Colon Cancer Cell Death Research Articles

The Suppressive Effect of Novel Hydrazones‐Tethered Imidazoles in HCT‐116 and HT‐29 Colorectal Cancer Cells: Synthesis, Biological Activity and Molecular Modeling Studies

AbstractA series of hybrid compounds containing both the imidazole ring and the hydrazone moiety have been synthesized. Synthesized compounds were characterized by various spectral techniques, including FT‐IR, 1H‐NMR, 13C‐NMR, and HRMS. The compounds were evaluated for their antiproliferative activities on colorectal cancer cells HCT‐116 and HT‐29 in a time‐dependent manner. Among them, some compounds exhibited remarkable anti‐cancer activity with a less cytotoxic effect on non‐cancerous cell lines, especially HRK‐2 with IC50 value of 1.35±0.18 μM in HCT‐116 cells and HRK‐5 with IC50 value of 2.67±0.61 μM in HT‐29 cells. Investigations of colon cancer cell death were performed, and the most active compounds were found to trigger cell death via nuclear localization and induce S phase arrest of the colon cancer cell. Moreover, molecular modeling studies for the synthesized compounds was performed to predict their binding affinities toward the active site of BCL‐2. The findings of the molecular modeling investigations were highly consistent with those of the cytotoxicity results.

Phytocannabinoids CBD, CBG, and their Derivatives CBD-HQ and CBG-A Induced In Vitro Cytotoxicity in 2D and 3D Colon Cancer Cell Models.

Phytocannabinoids, compounds found in Cannabis sativa L., are used in oncology and palliative care to reduce the adverse reactions of standard therapies. Cancer patients use formulations of Cannabis sativa L. to manage the anxiety, pain, and nausea associated with cancer treatment, and there is growing evidence that some of them may exhibit anticancer properties. In this study, we tested the anticancer potential of selected cannabinoids CBD (cannabidiol) and its quinone derivative CBD-HQ (cannabidiol hydroquinone), CBG (cannabigerol) and its acid derivative CBG-A (cannabigerolic acid), as well as a combination of CBD+CBG on the colon cancer cell line SW-620. The MTT assay was used to determine the cannabinoids' ability to induce colon cancer cell death. All cannabinoids were cytotoxic at the lowest concentration (3 μg/mL). The half maximal inhibitory concentration (IC50) ranged from 3.90 to 8.24 μg/mL, depending on the substance. Cytotoxicity was confirmed in a 3D spheroidal cell culture with calcein and propidium iodide staining. The amount of intracellular reactive oxygen species (ROS) was examined using a DCF-DA assay. CBG showed the lowest antioxidant activity of all the cannabinoids tested. The level of intracellular ROS decreased only by 0.7-18%. However, CBG-A induced the strongest reduction in ROS level by 31-39%. Our results suggest that cannabinoids represent an interesting research direction with great implementation potential. These preliminary results represent the beginning of research into the potential of these substances for anticancer treatment and underscore the potential for further research.

Ginger Root Bioactive Compounds Specifically Inhibits Growth of Colon Cancer Cells in Culture.

Colon cancer is affluent among many people, and having cancer greatly impacts the lives of many. Ginger is a common food, particularly in Asian cuisine. However, the health benefits of ginger as a whole food and 6-gingerol, its bioactive compound in prevention of colon cancer have not been fully addressed. This experiment investigated effects of ginger juice and 6-gingerol on colon cancer cell growth and death. Fresh ginger roots were homogenized for juice preparation. Total phenolic contents of ginger juice were measured using Folin-C assay. Colon cancer SW480 cells and normal colon epithelial cells CCD-18Co were treated with ginger juice and/or 6-gingerol. Cell metabolic activity was assessed by MTT assay. Cell apoptosis and cell cycle arrest were accessed by immunoblotting. Data were analyzed by 2-way ANOVA with a Tukey post-hoc test and statistical significance was set at P < .05. The results showed that ginger juice selectively inhibited SW480 cell growth at 25 µL/mL for 40 hours. High doses of ginger juice (at 50 and 100 µL/mL for 40 hours) inhibited the growth of both cell types. This was independent of caspase-3 activation. Six-gingerol specifically inhibited SW480 cell growth starting at 0.5 µmoL/L (P < .01). More than 1 µmoL/L 6-gingerol did not give more power to inhibit SW480 cell growth. The results also showed that CCD-18Co cell growth rates were not changed after 6-gingerol treatments (up to 10 µmoL/L, P > .1). Immunoblotting results revealed that the elevation of Myt1 levels and decreases in CDK1, p21 Wafl/Cip1 and pSer642-Wee1 only occurred in SW480 but not CCD-18Co cells when treated with 1 and/or 2.5 µmoL/L 6-gingerol for 40 hours. 6-gingerol can specifically inhibit SW480 cancer cells without killing normal CCd-18Co cells, through cell cycle arrest. Ginger juice can selectively inhibit colon cancer cell growth in a narrow window at ~25 µL/mL.

