Colorectal HCT116 Research Articles

Unlocking the therapeutic potential of Saussurea costus: purification and functional characterization of α-amylase inhibitors

IntroductionRegulating the catalytic activity of alpha-Amylase enzymes can decrease glucose production during the postprandial phase, potentially offering therapeutic benefits for diabetes. This research aimed to assess the inhibition of α-amylase using crude extracts from Saussurea costus, a medicinal plant traditionally used for treating diabetes and its associated complications.MethodsTwo novel potent proteinaceous amylase inhibitors: ScAI-R and ScAI-L were purified and characterized from Saussurea costus roots and leaves.ResultsThe pure inhibitors exhibited an apparent molecular weight of about 16 kDa and a high N-terminal sequence identity (81%) with the monomeric α-amylase inhibitors from Kengyili amelanthera and Triticum dicoccoides. In addition to their significant stability at extreme pH values (2.0–12.0) and temperatures (100°C), the structural integrity of both inhibitors was remarkably enhanced in the presence of divalent cations such as Mg2+, Ca2+, and Hg2+ at 5 mM. Interestingly, the half-maximal inhibitory concentrations of ScAI-R (IC50 = 23 μg/mL) or ScAI-L (IC50 = 28 μg/mL) against human salivary amylase against were comparable to that of the standard drug acarbose (IC50 = 23 μg/mL). Both purified inhibitors acted as non-competitive inhibitors with Ki values of 0.38 and 0.32 µM, respectively, and displayed the highest affinities towards human salivary and pancreatic α-amylases (up to 90% inhibitory activity) and, to a lesser extent, porcine pancreatic α-amylase (∼70% inhibitory activity). Furthermore, these inhibitors exhibited efficient antimicrobial activities against Gram (−) and Gram (+) bacteria, as well as fungal strains. Cytotoxicity towards the human cancer colorectal cells LoVo and HCT-116 with an IC50 of up to 50 μg/mL was also observed.DiscussionThus, Saussurea costus α-amylase inhibitors could be potential candidates for hyperglycemic control in diabetic and colorectal cancer patients.

Development of 1,2,3-triazole hybrids as multi-faced anticancer agents co-targeting EGFR/mTOR pathway and tubulin depolymerization.

Novel 1,2,3-triazole hybrids bearing various substituents have been synthesized as potential anticancer agents. Ligand-based approach has been adopted to design these compounds relying on the hybridization of 1,2,3-triazole with α,β-unsaturated carbonyl, 5- and 6-membered heterocyclic scaffolds. All synthesized members were investigated for their cytotoxic potency against nine types comprising 60 panels of human cancerous cells by the US National Cancer Institute: Development Therapeutic Program (US_NCI_DTP). Among the tested members, 4b, 4e, and 4h showed prominent cytotoxic effects (> 80% growth inhibition: GI) on a wide panel of tested cancer cell lines, mainly melanoma and colorectal cancer redeeming their selection for five dose testing. Presenting low nanomolar GI50 concentrations, two representative potent anticancer compounds 4b and 4e were subjected to cytotoxicity testing on colon normal cell (FHC) to investigate their safety window and they showed less toxicity to normal cells at the concentration required to produce anticancer effect. Furthermore, 4b and 4e were exposed to additional mechanistic studies in colorectal cancer cell HCT-116 suggesting multifaceted mechanisms of action. A study into the effects of cytotoxic chemicals 4b and 4e on cell cycle progression regulation showed triggered the arrest of cell cycles during the G1 and S phases. Moreover, 4b and 4e caused cell death mainly through apoptosis the thing that has been reinforced by the elevated Bax: Bcl2 ratio, as well as concentrations of caspases 3 and 9 within HCT-116. Further, both compounds showed prominent inhibition profiles against tubulin polymerization as well as EGFR catalytic activity reaching down to low-digit micromolar and sub-micromolar concentrations, respectively, as compared to positive reference controls. Compounds' impacts on gene expression of cancer-associated and EGFR-downstream signaling markers including TNFα, IL-6, and mTOR, were explored in HCT-116 highlighted significant downregulations versus the untreated cells. Docking studies demonstrated the specific fit of 4b and 4e into EGFR and the colchicine binding site of tubulin.

