HT-29 Human Colorectal Cancer Cells Research Articles

Moringa oleifera leaf powder – Silver nanoparticles (MOLP-AgNP) efficiently inhibit metastasis and proliferative signaling in HT-29 human colorectal cancer cells

Colorectal cancer is the second most common cancer which caused large number of deaths worldwide. It has been widely known that its undergoing therapies were related to cell metastasis and drug resistance. In line with the endeavor to reduce the side effects of those therapies, bio-fabricated silver nanoparticles (AgNPs), which are synthesized using plant extracts, are massively explored due to their potential as anticancer agents. Specific analytical techniques were used to characterize MOLP-AgNPs, such as UV–visible spectrophotometer (UV–Vis), Particle sizer (PSA), Zeta potential, FT-IR and TEM. HT-29 cells were treated at different single-dose concentrations and combined treatment using Doxorubicin as a well-established cancer drug. Based on the MTT assay result, Moringa oleifera Leaf Powder – Silver Nanoparticles (MOLP-AgNPs) induced significant dose-dependent results according to decreased HT-29 cell viability and proliferation. MOLP-AgNPs reduced cell cycle and proliferation-related genes, such as Ki-67, Wnt, β-catenin, and Cyclin D1 in a dose-dependent manner. Furthermore, MOLP-AgNPs also reduced the expression of metastasis-related genes, such as TGF-β and Snail, known as metastasis inducers. These findings suggest that MOLP-AgNPs could be considered a potential strategy for colon cancer treatment.

Exploration of the aptitude to alleviate oxidative impairment and curb colorectal cancer manifestation by Nostoc calcicola in HT-29 adenocarcinoma cells

BackgroundMarine cyanobacteria have been known to contain several unique bioactive compounds which have different therapeutic potentials. The current research focuses to identify the efficacy of Nostoc calcicola to counteract the harmful effects of free radicals and testing its anticancer activity against colorectal adenocarcinoma cells (HT-29).ResultsMethanol is used as a solvent for the extraction of bioactive metabolites from Nostoc calcicola followed by phytochemical screening representing the presence of flavonoids, phenols, tannins, saponins, and steroids to find out bioactive metabolites. Furthermore, evaluation of the extract efficacy revealed the profound ability of Nostoc calcicola to scavenge free radicals by neutralizing different reactive oxygen species. At 100 µg/mL concentration, it inhibited DPPH radicals (73.4%), enhanced phosphomolybdenum reduction (53.5%), displayed ferric-reducing power (55.1%), and finally the extract revealed remarkable hydroxyl radicals scavenging capacity (94.8%), compared to the standards. These compelling results emphasize the robust antioxidant potential of the Nostoc calcicola extract. In vitro, studies demonstrated the selective cytotoxic effects of methanol extracts of Nostoc calcicola on the HT-29 human colorectal cancer cell line, as indicated by IC50 values of 25 µg/mL for the extracts. Treatment with me Nostoc calcicola decreased the cell viability of HT-29 cells followed by consistent morphological changes leading to cytotoxicity. Nuclear condensation and DNA fragmentation were observed using AO/EtBr and DAPI staining. Flow cytometry analysis further confirmed the incidence of apoptosis during the S phase of the cell cycle. Furthermore, western blotting analysis confirmed the activation of caspase 9, a pivotal enzyme in the intrinsic apoptosis pathway, suggesting the ability of Nostoc calcicola to induce apoptosis in HT-29 colorectal cancer cells.ConclusionThese findings underscore the potential of Nostoc calcicola as a valuable source of bioactive compounds with antioxidant and anticancer properties, warranting further investigation for their potential therapeutic applications in colorectal cancer.Graphical

Piper nigrum Extract Inhibits the Growth of Human Colorectal Cancer HT-29 Cells by Inducing p53-Mediated Apoptosis.