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

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

Adenosine dialdehyde, a methyltransferase inhibitor, induces colorectal cancer cells apoptosis by regulating PIMT:p53 interaction

Post-translational modification (PTM) is important in controlling many biological processes by changing the structure and function of a protein. Protein methylation is an important PTM, and the role of methyltransferases has been implicated in numerous cellular functions. Protein L-isoaspartyl methyltransferase (PIMT) is ubiquitously expressed in almost all organisms and govern important cellular processes including apoptosis. Among other functions, PIMT has also been identified as a potent oncogene because it destabilizes the structure of the tumor suppressor p53 via methylation at the transactivation domain. In the present study we identified that out of the three methyltransferase inhibitors tested, namely, S-adenosyl-l-homocysteine (AdoHcy), adenosine and adenosine dialdehyde (AdOx), only AdOx augments p53 expression by destabilizing PIMT structure, as evident from far-UV CD. The effect of the inhibitors, AdOx in particular, to the structure of PIMT, and the binding of PIMT to the p53 transactivation domain have been investigated by docking and molecular dynamics simulations. AdOx significantly increases p53 accumulation and nuclear translocation in colon cancer cells, triggering the p53-mediated apoptotic pathway. To better understand the molecular mechanisms underlying p53 accumulation in colon cancer cells, we observed that the level of PIMT is considerably lower in AdOx-treated cells, reducing its association with p53, which stabilized p53. p53 then transactivated BAX, increasing the BAX: BCL-2 ratio and causing colon cancer cell death.

Reactive oxygen species mediated apoptotic death of colon cancer cells: therapeutic potential of plant derived alkaloids.

Colorectal cancer (CRC) is one of the most deaths causing diseases worldwide. Several risk factors including hormones like insulin and insulin like growth factors (e.g., IGF-1) have been considered responsible for growth and progression of colon cancer. Though there is a huge advancement in the available screening as well as treatment techniques for CRC. There is no significant decrease in the mortality of cancer patients. Moreover, the current treatment approaches for CRC are associated with serious challenges like drug resistance and cancer re-growth. Given the severity of the disease, there is an urgent need for novel therapeutic agents with ideal characteristics. Several pieces of evidence suggested that natural products, specifically medicinal plants, and derived phytochemicals may serve as potential sources for novel drug discovery for various diseases including cancer. On the other hand, cancer cells like colon cancer require a high basal level of reactive oxygen species (ROS) to maintain its own cellular functions. However, excess production of intracellular ROS leads to cancer cell death via disturbing cellular redox homeostasis. Therefore, medicinal plants and derived phytocompounds that can enhance the intracellular ROS and induce apoptotic cell death in cancer cells via modulating various molecular targets including IGF-1 could be potential therapeutic agents. Alkaloids form a major class of such phytoconstituents that can play a key role in cancer prevention. Moreover, several preclinical and clinical studies have also evidenced that these compounds show potent anti-colon cancer effects and exhibit negligible toxicity towards the normal cells. Hence, the present evidence-based study aimed to provide an update on various alkaloids that have been reported to induce ROS-mediated apoptosis in colon cancer cells via targeting various cellular components including hormones and growth factors, which play a role in metastasis, angiogenesis, proliferation, and invasion. This study also provides an individual account on each such alkaloid that underwent clinical trials either alone or in combination with other clinical drugs. In addition, various classes of phytochemicals that induce ROS-mediated cell death in different kinds of cancers including colon cancer are discussed.