Characterization of TFIIE-regulated genes by transcriptome analysis.

Previous studies on general transcription factor II E (GTF2E) showed that it is associated with certain groups of diseases, such as colon cancer and trichothiodystrophy, but the global effect of GTF2E on cellular processes is still not widely characterized. This study aimed to investigate and characterize the effect of GTF2E on the transcription level of genes and identify the cellular processes and diseases associated with GTF2E. The human colorectal carcinoma cell line HCT116 used in the study was transfected at a 30 nM concentration with siGTF2E1 or nontarget negative siRNA. After 72 h, cells were harvested and prepared for further analysis. A whole transcriptome analysis was performed on the HCT116 cell line obtained from the siGTF2E1 knockdown of the HCT116 cells (n = 3) and their nontarget negative siRNA controls (n = 3) using RNA sequencing. Cell viability was tested using an MTS assay. Compared with the control group, 166 genes were identified at the time of the GTF2E1 knockdown and expressed differentially in the knockdown group, including 66 upregulated genes and 100 downregulated genes. One significantly enriched Gene Ontology term was identified, involving carbohydrate binding. One oncogene related to B cell chronic lymphocytic leukemia (B-CLL) was identified. Five genes associated with colon carcinoma were determined. Eleven genes involved in the development of atherosclerosis were identified. GTF2E1 knockdown caused a decrease in cell viability. The GTF2E1 knockdown group exhibited an altered expression of multiple genes, some of which are related to the development of atherosclerosis, colon carcinoma, and B-CLL. This might shed light on the different regulatory effects of GTF2E and its association with certain diseases.

Creation of a Model Line of Tumor Cells with Inducable Expression of Adenoviral E1A to Study Its Antiproliferative and Cytotoxic Properties In Vitro and In Vivo

Over the past decades, gene therapy based on the adenoviral E1A has proven its benefit against a number of tumor diseases, both in animal models and in clinical studies. It has been shown that in addition to its own antiproliferative activity, E1A also has the ability to enhance the cytotoxic effect of some anticancer drugs. The use of E1A in combination therapy can solve a number of problems in clinical oncology, among which the most pressing is the problem of drug-resistance of tumor cells. This work describes the establishment of a cell model based on human colorectal cancer cells HCT116 and cisplatin-resistant HCT116/C cells with doxycycline-inducible expression of adenoviral E1A. We have shown the concentration-dependent and time-dependent dynamics of E1A expression upon doxycycline treatment, and shown the antiproliferative effect of adenoviral E1A in the HCT116-E1A and HCT116/C-E1A cells in vitro in experiments assessing viability in MTT and clonogenic activity tests and in vivo in xenograft mouse models. Thus, as a result of our work, a model was created to explore the antiproliferative and sensitizing properties of E1A in platinum-sensitive and platinum-resistant colorectal cancer cells and to search for new approaches to anticancer therapy both in vitro and in vivo. The resulting cell line is a convenient model for selecting the most promising combinations of cytostatic drugs with E1A-based gene therapy.

Network-Pharmacology-Based Study on Active Phytochemicals and Molecular Mechanism of Cnidii Fructus in Treating Colorectal Cancer.

Cnidii Fructus (CF) is known for its antibacterial, anti-inflammatory, and antitumor properties, as well as its activities against kidney deficiency and impotence. In this study, we aimed to explore the anti-CRC cancer effect and molecular mechanism of CF via network pharmacology and in vitro antitumor experiments. Network pharmacology was used to investigate the anti-CRC mechanism of CF. First, a series of databases was used to screen the active phytochemical targets and anti-CRC core targets. Then, the GO and KEGG pathways were analyzed to predict possible mechanisms. Molecular docking analysis explore core targets-phytochemicals interactions. In vitro antitumor experiments were carried on verifying anti-CRC mechanism of CF. In this study, 20 active ingredient targets and 50 intersecting targets were analyzed by Cytoscape software 3.9.1 to obtain the core genes and phytochemicals. Then, the GO and KEGG pathways of 50 intersecting targets were analyzed to predict possible mechanisms. The results from GO and KEGG indicated that CF has significant antitumor efficacy, which involves many signaling pathways, such as PI3K/AKT and p53. The five core targets and five core phytochemicals were screened for molecular docking to show protein-ligand interactions. According to the results of molecular docking, the compound O-acetylcolumbianetin was selected for the anti- CRC functional verification in vitro. MTT assay showed that O-acetylcolumbianetin significantly inhibited the proliferation of colorectal HCT116 cells in a time- and quantity-dependent manner. O-acetylcolumbianetin can promote the expression of CASP3 protein, induce HCT116 cells apoptosis, thus exert anti-CRC effect. This study preliminarily verified the anti-CRC effect and molecular mechanism of CF and provided a reference for Traditional Chinese Medicine anti-tumor subsequent research.