Colorectal cancer (CRC) is a prevalent malignancy of the digestive tract with the second highest mortality rate globally. Piper nigrum is a widely used traditional medicinal plant, exhibiting antitumor activity against various tumor cells. At present, research on the effect of Piper nigrum on CRC is limited to in vitro cytotoxicity, lacking comprehensive mechanism investigations. This study aimed to explore the inhibitory effect and mechanism of Piper nigrum extract (PNE) on HT-29 cells. Firstly, we identified the chemical components of PNE. Then, MTT assay, colony formation assay, JC-1 staining, and flow cytometry were used to analyze the effect of PNE on HT-29 cells in vitro. A xenograft model, histopathological examination, immunohistochemistry, and western blot were used to evaluate the tumor growth inhibitory activity and mechanism of PNE in vivo. The results indicated that PNE could inhibit cell proliferation and colony formation, reduce mitochondrial membrane potential, induce cell apoptosis in vitro, and inhibit tumor growth in vivo. Furthermore, PNE could regulate p53 and its downstream proteins, and subsequently activate the caspase-3 pathway. In summary, PNE probably induced apoptosis of HT-29 cells through the mitochondrial pathway mediated by p53. All these results suggested that PNE might be a potential natural-origin anti-CRC drug candidate.

Sphingosine kinase-2 silencing modulates sensitivity of HT-29 human colorectal cancer cells to paclitaxel-induced cell death through altering apoptosis-related proteins.

The sphingosine kinase-2 (SphK2), a main component of sphingolipid signal transduction, is reported as an innovative therapeutic candidate for cancer treatment. This study was conducted to investigate the suppression of SphK2 in increasing HT-29 colorectal cancer cells sensitivity to paclitaxel via inducing apoptosis and targeting c-FLIPS, MCL-1, and survivin expressions. Cells were transfected with siRNA against SphK2, PTX, or SphK2 activator. Proliferation and apoptosis were evaluated by MTT assay, and trypan blue staining and ELISA, respectively. SphK2, c-FLIPS, MCL-1, and survivin expression were determined by qRT-PCR and Western blotting. SphK2 siRNA increased cell death induced PTX. SphK2 agonist abolished silencing impact on cell survival. We found down-expression of C-FLIPS, MCL-1, and survivin by SphK2 siRNA transfection either alone or in paclitaxel treated cells, as well as elevation in cell apoptosis. Our results showed that SphK2 suppression may be an effective important modality to enhance cell sensitivity to paclitaxel via the induction of apoptosis in colorectal cancers.

Proliferation Inhibitory Activity of Quinones from Blaps rynchopetera Defense Secretion on Colorectal Tumor Cells.

To explore the proliferation inhibitory effect of quinones from Blaps rynchopetera defense secretion on colorectal tumor cell lines. Human colorectal cancer cell HT-29, human colorectal adenocarcinoma cell Caco-2 and normal human colon epithelial cell CCD841 were chosen for the evaluation of inhibitory activity of the main quinones of B. rynchopetera defense secretion, including methyl p-benzoquinone (MBQ), ethyl p-benzoquinone (EBQ), and methyl hydroquinone (MHQ), through methyl thiazolyl tetrazolium assay. The tumor-related factors, cell cycles, related gene expressions and protein levels were detected by enzyme-linked immunosorbent assy, flow cytometry, RT-polymerase chain reaction and Western blot, respectively. MBQ, EBQ, and MHQ could significantly inhibit the proliferation of Caco-2, with half maximal inhibitory concentration (IC50) values of 7.04 ± 0.88, 10.92 ± 0.32, 9.35 ± 0.83, HT-29, with IC50 values of 14.90 ± 2.71, 20.50 ± 6.37, 13.90 ± 1.30, and CCD841, with IC50 values of 11.40 ± 0.68, 7.02 ± 0.44 and 7.83 ± 0.05 µg/mL, respectively. Tested quinones can reduce the expression of tumor-related factors tumor necrosis factor α, interleukin (IL)-10, and IL-6 in HT-29 cells, selectively promote apoptosis, and regulate the cell cycle which can reduce the proportion of cells in the G1 phase and increase the proportion of the S phase. Meanwhile, tested quinones could up-regulate mRNA and protein expression of GSK-3β and APC, while down-regulate that of β-catenin, Frizzled1, c-Myc, and CyclinD1 in the Wnt/β-catenin pathway of HT-29 cells. Quinones from B. rynchopetera defense secretion could inhibit the proliferation of colorectal tumor cells and reduce the expression of related factors, which would be functioned by regulating cell cycle, selectively promoting apoptosis, and affecting Wnt/β-catenin pathway-related mRNA and protein expressions.