An ellagitannin-loaded CS-PEG decorated PLGA nano-prototype promotes cell cycle arrest in colorectal cancer cells

Colorectal cancer is associated with significant morbidity and mortality worldwide. Egypt, as a developing country, has a high-rise incidence of cancer. The current study objective was to investigate the antitumor influences of ellagitannin-loaded CS-PEG-decorated PLGA nano-prototypes against human colorectal cancer cell lines (HCT 116 as well as Caco-2) in vitro. Doxorubicin (DOX), punicalin (PN), and punicalagin (PNG)-encapsulated chitosan-polyethylene glycol-decorated PLGA (PLGA-CS-PEG) nanoparticles (NPs) were described. The cytotoxicity of each preparation was evaluated using MTT assays in HCT 116 as well as Caco-2 cells during G0, G1, S, and G2 cell cycle phases. Cell cycle-related gene expression and protein levels were measured after treatment. Reactive oxygen species (ROS) levels were also measured. Both PN and PNG PLGA-CS-PEG NPs induce colon cancer cell death with cell cycle arrest in the G1 phase in vitro. Caco-2 cells were more sensitive to the nano-therapy than HCT 116 cells. Upon treatment, the ratio of Bax to Bcl-2 expression was increased following nano-therapy, with increased levels of Cas-3 and decreased expression of Bcl-2, PI3k, and NF-ĸB compared to control. The nitric oxide level (NO), a marker of ROS, was increased following nano-therapy compared to control. In conclusion, ROS-mediated cell cycle arrest can be induced by PN as well as PNG nano-therapy in cell lines of colorectal cancer.

NRF2 and Bip Interconnection Mediates Resistance to the Organometallic Ruthenium-Cymene Bisdemethoxycurcumin Complex Cytotoxicity in Colon Cancer Cells

Organometallic ruthenium (Ru)(II)-cymene complexes display promising pharmacological properties and might represent alternative therapeutic agents in medical applications. Polyphenols, such as curcumin and curcuminoids, display beneficial properties in medicine, including chemoprevention. Here we analyzed the anticancer effect of a cationic Ruthenium (Ru)(II)-cymene Bisdemethoxycurcumin (Ru-bdcurc) complex. The experimental data show that Ru-bdcurc induced cell death of colon cancer cells in vitro. In response to treatment, cancer cells activated the endoplasmic reticulum (ER)-resident chaperone GRP78/BiP and NRF2, the master regulators of the unfolded protein response (UPR) and the antioxidant response, respectively. Pharmacologic targeting of either NRF2 or BiP potentiated the cytotoxic effect of Ru-bdcurc. We also found that NRF2 and UPR pathways were interconnected as the inhibition of NRF2 reduced BiP protein levels. Mechanistically, the increased Ru-bdcurc-induced cell death, following NRF2 or BiP inhibition, correlated with the upregulation of the UPR apoptotic marker CHOP and with increased H2AX phosphorylation, a marker of DNA damage. The findings reveal that BiP and NRF2 interconnection was a key regulator of colon cancer cells resistance to Ru-bdcurc cytotoxic effect. Targeting that interconnection overcame the protective mechanism and enhanced the antitumor effect of the Ru-bdcurc compound.

Thymoquinone's Potential Role in Cancer Chemotherapy: A Mini Review

Background: Pollution and genetic factors have increased the likelihood of cancer in the developing world. Cancer is now the leading cause of death in young patients. Scientists are focusing on developing new treatments with a lower risk profile than traditional cytotoxic drugs. Nigella sativa (black seed) has been used for centuries by ancient cultures to treat a variety of ailments. Thymoquinone, the main active compound found in Nigella sativa, is a useful therapeutic agent for various morbidities, including cancer. Aim: The review aims to highlight thymoquinone's potential cytotoxic and chemoprotective effects. Methods: The most recent articles were found using reputable websites such as Google Scholar, PubMed, and Research Gate. Many titles, such as thymoquinone, breast cancer, leukemia, colon cancer, osteosarcoma, ovary cancer, and so on, have been used in searches. Then the information from in vitro studies and animal experiments was collected; the preprint, article review, and meta-analysis study were all excluded. Results: Thymoquinone reduced cell viability and induced programmed cell death in breast cancer, colon cancer, leukemia, osteosarcoma, ovary cancer, and colon cancer. TQ causes cytotoxicity through a variety of mechanisms, including the induction of reactive oxygen species and the inhibition of NF-kB activity in some cancers. Conclusion: Thymoquinone is a promising future cytotoxic agent with fewer side effects than traditional cytotoxic agents.