Determining the anticancer effect of Gefitinib-loaded NLCs through investigating the inflammatory cytokine (JAK-STAT) pathway

This research aims to investigate the feasibility of using nanostructured lipid carriers (NLC) loaded with Gefitinib to treat colorectal cancer by focusing on the inflammatory cytokine pathway, particularly on the JAK-STAT pathway. As part of the study, Gefitinib-loaded NLC (Nano-GEF) was prepared by homogenization method. Colorectal cancer cells HCT116 were subjected to a series of tests to determine cytotoxicity, including morphological assessment, MTT assay, DNA fragmentation analysis, evaluation of interleukin cytokines, and JAK-STAT pathway expression. According to the study's results, Nano-GEF has far stronger anti-cancer effects than the pure drug (GEF) against cells of colorectal cancer. Morphological studies and cytotoxicity assays confirm the selective action of Nano-GEF on cancer cells. Furthermore, the expression of pathways mediated by JAK-STAT provides further evidence of Nano-GEF's effectiveness as a potential anti-cancer therapy. By focusing on the inflammatory cytokine pathway (JAK-STAT), this research shows that gefitinib-loaded NLC might be a good technique to treat colorectal cancer. Based on the results, Nano-GEF has the potential to be an effective anti-cancer method that is more selective than the pure drug. These findings open the door to the creation of innovative pharmaceutical products as well as the possibility of applications in the assessment and treatment of cancer.

Exploring the therapeutic potential of ruthenium(II)-bis(benzimidazol-2-yl)pyridine complex on MDA-MB-231, HCT-116 and normal L6 cell lines

The in vitro anticancer efficacy and cytotoxicity of the [Ru(bbp)2]2+ complex (bbp = 2,6-bis(benzimidazol-2-yl) pyridine) on human breast cancerous MDA-MB-231, colorectal HCT-116, and normal cell lines were investigated by direct microscopic and MTT assay methods. The synthesised [Ru(bbp)2]2+ complex was characterised by UV–visible, FTIR, 1H NMR, and MALDI-TOF-MS spectroscopic techniques. The complex showed a metal-to-ligand charge transfer (MLCT) absorption peak at 481 nm. The FTIR spectrum of the complex showed absorption bands in the range of 3458–573 cm−1. The 1H NMR chemical shift values resembled those of the bbp ligand. The molecular ion peak (M+) of the [Ru(bbp)2]2+ complex was obtained at m/z 1013.48. The IC50 values of the complex on MDA-MB-231, HCT-116 and normal L6 cell lines were calculated from the percentage cellular viability and are found to be 28.71, 9.06 and 153.82 µM respectively. The morphological changes in the cell lines at various concentrations of the complex were attributed to the generation of reactive oxygen species (ROS). The obtained result revealed that the anticancer activity of the synthesised complex depends on the concentration of the complex. The complex reduced the proportion of viable cells in both malignant cell lines, causing apoptosis. The results suggeseted that the synthesised complex showed good anticancer activity on the MDA-MB-231 and HCT-116 cell lines with low cytotoxicity. Thus, the present investigation paved the way for the development of novel anticancer drugs.