Raptinal ameliorates 1,2-dimethylhydrazine-induced colon cancer through p53/Bcl2/Bax/caspase-3-mediated apoptotic events in vitro and in vivo.

Colon carcinoma stands as the most familiar malignancy throughout across the globe. Raptinal induce apoptosis through the alteration of cellular events. Thus, in the present investigation, the anticancer activity of raptinal counter to 1,2-dimethylhydrazine (DMH) persuaded colon carcinoma has been evaluated through both in vivo and in vitro systems. The pharmacophore analysis demonstrated the binding efficacy of raptinal with the apoptotic proteins. The chemotherapeutic activity of raptinal was examined through HT-29 human colorectal cancer (CRC) cell line as well as DMH persuaded CRC in the rat model. The cytotoxicity analysis, flow cytometry, and DAPI analysis have been carried out on HT-29 cell line through in vitro assessment. The colon carcinoma has been induced through DMH administration and subsequently Dextran sulfate sodium treatment in male Wistar rats. After 18 weeks of raptinal treatment, the colon tissues have been investigated for aberrant crypt foci (ACF) count, antioxidant status, histology, immunohistochemical assessment, and apoptotic analysis. The raptinal therapy on HT-29 cells demonstrated a substantial % of early apoptosis followed by G0 and G1 phase arrest, which subsequently led to apoptosis. Furthermore, it inhibits ACF development with improved colonic abrasions and structural integrity of colonic mucosa with increased levels of antioxidants, proapoptotic biomarkers including p53, caspase-3, Bax and downstream effects of Bcl-2, tumor necrosis factor (TNF)-α, and interleukin (IL)-6 mutation. These findings indicate the raptinal effectively reduces colon cancer by inducing apoptosis through p53/Bcl2/Bax/caspase-3 pathway and suppressing IL-6, TNF-mediated chronic inflammation in the colon cancer microenvironment.

Biocatalytic synthesis of oxadiazole thioethers and evaluation of their antitumor activity.

A multicomponent enzyme-catalyzed process is suggested for the synthesis of a novel series of 1,3,4-oxadiazole thioether derivatives with yields ranging from 65 to 94%. Novozym 435, the immobilized form of Candida antarctica lipase B (CALB), was found to efficiently catalyze the reaction. The products were evaluated for antitumor activities against two cancer cell lines, HT-29 (human colorectal cancer cell) and HepG2 (human liver cancer cell), by MTT assays. Among them, two compounds exhibited higher antitumor activities, for both cell lines, compared to doxorubicin. In silico molecular docking and computational ADME analysis were performed to propose a mode of action for the anti-cancer activities and to predict drug-likeness, respectively.

Cedrol, a Sesquiterpene Isolated from Juniperus chinensis, Inhibits Human Colorectal Tumor Growth associated through Downregulation of Minichromosome Maintenance Proteins.

Cedrol, a sesquiterpene alcohol, isolated from Juniperus chinensis has been reported to inhibit minichromosome maintenance (MCM) proteins as cancer biomarkers in human lung cancer in vitro. In the present study, we investigated the anti-cancer activity of cedrol in vitro and in vivo using human colorectal cancer HT29 cells and a human colorectal tumor xenograft model. Cedrol inhibited MCM protein expression and cell growth in HT29 cells, which are associated with G1 arrest and the induction of apoptosis. We demonstrated that cedrol effectively reduced HT29 tumor growth without apparent weight loss in a human tumor xenograft model. Compared with vehicle- and adriamycin-treated tumor tissues, cedrol induced changes in the tumor tissue structure, resulting in a reduced cell density within the tumor parenchyma and reduced vascularization. Moreover, the expression of MCM7, an important subunit of MCM helicase, was significantly suppressed by cedrol in tumor tissue. Collectively, these results suggest that cedrol may act as a potential anti-cancer agent for colorectal cancer by inhibiting MCM protein expression and tumor growth.