Regulatory Role of Sphingosine-1-Phosphate and C16:0 Ceramide, in Immunogenic Cell Death of Colon Cancer Cells Induced by Bak/Bax-Activation.

We recently identified the sphingosine kinases (SphK1/2) as key intracellular regulators of immunogenic cell death (ICD) in colorectal cancer (CRC) cells. To better understand the mechanism by which SphK inhibition enhances ICD, we focused on the intracellular signaling pathways leading to cell surface exposure of calreticulin (ectoCRT). Herein, we demonstrate that ABT-263 and AZD-5991, inhibitors of Bcl-2/Bcl-XL and Mcl-1, respectively, induce the production of ectoCRT, indicative of ICD. Inhibition of SphK1 significantly enhanced ABT/AZD-induced ectoCRT production, in a caspase 8-dependent manner. Mechanistically, we demonstrate that ABT/AZD-induced Bak/Bax activation stimulates pro-survival SphK1/sphingosine-1-phosphate (S1P) signaling, which attenuates ectoCRT production. Additionally, we identified a regulatory role for ceramide synthase 6 (CerS6)/C16:0 ceramide in transporting of ectoCRT to the cell surface. Together, these results indicate that the sphingolipid metabolic regulators of the sphingolipid rheostat, S1P and C16:0 ceramide, influence survival/death decisions of CRC cells in response to ICD-inducing chemotherapeutic agents. Importantly, SphK1, which produces S1P, is a stress-responsive pro-survival lipid kinase that suppresses ICD. While ceramide, produced by the inhibition of SphK1 is required for production of the cell surface marker of ICD, ectoCRT. Thus, inhibition of SphK1 represents a means to enhance the therapeutic efficacy of ICD-inducing agents.

Melatonin and cisplatin synergistically enhance apoptosis via autophagy-dependent alteration of P53 transcription in human colorectal cancer cells

Cisplatin is one of the most general chemotherapeutic agents used to treat various cancers, including colorectal and breast cancer. because cisplatin has some adverse effects including cardiotoxicity and hepatoxicity, it is usage limited. Melatonin is a natural product that responsible for regulator of circadian rhythms and has anti-cancer potential. However, its synergistic effects with melatonin and cisplatin, its efficacy in cancer cell death, its mechanisms and biological targets are not well understood. In this study, it was aimed to determine the synergistic activity of cisplatin with melatonin in colon cancer cell death through apoptotic and autophagic mechanisms. In the present study, we found that melatonin with cisplatin treatment did not affect the cytotoxicity, but cisplatin increased in 24 h incubation period. Melatonin and combined treatment of melatonin and cisplatin also increased the cytotoxicity in 48h incubation period. It was observed that cisplatin treatments used together with melatonin and melatonin inhibited the mitogen activity of colon cancer cells. In addition, combined treatment of cisplatin and melatonin and single treatment of cisplatin increased both apoptosis and autophagic cell death. The results revealed that the use of melatonin with combined cisplatin has been shown to increase the apoptosis and autophagic cell deaths via P53 gene activation.

Polyphyllin I Promotes Autophagic Cell Death and Apoptosis of Colon Cancer Cells via the ROS-Inhibited AKT/mTOR Pathway