Design and Development of [1,2,4]Triazolo[4,3-b][1,2,4]triazines as Potential Anticancer Agents with Genotoxicity and Apoptotic Activity

Aims: In this current study, a new series of triazolo-triazine derivatives were designed and synthesized as potential anticancer agents. Methods: The antiproliferative activity of the new compounds was evaluated against three different cancerous cell lines (MDA-MB-231, HCT-116, A549, and HT-29) using an MTT assay. To evaluate the mechanism of action, the ability of the best compound in apoptosis induction and DNA damage was evaluated using the flow cytometry technique and comet assays. Furthermore, molecular docking simulation was used to investigate their interactions with the two targets, VEGFR2 and c-Met kinases. Results: Results showed that 6-(4- bromophenyl)-3-((4-methoxybenzyl)thio)-[1,2,4]triazolo[4,3-b][1,2,4]triazine (8c) demonstrated the best anti-proliferative activity against the human colorectal carcinoma cells HCT-116 with an IC50 value of 38.7 ± 1.7 μM. In silico evaluations showed that the triazolo-triazine scaffold, along with the methoxy substitution of compound 8c, was involved in creating effective H-bond interactions in the active site of both targets. Conclusion: Our results showed that compound 8c significantly increased cell death through apoptosis induction and caused a significant increase in genotoxicity. Furthermore, it was found that the tested compound 8c, with a selectivity index of 1.74, possessed selective antiproliferative activity towards the colorectal cancer cell line HCT-116 compared to the normal fibroblast cell line. These findings could be useful in the development of novel VEGFR2/c-Met dual-targeted inhibitors in the future.

Uncovering the Potential of Chalcone-Sulfonamide Hybrids: A Systematic Review on Their Anticancer Activity and Mechanisms of Action.

Cancer is the second leading cause of death worldwide and is considered a major public health problem. Despite the significant advances in cancer research, the conventional cancer treatment approaches often lead to serious side effects that affect the quality of life of cancer patients. Thus, searching for new alternatives for cancer treatment is crucial to minimize these problems. Chalcone-sulfonamide hybrids display a range of biological activities and have been widely investigated for their anticancer potential, being considered promising molecules for cancer treatment. This systematic review aimed to summarize the information available in the literature about the anticancer potential of chalcones-sulfonamides in vitro and in vivo and their mechanisms of action. Our analysis demonstrated that chalcones-sulfonamides have relevant cytotoxic potential against different cancer cell lines in vitro, especially against the human colorectal carcinoma cell line HCT-116. These molecules have also reduced tumor growth in vivo. Some chalcones-sulfonamides had improved cytotoxicity after chemical modification and could become more selective or even more potent than reference chemotherapeutics. The mechanisms underlying these effects demonstrated that chalcones-sulfonamides may lead to cell death by different pathways, predominantly via apoptosis or necroptosis. This review may encourage researchers to advance studies with chalcones-sulfonamides, especially to elucidate their mechanisms of action, contributing to the development of new alternatives to cancer treatment.

Colorectal cancer cell-derived exosomal miRNA-372-5p induces immune escape from colorectal cancer via PTEN/AKT/NF-κB/PD-L1 pathway

Tumor cells can escape immune surveillance by changing their own escape or expressing abnormal genes and proteins, resulting in unlimited proliferation and invasive growth of cells. These changes are related to microRNAs (miRNAs), which reduce the killing effect of immune cells, devastate the immune response, and interfere with apoptosis through the aberrant expression of relevant miRNAs. In the preliminary phase of this study, miRNAs in clinical plasma exosomes of colorectal cancer patients were differentially analyzed by RNA sequencing technology, and miR-372-5p derived from extracellular vesicles (sEVs) was found to be a key signaling molecule mediating the regulation of macrophages by colorectal cancer (CRC). miRNA-372-5p is upregulated in colorectal cancer patient tissues and serum, as well as colorectal cancer cell lines and their exosomes. Subsequently, we found that macrophages could take up sEV secreted by colorectal cancer cells HCT116, affecting the expression of the immune checkpoint PD-L1, resulting in the generation of a tumor-immunosuppressive microenvironment and suppression of T cell activation in CRC. Gene enrichment mapping and database revealed that miR-372-5p regulates PD-L1 expression in colorectal cancer through the homologous phosphatase-tensin (PTEN)-phosphatidylinositol 3-kinase-protein kinase B (AKT)-nuclear factor-κB (NF-κB) pathway. Further studies confirmed that miRNA-372-5p-treated macrophages co-cultured with T cells affected the regulation of PD-L1 expression through the PTEN/AKT/NF-κB signaling pathway, resulting in decreased CD3+CD8+ T cell activity, decreased cytokine IL-2 and increased IFN-γ. And miRNA-372-5p could down-regulate the expression of PD-L1 in HCT116 through the PTEN/AKT/NF-κB pathway, inhibit tumor cell proliferation and promote apoptosis. Conclusion: Colorectal cancer cell-derived exosome miR-372-5p can be phagocytosed by colorectal cancer and macrophage cells, regulate the expression of PD-L1 in colorectal cancer cells and macrophages by targeting the PTEN/AKT/NF-κB pathway, and induce the immunosuppressive microenvironment of CRC to promote CRC development. This suggests that inhibiting the secretion of HCT116-specific sEV-miR-372-5p or targeting PD-L1 in tumor-associated macrophages could be a novel approach for CRC treatment and possibly a sensitizing approach for CRC anti-PD-L1 therapy.

