combination of LEN with DEX can be proposed as a new therapeutic approach for CRC.

Biocompatible Eu doped mesoporous calcium silicate nanospheres for pH-responsive drug release

Calcium silicate is a kind of inorganic biomaterial containing Si, Ca, O elements, they have been widely investigated and applied in a variety of medical applications in the past four decades. Herein, Europium (Eu) doped mesoporous calcium silicate nanoparticles (Eu-MCS) with sizes ranging from 60 nm to 110 nm were prepared via a simple hydrothermal synthesis method. The morphology, structure, texture and optical property were characterized by SEM, TEM, XRD, N2 adsorption/desorption and PL spectra, respectively. The prepared material emits strong red luminescence at 613 nm upon UV irradiation and is cytocompatible at a wide range of concentrations, exibiting promising application in bioimaging. The Eu-MCS has a high loading capacity of 356.67 μg/mg for the anticancer drug DOX, and the drug loaded material DOX@Eu-MCS showed pH-controlled drug release behaviour. Moreover, both of the biocompatibility tests on normal human cells HEK293 and human colon cancer cells HT29 indicate a low cytotoxicity of Eu-MCS. The cell cytotoxity assays of the DOX loaded nanomaterial (DOX@Eu-MCS) on Hela cancer cells prove a high cancer cell mortality. Therefore, the as-prepared red emissive nanocarrier Eu-MCS has great potential to be used as an integrated platform for cancer diagnosis and therapy.

In vitro antimetastatic activity of Momordica balsamina crude acetone extract in HT-29 human colon cancer cells.

Plant-derived compounds and/or extracts have proven to be beneficial for the treatment of a broad spectrum of cancers with minimal side effects. In this study, we investigated whether a crude acetone extract of Momordica balsamina (MBE) can interfere with the metastatic ability of HT-29 colorectal cancer (CRC) cells. The phytochemical composition of MBE was determined by ultra-performance liquid chromatography and cytotoxic effects by the MTT and acridine orange/ethidium bromide staining assays. The effect of MBE on the formation of reactive oxygen species was assessed using the DCFH2 -DA assay. Wound healing assay, transwell cell invasion assay, cell adhesion assay, and the extracellular matrix-cell adhesion array were used to assess the antimetastatic effects of MBE. The effect of MBE on the expression of TNF-α, NF-κB, TIMP-3, MMP-2, and MMP-9 was assessed by western blot analysis. Our results showed that MBE consists of a mixture of compounds without a known anticancer activity in CRC and exhibits cytotoxicity against HT-29 cells. MBE also suppressed reactive oxygen species formation, cell invasion, cell migration, and cell adhesion. The reduction of cell invasion was associated with the downregulation of TNF-α, NF-κB, MMP2, and MMP9 and upregulation of TIMP-3 proteins. We concluded that MBE inhibits the metastatic ability of HT-29 CRC cells in vitro.

Metabolite Profiling of Rosemary Cell Lines with Antiproliferative Potential against Human HT-29 Colon Cancer Cells.

Rosemary (Rosmarinus officinalis) is a culinary and medicinal plant used in food and pharmaceutical industry. The wide range of biological activities is mainly related to phenolic and terpenic compounds; like carnosic acid (CA), carnosol (CS) and rosmarinic acid (RA), mainly reported in rosemary leaf extracts, and recently described in rosemary callus extracts. The aim of this work was to investigate the chemical profile of rosemary cell lines and evaluate their antiproliferative potential against human HT-29 colorectal cancer cell lines. For this purpose, rosemary leaf explants were dedifferentiated on MS medium and added with 2, 4-D (2, 4-dichlorophenoxyacetic acid; 2mg/L) and BAP (6-benzylaminopurine; 2mg/L). Cell aggregates were separated according to colour and three rosemary cell lines cultures were established: green (RoG), yellow (RoY) and white (RoW).The chemicalprofile of rosemary cell lines extracts was characterized by combining HPLC and GC platforms coupled to HR-MS/MS. The antiproliferative activityagainst HT-29 cell line was analyzed with MTT assay. A total of 71 compounds, including hydroxycinnamic acid and hydroxybenzoic acid derivatives,flavonoids, phenolic di- and triterpenes, as well as relevant unsaturated fatty acids and their esters, phytosterols, and carotenoids were tentatively identifiedin the extract of the target cell lines. The antiproliferative activity test against HT-29 cell using the MTT assay revealed that the viability of HT-29 coloncancer cells was affected after treatment with the RoW extract (IC50 of 49.63 μg/mL) at 48 h. These results showed that rosemary cell lines can alsoaccumulate other bioactive phytochemicals with demonstrated antiproliferative potential.