Colon cancer is a common malignant tumor of the digestive tract, and it is considered among the biggest killers. Scientific and reasonable treatments can effectively improve the survival rate of patients if performed in the early stages. Polyphyllin I (PPI), a pennogenyl saponin isolated from Paris polyphylla var. yunnanensis, has exhibited strong anti-cancer activities in previous studies. Here, we report that PPI exhibits a cytotoxic effect on colon cancer cells. PPI suppressed cell viability and induced autophagic cell death in SW480 cells after 12 and 24 h, with the IC50 values 4.9 ± 0.1 μmol/L and 3.5 ± 0.2 μmol/L, respectively. Furthermore, we found PPI induced time-concentration-dependent autophagy and apoptosis in SW480 cells. In addition, down-regulated AKT/mTOR activity was found in PPI-treated SW480 cells. Increased levels of ROS might link to autophagy and apoptosis because reducing the level of ROS by antioxidant N-acetylcysteine (NAC) treatment mitigated PPI-induced autophagy and apoptosis. Although we did not know the molecular mechanism of how PPI induced ROS production, this is the first study to show that PPI induces ROS production and down-regulates the AKT/mTOR pathway, which subsequently promotes the autophagic cell death and apoptosis of colon cancer cells. This present study reports PPI as a potential therapeutic agent for colon cancer and reveals its underlying mechanisms of action.

Non-Canonical Programmed Cell Death in Colon Cancer.

Simple SummaryNon-canonical PCD is an important player in colon cancer cell suicide. It influences colon cancer in many ways, such as through tumorigenesis, treatment, and prognosis. In this review, we present the mechanism, application, and prospect of different types of non-canonical PCD in colon cancer.Programmed cell death (PCD) is an evolutionarily conserved process of cell suicide that is regulated by various genes and the interaction of multiple signal pathways. Non-canonical programmed cell death (PCD) represents different signaling excluding apoptosis. Colon cancer is the third most incident and the fourth most mortal worldwide. Multiple factors such as alcohol, obesity, and genetic and epigenetic alternations contribute to the carcinogenesis of colon cancer. In recent years, emerging evidence has suggested that diverse types of non-canonical programmed cell death are involved in the initiation and development of colon cancer, including mitotic catastrophe, ferroptosis, pyroptosis, necroptosis, parthanatos, oxeiptosis, NETosis, PANoptosis, and entosis. In this review, we summarized the association of different types of non-canonical PCD with tumorigenesis, progression, prevention, treatments, and prognosis of colon cancer. In addition, the prospect of drug-resistant colon cancer therapy related to non-canonical PCD, and the interaction between different types of non-canonical PCD, was systemically reviewed.

Myricetin loaded in solid lipid nanoparticles induces apoptosis in the HT-29 colorectal cancer cells via mitochondrial dysfunction.

Among the flavonoids, Myricetin (MCN) has negligible side effects and anti-cancer properties. However, the therapeutic potential of MCN has been limited mainly by its low bioavailability. Nanocarriers improve the bioavailability and stability of flavonoids. The toxic effects of MCN loaded in solid lipid nanoparticles (MCN-SLNs) on the HT-29 human colorectal cancer cells were investigated in this study. HT-29 cells were exposed to the 30µmol MCN or MCN-SLNs for 24h. Colony formation, cell viability, apoptosis, and expression of the Bax, Bcl-2, and AIF (apoptosis-inducing factor) have been investigated. Mitochondrial membrane potential(MMP) andreactive oxygen species(ROS) generation were also measured. The MCN-SLNs with appropriate characteristics and a slow sustained MCN release until 48h made. MCN-SLNs could diminish colony numbers and survival of the HT-29 cells. The apoptosis index of MCN-SLNs-treated cells significantly increased compared to the free MCN (p < 0.001). The expression of Bax and AIF were elevated (p < 0.01 and p < 0.001, respectively) while Bcl-2 expression was decreased in MCN-SLNs treatment (p < 0.05). Moreover, MCN-SLNs significantly enhanced the ROS formation and reduced MMP compared to the free MCN-treated cells (p < 0.01). The SLN formulation of MCN can effectively induce colon cancer cell death by raising ROS formation and activating the apoptosis process.

6-Gingerol, but Not Whole Ginger Juice, Specifically Inhibits Growth of Colon Cancer Cells in Culture