Hyperthermia of Magnetically Soft-Soft Core-Shell Ferrite Nanoparticles

Magnetically soft-soft MnFe2O4-Fe3O4 core-shell nanoparticles were synthesized through a seed-mediated method using the organometallic decomposition of metal acetyl acetonates. Two sets of core-shell nanoparticles (S1 and S2) of similar core sizes of 5.0 nm and different shell thicknesses (4.1 nm for S1 and 5.7 nm for S2) were obtained by changing the number of nucleating sites. Magnetic measurements were conducted on the nanoparticles at low and room temperatures to study the shell thickness and temperature dependence of the magnetic properties. Interestingly, both core-shell nanoparticles showed similar saturation magnetization, revealing the ineffective role of the shell thickness. In addition, the coercivity in both samples displayed similar temperature dependencies and magnitudes. Signatures of spin glass (SG) like behavior were observed from the field-cooled temperature-dependent magnetization measurements. It was suggested to be due to interface spin freezing. We observed a slight and non-monotonic temperature-dependent exchange bias in both samples with slightly higher values for S2. The effective magnetic anisotropy constant was calculated to be slightly larger in S2 than that in S1. The magnetothermal efficiency of the chitosan-coated nanoparticles was determined by measuring the specific absorption rate (SAR) under an alternating magnetic field (AMF) at 200-350 G field strengths and frequencies (495.25-167.30 kHz). The S2 nanoparticles displayed larger SAR values than the S1 nanoparticles at all field parameters. A maximum SAR value of 356.5 W/g was obtained for S2 at 495.25 kHz and 350 G for the 1 mg/mL nanoparticle concentration of ferrogel. We attributed this behavior to the larger interface SG regions in S2, which mediated the interaction between the core and shell and thus provided indirect exchange coupling between the core and shell phases. The SAR values of the core-shell nanoparticles roughly agreed with the predictions of the linear response theory. The concentration of the nanoparticles was found to affect heat conversion to a great extent. The in vitro treatment of the MDA-MB-231 human breast cancer cell line and HT-29 human colorectal cancer cell was conducted at selected frequencies and field strengths to evaluate the efficiency of the nanoparticles in killing cancer cells. The cellular cytotoxicity was estimated using flow cytometry and an MTT assay at 0 and 24 h after treatment with the AMF. The cells subjected to a 45 min treatment of the AMF (384.50 kHz and 350 G) showed a remarkable decrease in cell viability. The enhanced SAR values of the core-shell nanoparticles compared to the seeds with the most enhancement in S2 is an indication of the potential for tailoring nanoparticle structures and hence their magnetic properties for effective heat generation.

Sappanin-type homoisoflavonoids from Scilla bisotunensis Speta.: Cytotoxicity, molecular docking, and chemotaxonomic significance

Phytochemical investigation of the chloroform extract of Scilla bisotunensis Speta. bulbs led to the isolation and structure elucidation of six sappinane-4-one (1-4, 7, 9) and three 3,9-dehydrosappinan-4-one (5, 6, 8) homoisoflavonoids. The structures of compounds 1-9 were established based on extensive NMR, ESIMS, UV, and ECD spectra, and also by comparison with the previously reported spectroscopic data. The in vitro cytotoxicity of sappanin-type homoisoflavonoids 1-9 was assayed against HT-29 human colorectal cancer cells using sulforhodamine B assay. Compounds 8 and 9 showed higher cytotoxic effects with IC50 of 5.3 and 6.1 µg/mL, respectively, while compounds 3, 5, and 7 exhibited moderate activity with IC50 ranging from 25 to 37 µg/mL. Interestingly, molecular docking studies revealed that compounds 1‒9 could be potential sarcoma kinase domain inhibitors with binding energies ranging from ‒7.37 (6) to ‒8.34 (9) kcal/mol when compared with the native inhibitor, purvalanol A of ‒7.90 kcal/mol. All homoisoflavonoids, except 1 and 5 are reported for the first time in a member of the Scilla genus, while, to the best of our knowledge this is the first phytochemical and biological study on S. bisotunensis. The plant exhibited good chemotaxonomic relationships to the genus Eucomis L'Hér. on the basis of comparing their chemical constituents with those reported in the related plant families.