Evaluation of the Effect of Valproic Acid on JAK/STAT Pathway, SOCS1, SOCS3, Bcl-xL, c-Myc, and Mcl-1 Gene Expression, Cell Growth Inhibition and Apoptosis Induction in Human Colon Cancer HT29 Cell Line

Evaluation of the Effect of Valproic Acid on JAK/STAT Pathway, SOCS1, SOCS3, Bcl-xL, c-Myc, and Mcl-1 Gene Expression, Cell Growth Inhibition and Apoptosis Induction in Human Colon Cancer HT29 Cell Line

Abstract 1069: Determining the effect of bazedoxifene in combination with chemotherapy on colon cancer cells

Abstract Inflammatory cytokines, such as interleukin-6 (IL-6) and IL-11, activate the signal transducer and activator of transcription 3 (STAT3), which drives the gene transcription of cellular processes attributed as hallmarks of cancer. High levels of these cytokines, in combination with driver mutations, facilitate tumour growth and progression in preclinical models of colon cancer. Previous studies in our laboratory identified bazedoxifene (BZA), currently approved for the treatment of osteoporosis, as a small molecule inhibitor of GP130 (the receptor common to the IL-6 family of cytokines), selectively suppressing IL-6 and IL-11 signalling to reduce colon and gastric cancer growth in vivo (Thilakasiri et al. EMBO Mol Med 2019). The aim of this study is to determine the key cellular mechanisms influenced by GP130 inhibition to mediate anti-tumour effects in colon cancer cells. Additionally, we assessed the combined effects of BZA with standard of care chemotherapy drugs on cell proliferation and apoptosis. Using LIM2405 and HT29 colon cancer cells, the combined effect of BZA, fluorouracil and oxaliplatin on inducing apoptosis in LIM2405 and HT29 colon cancer cells was analysed using flow cytometry and confirmed using Western blotting and DIC microscopy. The effects of BZA was further analysed on HT29 xenograft growth and metastasis in vivo. Levels of STAT3 activation in response to IL-11/STAT3 signalling was characterised by Western blotting. Our results demonstrated that combination treatment with BZA, fluorouracil and oxaliplatin significantly increased apoptosis in LIM2405 cells and BZA significantly induced apoptosis in HT29 cells. Our data showed that BZA inhibited GP130-dependent STAT3 activity in the human colon cancer cell line LIM2405 and HT29. BZA treatment sensitized cells to chemotherapy leading to increased apoptosis. BZA treatment also reduced both primary tumour and metastatic burden in vivo demonstrating that gp130 mediated STAT3 inhibition is an attractive target for the treatment of early stage and advanced stage colon cancer. Citation Format: Rhynelle S. Dmello, Pathum Thilakasiri, Belinda Duscio, Ivan Poon, Matthias Ernst, Ashwini Chand. Determining the effect of bazedoxifene in combination with chemotherapy on colon cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1069.

Xanthatin inhibits human colon cancer cells progression via mTOR signaling mediated energy metabolism alteration.

Tumor cells exhibit higher glycolysis and rely on abnormal energy metabolism to produce ATP, which is essential for cell proliferation and migration. Abnormal energy metabolism inhibition is considered a promising tumor treatment strategy. Xanthatin is an active sesquiterpene lactone isolated from Xanthium strumarium L. This study evaluated the effect of xanthatin on the energy metabolism of human colon cancer cells. The results showed that xanthatin significantly inhibited the migration and invasion of human HT-29 and HCT-116 colon cancer cells. We found that xanthatin effectively reduced the production of ATP and promoted the accumulation of lactate. Xanthatin inhibited glycolysis which may be related to the reduction of glucose transporter 1 (Glut1) and monocarboxylate transporter 4 (MCT4) mRNA and protein levels. Concomitantly, xanthatin promoted complex II activity and oxidative phosphorylation (OXPHOS), resulting in mitochondrial damage and cell death in HT-29 cells. Furthermore, xanthatin inhibited the phosphorylation of mTOR, the phosphorylation of 4E-binding protein 1 (4E-BP1) and c-myc in HT-29 cells. Moreover, rapamycin, a mTOR inhibitor, could enhance the cytotoxicity effect in xanthatin treated HT-29 cells. Additionally, HT-29 cells transfected with si-mTOR aggravated xanthatin induced cell viability inhibition. Based on these results, we observed that the effect of xanthatin on energy metabolism may be related to its inhibition of the mTOR signaling pathway. Collectively, this study provides important insights into xanthatin's anticancer effect, which occurs by regulation of the energy metabolism of human colon cancer cells, and suggest that xanthatin has potential as a botanical drug against abnormal tumor energy metabolism.

Functional Properties of Chlorella vulgaris, Colostrum, and Bifidobacteria, and Their Potential for Application in Functional Foods

The market for new functional foods and food supplements is rapidly evolving, with a current emphasis on using natural sources. Algae, probiotics, and colostrum are rich sources of nutrients and bioactive compounds with positive effects on human and animal health. To determine the potential for developing new functional foods combining these components, we evaluated their synergistic effects. We assessed the growth of selected bifidobacteria in a medium supplemented with Chlorella vulgaris and its immunomodulatory and cytotoxic effects on the human peripheral mononuclear cells and colon cancer cell lines Caco-2 and HT29. The hypocholesterolemic effects of Chlorella powder and bovine colostrum fermented by Bifidobacterium animalis subsp. lactis BB12® on lipid metabolism in rats fed a high-fat diet were also determined. Chlorella addition promoted Bifidobacteria growth, with significantly increased inflammatory cytokine (TNF-α and IL-6) levels following 1.0% (w/v) Chlorella stimulation. Rats fed diets containing fermented colostrum with 0.5% (w/v) added Chlorella powder exhibited significantly decreased triglyceride, very low-density lipoprotein, and alanine and aspartate aminotransferase levels, compared to those of the control group. These results support that C. vulgaris is not cytotoxic in intestinal cell models and affords prebiotic and immunomodulatory effects, as well as synergistic triglyceride-lowering effects with bovine colostrum and B. animalis subsp. lactis BB-12.