ObjectivesColon cancer is affluent among many people, and having cancer greatly impacts their lives. Ginger is a common food, particularly in Asian cuisine. However, the health benefits of ginger and 6-gingerol, as its bioactive component in the prevention of colon cancer, have not been fully addressed. This experiment investigated the effects of ginger juice and 6-gingerol on colon cancer cell growth and death. MethodsColon cancer SW480 cells and CCD-18Co normal colon epithelial cells were purchased from ATCC.SW480 cells were grown in DMEM with 4.5 g/L glucose supplemented with 10% fetal bovine serum (FBS), penicillin, and streptomycin, and normal colon CCD-18Co cells were maintained in EMEM with 4.5 g/L glucose and glutamine with 10% FBS. 6-gingerol was dissolved in DMSO in a stock of 100 mmol/L. Ginger roots were homogenized and the juice was collected through 3 layers of cheesecloth filtering, followed by centrifugation and 0.2 μm filter sterilization. About 5000 cells were seeded in a 24-well plate and treated with various amounts of ginger juice and/or 6-gingerol for up to 72 hours. Cell growth was examined using Trypan blue stain, and cell cycle arrest was determined by immunoblotting using antibodies against key proteins in cell cycles. Data were analyzed by two-way ANOVA with a Tukey posthoc test and statistical significance was set at P < 0.05. ResultsTime course and dosage curve experiments showed that 6-gingerol significantly inhibited SW480 cell numbers starting at 0.5 μM (P < 0.005). More than 1 μM 6-gingerol did not give more power to inhibit SW480 cell growth. The results also showed that CCD-18Co cell numbers were not changed after 6-gingerol treatments (P > 0.1). Low dosages of ginger juice (2,000 x- to 100 x- fold dilutions) didn’t affect the growth of both cell types. Immunoblotting results revealed that the elevation of pSer10-CDC2 levels and decreases in p21 Wafl/Cip1 and pSer642-Wee1 only occurred in SW480 but not CCD-18Co cells when treated with 1 μM 6-gingerol for 40 hrs. ConclusionsThrough this experiment, it can be concluded that 6-gingerol can kill SW480 cancer cells without killing normal CCD-18Co cells through cell cycle arrest. Further experiments could be run to discover more properties of 6-gingerol, how it works, and how it could be used in the medical world. Funding SourcesOklahoma State University internal funds to support high school student research.

Colon Cancer Cell Death by Synergistic Activity of Phytochemicals is Modulated by Increased ROS, Activation of AMPK, and Inhibition of mTORC1

Colorectal cancer (CRC) is the second leading cause of cancer‐related death in the United States, and has high prevalence in both men and women. Consumption of foods high in fruits, vegetables, and spices is strongly associated with a reduced risk of developing cancer, and may be attributable to the synergy of phytochemicals present in the diet. Previously, we have shown that combinatorial treatment with curcumin and silibinin B (CS) led synergistically to higher rates of colon cancer cell death in comparis on to single compound treated cells. Our goal is to elucidate the molecular and signaling mechanisms of the synergistically enhanced anticancer activity mediated by CS in CRC. Cancer cells maintain high energy levels by higher rates of glycolysis and lactic acid fermentation, occurring in the abundance of oxygen, and have elevated levels of reactive oxygen species (ROS) compared to normal cells. Cancer cells adapt to increased levels of ROS by developing ROS scavenging mechanisms. Despite this, the ROS levels are higher than in normal cells. Phytochemicals have been shown to increase ROS by interfering with enzymes involved in reducing ROS. Our hypothesis is that phytochemical‐based disruption of ROS scavenging mechanisms increases ROS to higher levels, which leads to the activation of AMPK. Activated AMPK, in turn, inhibits the mTOR pathway thereby increasing cell death by apoptosis and autophagy. Our initial results showed increased ROS and activated AMPK in combinatorial treated cells compared to single compound and control treated cells.

Bioprophylactic potential of novel human colostrum probiotics via apoptotic induction of colon cancer cells and cell immune activation

The potential and modes of action of novel human colostrum probiotics were investigated with the aim of studying their application as an alternative in the bioprophylactic and biotherapy of colon cancer. A total of 218 isolates of Gram-positive rod and cocci bacteria obtained from the colostrum of 50 healthy lactating females were collected. Beneficial probiotic criteria from these isolates, which included growth inhibition against seven foodborne pathogens (Helicobacter pylori, Escherichia coli, Salmonella Enteritidis, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus and Listeria monocytogenes), no blood haemolysis and tolerance to acid and bile salt conditions, resulted in only eight probiotic bacteria successfully inhibiting the proliferation of colon cancer cells at rates of 32.47–61.21%. Two probiotic bacteria with higher anticancer efficacy (Streptococcus salivarius CP163 and S. salivarius CP208) were identified using 16S rRNA sequences. High rates of cell surface hydrophobicity, autoaggregation and coaggregation were obtained from both probiotics. The probiotic mode of action involved synergic probiotic adhesion to colon cancer cells that triggered SCFA bioproduction. Apoptotic induction of colon cancer cell death through caspase-2 activity, DNA fragmentation and morphological change as assessed by AO/PI staining were also observed. Immune stimulation by S. salivarius CP163 and S. salivarius CP208 resulted in B and T-cell lymphocyte activation. This study suggests that these novel human colostrum probiotics could be applied as a functional food to facilitate a bioprophylactic strategy in colon cancer.