Construction of anticancer drug incorporated aptamer-functionalized cationic β-lactoglobulin: induction of cell cycle arrest and apoptosis in colorectal cancer

Nanoscale drug delivery systems that are both multifunctional and targeted have been developed using proteins as a basis, thanks to their attractive biomacromolecule properties. A novel nanocarrier, aptamer (AS1411)-conjugated β-lactoglobulin/poly-l-lysine (BLG/Ap/PL) nanoparticles, was developed in this study. To this unique formulation, the as-prepared nanocarrier blends the distinctive features of an aptamer as a chemotherapeutic targeting agent with those of protein nanocarriers. By loading cabazitaxel (CTX) onto the nanocarriers, the therapeutic potential of BLG/Ap/PL could be demonstrated. The CTX-loaded BLG/Ap/PL (CTX@BLG/Ap/PL) showed a regulated drug release profile in an acidic milieu, which could improve therapeutic efficacy in cancer cells and have a high drug encapsulation efficacy of up to 93%. However, compared to free CTX, CTX@BLG/Ap/PL killed colorectal HCT116 cancer cells with a higher efficacy at 24 and 48 h. Further investigation confirms the apoptosis by acridine orange and ethidium bromide (AO/EB), and DAPI staining confirms the morphological changes, chromatin condensation, and membrane blebbing in the treated cell through flow cytometry displayed the release of higher percentages of apoptosis. Cell cycle analysis revealed that CTX@BLG/Ap/PL induced sub-G1 and G2/M phase (apoptosis) at 24 and 48 h. Annexin V/propidium iodide (PI) flow cytometry analysis confirmed that CTX@BLG/Ap/PL induces apoptosis in HCT116 cells. Overall, this study proved that CTX@BLG/Ap/PL had several advantages over free chemotherapeutic drugs and showed promise as a solution to the clinical problems associated with targeted antitumor drug delivery systems.

Microwave synthesis, density functional theory study and antiproliferative activity of the novel spiropyrazole derivatives

Different spiro-pyrazole-oxy-indoline substituted compounds I-IV have been synthesized. In Jaguar, Hybrid DFT using Becke’s three-parameter transfer possibility also,Lee-Yang-Parr correlated practical (B3LYP) viewwith a 6-31 G** basis set were utilized to improve the geometrical characteristics of the hit molecules and reference compound. The MTT test exhibited whether the prepared substance has an inhibitory impact on cell development or not. This was successfully represented that in hepatocellular carcinoma (HEPG-2), colorectal carcinoma (HCT-116), and mammary gland breast cancer cell lines (MCF-7) colorimetric test, the mitochondrial succinate dehydrogenase in live cells transform yellow tetrazolium bromide (MTT) to a purple formazan product. Compound IV was found to be a better cytotoxic action against to the HEPG-2 hepatocellular carcinoma cell line, and it has been seen a strong activity against the colorectal carcinoma cell lines HCT-116 and MCF-7 mammary gland breast cancer cell lines. While the other three compounds I, II, and III showed a moderate cytotoxic action to high action against the same cell lines.