Salicylic acid directly binds to ribosomal protein S3 and suppresses CDK4 expression in colorectal cancer cells

Colorectal cancer is a significant cause of morbidity and represents a serious public health issue in many countries. The development of a breakthrough preventive method for colorectal cancer is urgently needed. Aspirin has recently been attracting attention as a cancer preventive drug, and its inhibitory effects on the development of various cancers have been reported in several large prospective studies. However, the underlying molecular mechanisms have not yet been elucidated in detail. In the present study, we attempted to identify the target proteins of aspirin using a chemical biology technique with salicylic acid, the main metabolite of aspirin. We generated salicylic acid-presenting FG beads and purified salicylic acid-binding proteins from human colorectal cancer HT-29 cells. The results obtained showed the potential of ribosomal protein S3 (RPS3) as one of the target proteins of salicylic acid. The depletion of RPS3 by siRNA reduced CDK4 expression and induced G1 phase arrest in human colorectal cancer cells. These results were consistent with the effects induced by the treatment with sodium salicylate, suggesting that salicylic acid negatively regulates the function of RPS3. Collectively, the present results show the potential of RPS3 as a novel target for salicylic acid in the protective effects of aspirin against colorectal cancer, thereby supporting RPS3 as a target molecule for cancer prevention.

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.

Tumor-Inhibitory Effects of Zerumbone Against HT-29 Human Colorectal Cancer Cells.

Colorectal cancer (CRC) is the second cause of cancer-associated death globally. Recently, herbal medicinal products and, in particular, zerumbone have been widely studied and used for cancer treatment as they induce significant anti-cancer effects. However, there is limited information about the anti-cancer effects of zerumbone in CRC. Therefore, we aimed to investigate the in vitro anti-cancer effects of the zerumbone in CRC, focusing on cell apoptosis and migration. Anti-proliferative and anti-migratory effects of zerumbone on HT-29 cells were evaluated using MTT and scratch wound healing assay, respectively. Quantitative real-time PCR (qRT-PCR) was performed to determine the mRNA expression levels of migration and apoptosis-related genes. Apoptosis and cell cycle distribution were evaluated by flow cytometry. The intracellular level of reactive oxygen species (ROS) was measured using a ROS assay kit. Additionally, matrix metalloproteinase-2/-9 (MMP-2/-9) activity was determined using gelatin zymography. Zerumbone suppressed the viability of the HT-29 cells dose-dependently while having less cytotoxicity on normal NIH/3T3 cells. Zerumbone induced apoptosis in HT-29 cells and arrested the cell cycle in the G2/M phase. These effects were associated with alteration in the expression of apoptosis-related genes (up-regulation of Bax and down-regulation of Bcl-2 genes). Zerumbone also enhanced the generation of ROS in HT-29 cells. Furthermore, zerumbone significantly inhibited the migration of HT-29 cells and decreased MMP-2/-9 mRNA expression and activity. Our findings provide a potential use for zerumbone to induce apoptosis and suppress metastasis in HT-29 cells; thus, it could be developed as a promising natural agent for future CRC therapy.

Abstract 5475: DNA methylation regulator cordycepin suppresses gastric and colorectal cancer in combination with chemotherapy