The Impact of NRF2 Inhibition on Drug-Induced Colon Cancer Cell Death and p53 Activity: A Pilot Study.

Nuclear factor erythroid 2 (NF-E2) p45-related factor 2 (NRF2) protein is the master regulator of oxidative stress, which is at the basis of various chronic diseases including cancer. Hyperactivation of NRF2 in already established cancers can promote cell proliferation and resistance to therapies, such as in colorectal cancer (CRC), one of the most lethal and prevalent malignancies in industrialized countries with limited patient overall survival due to its escape mechanisms in both chemo- and targeted therapies. In this study, we generated stable NRF2 knockout colon cancer cells (NRF2-Cas9) to investigate the cell response to chemotherapeutic drugs with regard to p53 oncosuppressor, whose inhibition we previously showed to correlate with NRF2 pathway activation. Here, we found that NRF2 activation by sulforaphane (SFN) reduced cisplatin (CDDP)-induced cell death only in NRF2-proficient cells (NRF2-ctr) compared to NRF2-Cas9 cells. Mechanistically, we found that NRF2 activation protected NRF2-ctr cells from the drug-induced DNA damage and the apoptotic function of the unfolded protein response (UPR), in correlation with reduction of p53 activity, effects that were not observed in NRF2-Cas9 cells. Finally, we found that ZnCl2 supplementation rescued the cisplatin cytotoxic effects, as it impaired NRF2 activation, restoring p53 activity. These findings highlight NRF2′s key role in neutralizing the cytotoxic effects of chemotherapeutic drugs in correlation with reduced DNA damage and p53 activity. They also suggest that NRF2 inhibition could be a useful strategy for efficient anticancer chemotherapy and support the use of ZnCl2 to inhibit NRF2 pathway in combination therapies.

Thioredoxin reductase 3 suppression promotes colitis and carcinogenesis via activating pyroptosis and necrosis.

Txnrd3 as selenoprotein plays key roles in antioxidant process and sperm maturation. Inflammatory bowel diseases, such as ulcerative colitis and Crohn's disease, are becoming significantly increasing disease worldwide in recent years which are proved relative to diet, especially selenium intake. In the present study, 8-week-old C57BL/6N male Txnrd3-/-, Txnrd3-/ + , Txnrd3 + / + mice, weight 25-30g, were randomly chosen and each group with 30 mice. Feed 3.5% DSS drinking water and normal water continuously for 7days. Mouse colon cancer cells (CT26) were cultured in vitro to establish Txnrd3 overexpressed/knocked-down model by cell transfection technology. Morphology and ultrastructure, calcium levels, ROS level, cell death were observed and detected in vivo and vitro. In Txnrd3-/-mice, ulcerative colitis was more severe, the morphological and ultrastructural lesions were also more prominent compared with wild-type mice, accompanied by the significantly increased expression of NLRP3, Caspase1, RIPK3, and MLKL. Overexpression of Txnrd3 could lead to increased oxidative stress through intracellular calcium outflow-induced oxidative stress increase followed by necrosis and pyroptosis pathway activation and further inhibit the growth and proliferation of colon cancer cells. Txnrd3 overexpression leads to intracellular calcium outflow and increased ROS, which eventually leads to necrosis and focal death of colon cancer cells, while causing Txnrd3-/- mice depth of the crypt deeper, weakened intestinal secretion and immune function and aggravate the occurrence of ulcerative colitis. The present study lays a foundation for the prevention and treatment of ulcerative colitis and colon carcinoma in clinic 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.