Chemical Composition, Antioxidant Capacity, and Anticancerous Effects against Human Lung Cancer Cells of a Terpenoid-Rich Fraction of Inula viscosa.

Inula viscosa is a widely used plant in traditional Mediterranean and Middle Eastern medicine for various illnesses. I. viscosa has been shown to have anticancer effects against various cancers, but its effects against lung cancer have been under limited investigation. At the same time, I. viscosa is rich in terpenoids whose anti-lung cancer effects have been poorly investigated. This study aimed to examine the potential anticancer properties of methanolic and aqueous extracts of stems and leaves of I. viscosa and its terpenoid-rich fraction against human lung cancer A549 cells. Results showed that the methanolic extracts of I. viscosa had significantly higher polyphenol and flavonoid content and radical scavenging capacity than the aqueous extracts. In addition, leaves methanolic extracts (IVLM) caused the highest reduction in viability of A549 cells among all the extracts. IVLM also reduced the viability of human ovarian SK-OV-3, breast MCF-7, liver HepG2, and colorectal HCT116 cancer cells. A terpenoid-rich I. viscosa fraction (IVL DCM), prepared by liquid-liquid separation of IVLM in dichloromethane (DCM), displayed a substantial reduction in the viability of A549 cells (IC50 = 27.8 ± 1.5 µg/mL at 48 h) and the panel of tested cancerous cell lines but was not cytotoxic to normal human embryonic fibroblasts (HDFn). The assessment of IVL DCM phytochemical constituents using GC-MS analysis revealed 21 metabolites, highlighting an enrichment in terpenoids, such as lupeol and its derivatives, caryophyllene oxide, betulin, and isopulegol, known to exhibit proapoptotic and antimetastatic functions. IVL DCM also showed robust antioxidant capacity and decent polyphenol and flavonoid contents. Furthermore, Western blotting analysis indicated that IVL DCM reduced proliferation (reduction of proliferation marker Ki67 and induction of proliferation inhibitor proteins P21 and P27), contaminant with P38 MAP kinase activation, and induced the intrinsic apoptotic pathway (P53/BCL2/BAX/Caspase3/PARP) in A549 cells. IVL DCM also reduced the migration of A549 cells, potentially by reducing FAK activation. Future identification of anticancer metabolites of IVL DCM, especially terpenoids, is recommended. These data place I. viscosa as a new resource of herbal anticancer agents.

Innovative Therapeutic Delivery of Metastasis-Associated in Colon Cancer 1-Suppressing miRNA Using High Transmembrane 4 L6 Family Member 5-Targeting Exosomes in Colorectal Cancer Mouse Models.

Elevated metastasis-associated in colon cancer 1 (MACC1) expression in colorectal cancer patients, and high transmembrane 4 L6 family member 5 (TM4SF5) protein expressed on various solid tumors' surface, are linked to aggressive cancer behavior and progression. In this study, adipose-derived stem cells (ASCs) were engineered to produce exosomes (Ex) that target the TM4SF5 protein on tumors. Moreover, MACC1-targeting microRNA was encapsulated within the Ex, resulting in TM4SF5-targeting Ex (MACC1-suppressing miRNA; miR-143). The anticancer effects of these Ex were investigated in vitro using the human colorectal cell line HCT116 and in vivo using colorectal cancer mouse xenograft models. In the in vivo assessment, administration of TM4SF5-targeting Ex[miR-143], referred to as tEx[miR-143] herein, resulted in the smallest tumor size, the lowest tumor growth rate, and the lightest excised tumors compared to other treatments (p < 0.05). It also led to the decreased expression of MACC-1 and anti-apoptotic markers MCL-1 and Bcl-xL while inducing the highest expression of pro-apoptotic markers BAX and BIM. These results were consistent with in vitro findings, where t Ex[miR-143] demonstrated the highest inhibition of HCT116 cell migration and invasion. These findings highlight the potential of tEx[miR-143] as an effective therapeutic strategy for colorectal cancer, demonstrating promising results in both targetability and anti-tumor effects in vitro and in vivo, warranting further investigation in clinical settings.