Abstract Gastric cancer and colorectal cancer are the fifth and third most frequent malignant tumors, respectively. One of the most prevalent epigenetic characteristics in gastric and colorectal cancer is aberrant hypermethylation in the promoter of numerous tumor suppressor genes. Hypermethylation of the promoter inactivates tumor suppressor expression and contributes to tumor progression. DNA methyltransferases (DNMTs), which mediate DNA hypermethylation, are putative drug targets of cancer therapy. Several DNMTs inhibitors are under development. 5’-azacytidine and decitabine have been approved for the treatment of AML, myelodysplastic syndrome, and CMML. Clinical trials investigating DNMTs inhibitors in combination with other drugs are ongoing. However, clinical use of 5’-azacytidine and decitabine has been limited due to their toxicity. We have ever demonstrated that cordycepin, a major ingredient of Chinese traditional medicine Cordyceps, has activity of DNMTs inhibition. Treating cancer cells reversed hypermethylation of some tumor suppressor genes promoter, reactivated gene expression, and inhibited cancer cell proliferation. Here, we report the preclinical anti-cancer efficacy of Cordycepin in colorectal and gastric cancers in combination with chemotherapy drugs. In SGC-7901 human gastric cancer cells derived nude mice xenograft tumor model, mice were dosed 20mg/kg Cordycepin alone, 400mg/kg Capecitabine alone, or in combination orally, once daily, for 14 days. Tumor Growth Inhibition rate (TGI) of 89.2% was achieved in the combination therapy group comparing to 57.8% in the 400mg/kg Capecitabine mono-therapy group, and less than 10% in the Cordycepin alone group. Additionally, 20mg/kg Cordycepin plus 3mg/kg fluorouracil achieved TGI of 81.3% after 28 days of treatment in this model, comparing to 67.5% in the 6mg/kg fluorouracil monotherapy group. In HT-29 human colorectal cancer cells derived xenograft model, 20mg/kg Cordycepin plus 250mg/kg Capecitabine orally dosed once daily for 14 days showed TGI of 72.6%, similar to that treated by 400mg/kg Capecitabine alone (80.1%). 20mg/kg Cordycepin plus 2.5mg/kg Tegafur/Gimeracil/Oteracil once daily for 14 days had TGI of 76.7%, while 5mg/kg Tegafur/Gimeracil/Oteracil alone treatment group had TGI of 66.1%. 20mg/kg Cordycepin alone treatment showed only weak inhibition of tumor growth. Synergistic effects of Cordycepin and chemotherapy drugs were observed in treating both tested tumor models. Better tolerance was also shown in the combination therapy groups, as indicated by the mice body weight change. It is demonstrated that the DNMTs regulator Cordycepin has improved the anti-cancer activity and tolerance of chemotherapy drugs, such as Capecitabine, 5-FU, Tegafur/Gimeracil/Oteracil. Clinical development of Cordycepin and chemotherapy combination for cancer patients is strongly supported. Citation Format: Qing Dong, Yangze Cao, Jianyu Liu, Yang Qin. DNA methylation regulator cordycepin suppresses gastric and colorectal cancer in combination with chemotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5475.

Ficus dubia Latex Extract Induces Cell Cycle Arrest and Apoptosis by Regulating the NF-κB Pathway in Inflammatory Human Colorectal Cancer Cell Lines.

Simple SummaryIn this study, the anti-tumor activity of Ficus dubia latex extract (FDLE) against HCT-116 and HT-29 human colorectal cancer cell lines in normal and inflammatory conditions was investigated. The results indicated that an inflammation-activated NF-κB increased proliferation-related proteins and decreased apoptosis-related proteins, leading to hyperproliferation of colorectal cancer cell lines. FDLE exhibited remarkable antiproliferative activity in both conditions, however, more effective anti-proliferation was observed due to its more effective regulation of NF-κB inactivation and some proteins related to cell cycle progression and apoptosis induction in inflammatory condition. Our finding may promote the antiproliferative study of FDLE in inflammation-induced colorectal cancer progression.Colorectal cancer is one of the most diagnosed cancers that is associated with inflammation. Ficus dubia latex is recognized as a remedy with various therapeutic effects in traditional medicine, including anti-inflammatory and antioxidant activity. The present study aims to compare the anti-tumor activity of Ficus dubia latex extract (FDLE) against HCT-116 and HT-29 human colorectal cancer cell lines in normal and inflammatory condition and explore its mechanism of action. FDLE exhibited remarkable antiproliferative activity against HCT-116 and HT-29 colorectal cancer cell lines in both conditions using MTT and colony formation assays and more effective anti-proliferation was observed in inflammatory condition. Mechanistically, FDLE induced cell cycle arrest at G0/G1 phase by down-regulating NF-κB, cyclin D1, CDK4 and up-regulatingp21 in both cell in normal condition. In inflammatory condition, FDLE not only exhibited stronger induction of cell cycle arrest in both cells by down-regulating NF-κB, cyclin D1, CDK4 and down-regulating p21, but also selectively induced apoptosis in HCT-116 cells by down-regulating NF-κB and Bcl-xl and up-regulating Bid, Bak, cleaved caspase-7 and caspase-3 through stronger ability to regulate these proteins. Our results demonstrated that the phytochemical agent in the latex of Ficus dubia could potential be used for treatment and prevention of human colorectal cancer, especially in inflammation-induced hyperproliferation progression.