Synthesis of Cytotoxic Benzofurans and Ethers Derivatives of Paeonol.

Paeonol is a broadly studied natural product due to its many biological activities. Using a methodology previously employed by our research group, 11 derivatives of paeonol were synthesized (seven of them are unpublished compounds), including four ethers and seven benzofurans. Additionally, we determined the crystal structure of one of these ether derivatives (1 a) and of five benzofuran derivatives (2 a, 2 b, 2 c, 2 f and 2 g) by single crystal X-ray diffraction. To continue studying the cytotoxicity of this natural product and its derivatives, all compounds were tested against two cancer cell lines, HCT116 and MCF-7. Compounds 2 b, 2 e, and 2 g were considered active against the colorectal adenocarcinoma cells HCT116 (Growth inhibition >60 %). Compound 2 e showed an IC50 of 0.2 μM and was selected for further analysis, results reinforce its anticancer potential.

Synthesis and in vitro antitumor evaluation of new thieno[2,3-d]pyrimidine derivatives as EGFR and DHFR inhibitors

New thienopyrimidine derivatives 2–16 have been synthesized and their in vitro cytotoxicity was evaluated against five different human cancer cell lines HCT-116, Hela, MDA-MB-231, MCF7 and PC3. Compounds 6e, 7a, 7b, 7d, 10c and 10e displayed the highest antitumor activity against all tested cell lines compared to Doxorubicin. Enzyme inhibition assay revealed that compounds 6e and 10e showed high inhibitory activity against EGFR-TK, with IC50 values of 0.133 and 0.151 µM, compared to Olmutinib (IC50 = 0.028 µM); while the highest DHFR inhibitory activity was shown by compounds 7d and 10e with IC50 values of 0.462 and 0.541 µM, compared to Methotrexate (IC50 = 0.117 µM). Cell cycle analysis following a flow cytometric study using colorectal HCT-116 cancer cell line proved that compound 6e induced cell cycle arrest in G0-G1 phase, while compound 10e arrested the cell cycle at both G0-G1 and S phases. Additionally, both compounds (6e and 10e) were potently able to induce apoptosis in HCT-116 cell line. Docking results of compounds 6e and 10e into the pocket of EGFR active site showed their similar main binding features with Olmutinib, while compounds 7d and 10e showed only moderate fitting into DHFR compared to methotrexate. In silico studies revealed that most of the tested compounds obeyed Lipinski’s RO5 and showed positive drug likeness scores.

Unveiling the anticancer potential of the ethanolic extract from Trichoderma asperelloides.

The discovery of new therapeutic alternatives for cancer treatment is essential for improving efficacy and specificity, overcoming resistance, and enabling a more personalized approach for each patient. We investigated the antitumor activity of the crude ethanolic extract of the fungus Trichoderma asperelloides (ExtTa) and its interaction with chemotherapeutic drugs. It was observed, by MTT cytotoxicity assay, that ExtTa significantly reduced cell viability in breast adenocarcinoma, glioblastoma, lung carcinoma, melanoma, colorectal carcinoma, and sarcomas cell lines. The highest efficacy and selectivity of ExtTa were found against glioblastoma T98G and colorectal HCT116 cell lines. ExtTa is approximately four times more cytotoxic to those tumor cells than to non-cancer cell lines. A synergistic effect between ExtTa and doxorubicin was found in the treatment of osteosarcoma Saos-2 cells, as well as with 5-fluorouracil in the treatment of HCT116 colorectal carcinoma cells using CompuSyn software. Our data unravel the presence of bioactive compounds with cytotoxic effects against cancer cells present in T. asperelloides ethanolic crude extract, with the potential for developing novel anticancer agents.