Effects of Taheri Consciousness Fields on the HT29 Human Colorectal Cancer Cells

Colorectal cancer accounts for 11% of all cancers diagnosed, and it is the second deadliest cancer worldwide. Surgery and chemotherapy or targeted therapy are generally used for the treatment of colorectal cancer. However, there are significant challenges, such as recurrence of tumor and drug resistance, so the application of novel methods is required for the treatment of this cancer. Taheri Consciousness Fields (TCFs) were founded and introduced by Mohammad Ali Taheri. These new fields are not energy or matter and cannot be measured directly. However, we can evaluate the effects of TCFs indirectly through various kinds of research in the laboratory. This study aimed to evaluate the effect of TCFs (A and B) on the HT29 human colon cancer cells in two and three announcements compared with the control group. The morphology and microscopic properties of cells were investigated in TCF (A) and TCF (B) groups compared with the control, and the cells were detached and stained with trypan blue, then the dead cells were counted. To evaluate the inhibitory effect of the TCFs, the MTT assay was used. The expression of two apoptosis-related genes (Bax and Bcl-2) was assessed using RT Real-time PCR. The results demonstrated that TCFs decreased the cell number and changed the morphology of the HT29 cells. In trypan blue dye count, more dead cells were counted in the TCFs groups compared with the control. In the MTT assay, both TCFs decreased the viability of cells on HT29 cells during incubation times (p&lt; 0.05). Bax/Bcl2 ratio increased 4.9 and 7.6-fold in the TCF (A) and TCF (B) treated cells, respectively (P&lt;0.05). Therefore, TCF (A) and TCF (B) induced apoptosis in HT29 cells. The TCF (B) effect was greater than the TCF (A) effect in all tests. There are still many ambiguities and questions about the nature and effects of TCFs. To clarify the issue, more research is needed in vitro, in vivo, and clinically.

MRI assessment of glutamine uptake correlates with the distribution of glutamine transporters and cancer stem cell markers

Glutamine provides carbon and nitrogen for macromolecular synthesis and participates in adenosine triphosphate (ATP) generation, anabolic metabolism, redox homeostasis, cell signaling, and cancer stem cell (CSC) metabolism. New treatment strategies targeting glutamine metabolism in cancer have emerged recently. We previously reported the magnetic resonance imaging (MRI) assessment of glutamine uptake by tumors and activated glutamine metabolism in CSC. In the present study, using MRI, we determined the correlation between glutamine uptake and the distribution of glutamine transporters, namely ASCT2 and SLC38A2 (SNAT2), glutaminase (GLS), and CSC markers, such as CD44 and CD166, in a mouse xenograft model of HT29 human colorectal cancer cells. MRI data revealed an obvious change in intensity following glutamine administration. Immunohistochemistry (IHC) results indicated that ASCT2 staining was stronger in regions that exhibited high glutamine uptake (p = 0.0079). Significant differences were found in the IHC staining intensities of SNAT2, GLS, and CSC markers in the areas of high and low glutamine uptake (p = 0.0079, p = 0.0159 and p = 0.0079, respectively). We also investigated the effect of an ASCT2 inhibitor on the uptake of glutamine using MRI. A statistically significant difference in the initial glutamine uptake was found after ASCT2 inhibitor administration. To conclude, glutamine uptake is positively correlated with the distribution of ASCT2 and certain CSC markers.

Suppressive GLI2 fragment enhances liver metastasis in colorectal cancer.