The cytotoxicity of palladium (II) complexes containing 1,2‐ or 1,4‐diazine bridging ligands on squamous cell carcinoma cells in vitro: Insights in the mechanisms of action

Binuclear [{Pd (dien)}2(μ‐pz)](ClO4)4 (Pd1) and [{Pd (dien)}2(μ‐pydz)](ClO4)4 (Pd2) complexes, where pz is 1,4‐diazine and pydz is 1,2‐diazine bridging ligands and dien (diethylenetriamine) is a tridentately coordinated triamine ligand, were synthesized and characterized using elemental analysis, NMR (1H and 13C) spectroscopy, and UV–Vis spectrophotometry. Electronic absorption spectrophotometry and fluorescence spectroscopy were used to study the interactions of the prepared Pd1 and Pd2 complexes with deoxyribonucleic acid (CT‐DNA) and bovine serum albumin (BSA). According to the UV–Vis spectrophotometric data, the complexes interact with CT‐DNA and bind to the BSA as a possible carrier to the target site. The antitumor properties of the tested palladium compounds were evaluated on three types of human cancer cells, namely, squamous cell carcinoma cells SCC‐4, cervical carcinoma cells HeLa and colorectal carcinoma cells HCT116, and one type of malignantly unchanged cells, MRC‐5 fibroblasts. The highest cytotoxicity was shown by the Pd1 compound against SCC‐4 cell, with an IC50 value of 5.4 ± 1.2 μM. The results showed that the cytotoxic potential against SCC‐4 cells of our new palladium complexes Pd1 and Pd2 was comparable with the cytotoxic potential of cisplatin and oxaliplatin. However, Pd1 and Pd2 displayed significantly lower cytotoxicity in comparison with cisplatin and oxaliplatin against non‐malignant cells, fibroblasts MRC5, so these results may suggest that local and systemic complications of antitumor therapy could be reduced and/or avoided. Both compounds selectively decreased the viability of SCC‐4 cells by induction of apoptosis. Also, Pd1 and Pd2 induced S‐phase cell cycle arrest in human SCC‐4 cells, so that could be another mechanism to halt the uncontrolled growth and division of cancer cells. Moreover, the precise molecular mechanism of Pd1 and Pd2 complexes‐induced apoptosis was the activation of the intrinsic signaling pathway of apoptosis, which also included activation of caspase‐3. Therefore, our new palladium (II) complexes might have a promising role as the potential future cytostatic agents.

P53 Contributes to the Chemotherapeutic Drug Doxorubicin-Induced Cell Death in Colorectal Cancer Cell Line HCT116

Doxorubicin is a commonly used chemotherapy drug for cancer treatment, although its effectiveness varies across different cancer types. p53 is a key factor involved in cell death induced by therapeutic agents, and it can be upregulated by doxorubicin, exhibiting a function of apoptosis. To further investigate the mechanism between p53 and doxorubicin, this study explored whether p53 plays a role in doxorubicin-induced cell death in the colorectal cancer line HCT116. The findings revealed that p53 was upregulated in HCT116 cells when treated with doxorubicin, and the knockdown of p53 decreased the sensitivity of HCT116 cells to doxorubicin. These results suggest that p53 plays an important role in doxorubicin-induced cell death in HCT116 cells, potentially contributing to more effective treatment approaches.

Pt(iv) derivatives of cisplatin and oxaliplatin bearing an EMT-related TMEM16A/COX-2-selective dual inhibitor against colorectal cancer cells HCT116.

Colorectal cancer represents the over-expression of TMEM16A and COX-2, offering a promising therapeutic strategy. Two Pt(iv) conjugates derived from Pt(ii) drug (cisplatin or oxaliplatin) and niflumic acid, complexes 1 and 2, were designed and prepared to exert the positive impact of multiple biological targets of DNA/TMEM16A/COX-2 against colorectal cancer. Complex 2 afforded higher cytotoxicity than 1 and the combination of an intermediate of oxidized oxaliplatin and NFA against cancer cells A549, HeLa, MCF-7, and HCT116. Especially for colorectal cancer cells HCT116, 2 was significantly more toxic (22-fold) and selective to cancer cells against normal HUVEC cells (4-fold) than first-line oxaliplatin. The outstanding anticancer activity of 2 is partly attributed to its dramatic increase in cellular uptake, DNA damage, and apoptosis. Mechanistic studies indicated that 2 inhibited HCT116 cell metastasis by triggering TMEM16A, COX-2, and their downstream signaling pathways, including EGFR, STAT3, E-cadherin and N-cadherin.