Linoleic acid (LA) has been shown to cause inflammation and promote development of colorectal cancer (CRC). Moreover, many literatures show that LA is associated with cancer metastasis. Metastatic cancer cells have high stemness, suggesting that LA might affect the stemness of cancer cells. In this study, we examined the effect of LA on the hedgehog system, which affects cancer stemness. In CT26 cells, LA treatment induced the expression of sonic hedgehog (Shh); the signal transduction factor, and glioma-associated oncogene homolog (Gli) 2, whereas the expression of SRY-box transcription factor (Sox) 17 was suppressed. Furthermore, LA reduced GLI2 ubiquitination, resulting in an increase in the N-terminal fragment of GLI2, known as suppressive GLI2, produced by cleavage of GLI2. LA-induced cleaved GLI2 was also detected in Colo320 and HT29 human CRC cells. Knocking down Gli2 abrogated the LA-mediated suppression of Sox17 expression. These results suggest that LA promotes tumor cell stemness by increasing of suppressive GLI2 fragments via GLI2 modification. In mouse liver metastasis models, LA enhanced metastasis with production of the suppressive GLI2 fragments in CT26 and HT29 cells, whereas knockdown of GLI2 abrogated LA-induced metastatic activity. In human CRCs, the cases with liver metastasis showed the suppressive GLI2 fragments. This study provides mechanistic insights into LA-induced stemness in colon cancer cells. This finding suggests that dietary intake of LA might increase the stemness of cancer cells and enhance metastatic activity of the cancer.

Granatin B and punicalagin from Chinese herbal medicine pomegranate peels elicit reactive oxygen species-mediated apoptosis and cell cycle arrest in colorectal cancer cells

BackgroundColorectal cancer ranks among the most common cancers. 5-Fluorouracil (5-FU) based first-line chemotherapy for colorectal cancer treatment often leads to chemoresistance and gastrointestinal mucositis. PurposeThis study aimed to find potential therapeutic agents from herbal medicine with anti-colorectal cancer and anti-mucositis activities. MethodsChinese medicine theory, network pharmacology analyses, and antioxidant activity coupled with liquid chromatography tandem mass spectrometry analyses were used to identify potential bioactive compounds. HT-29 human colorectal cancer cell culture and xenograft tumor models were employed to study anti-colorectal cancer efficacy. Lipopolysaccharide-induced RAW 264.7 and 5-FU treated Dark Agouti rats were used to evaluate anti-inflammatory and anti-mucositis activities. Histological staining, immunofluorescence imaging, western blots, and flow cytometric analyses were employed to explore the underlying mechanisms. ResultsBoth Chinese medicine theory and network pharmacology analyses indicated pomegranate peels as a potential anti-colorectal cancer and anti-mucositis agent. Antioxidant activity coupled with liquid chromatography tandem mass spectrometry analyses revealed granatin B and punicalagin as the most potent antioxidant compounds in pomegranate peels. Granatin B and punicalagin demonstrated superior anti-colorectal cancer activities in both cell culture and xenograft tumor models. Granatin B and punicalagin also exhibited strong anti-inflammatory activities in lipopolysaccharide-induced RAW264.7 cells and anti-mucositis activities in 5-FU-treated rats. Mechanistic studies revealed that granatin B and punicalagin induced reactive oxygen species-mediated S-phase cell cycle arrest and apoptosis in HT-29 cells. Moreover, these compounds sensitized HT-29 cells to 5-FU-induced cell death and S-phase cell cycle arrest. ConclusionWe report that granatin B and punicalagin exhibit superior anti-colorectal cancer and anti-mucositis activities. To the best of our knowledge, these results are novel and suggest that utilizing phenols from herbal medicine, such as granatin B and punicalagin, to target reactive oxygen species may be an innovative therapy to treat colorectal cancer and intestinal mucositis.

Effects of octacosanol on HMG-CoA reductase and cyclooxygenase-2 activities in the HT-29 human colorectal cancer cell line

Effects of octacosanol on HMG-CoA reductase and cyclooxygenase-2 activities in the HT-29 human colorectal cancer cell line