Cell Line COLO Research Articles

Assessment of metabolic stability, pharmacokinetics by LC-MS/MS and establishment of safe dose of IMID-2, a novel anticancer molecule under drug discovery.

Pyruvate kinase (PK) M2 activators ramp up glycolysis in cancer cells leading to a reversal of the Warburg effect in cancer cells. A promising PKM2 activator molecule IMID-2 developed by National Institute of Pharmaceutical Education & Research, Ahmedabad (NIPER-A) showed promising anticancer activity against MCF-7 and COLO-205 cell lines which represent breast and colon cancer. Its physicochemical properties like solubility, ionization constant, partition coefficient, and distribution constant have already been established. Its metabolic pathway is also well established through in vitro and in vivo metabolite profiling and reported previously. In this study, we have evaluated the metabolic stability of IMID-2 using (LC-MS/MS and investigated the safety aspect of the molecule through an acute oral toxicity study. In vivo studies in rats confirmed that the molecule is safe even at the dose level of 175 mg/kg. Furthermore, the pharmacokinetic study of IMID-2 was also carried out by LC-MS/MS to understand its ADME profile. The molecule was found to have promising bioavailability through the oral route. This research work is thus another step toward drug-likeness testing of this promising anti-cancer molecule. The molecule can be considered to be a potential anti-cancer lead based on the earlier report substantiated by current findings.

Design, Synthesis, and Anticancer Evaluation of Novel Tetracaine Hydrazide-Hydrazones.

Tetracaine is an ester derivative used as a local anesthetic molecule. In this study, a series of novel Tetracaine derivatives bearing hydrazide-hydrazone moiety were designed, synthesized, and evaluated for anticancer activity. The structures of these compounds were characterized by spectral (1H NMR,13C NMR, FT-IR, and HRMS analyses) methods. All synthesized compounds were screened for anticancer activity against two different human cancer cell lines (Colo-205 and HepG2). Among the synthesized molecules, compounds 2f and 2m showed the most potent anticancer activity against the Colo-205 cell line (IC50 = 50.0 and 20.5 μM, respectively). Compounds 2k, 2p, and 2s demonstrated the best anticancer activity against the HepG2 cell line (IC50 = 30.5, 35.9, and 20.8 μM, respectively). mRNA transcription levels of Bax and caspase-3 genes were determined by real-time polymerase chain reaction (qRT-PCR) analysis of both Colo-205 and HepG2 cell lines. Doxorubicin was used as a positive sensitivity reference standard. qRT-PCR analysis showed that there was a time-dependent rise in the expression levels of Bax and Caspase 3 on apoptosis. Inhibition of apoptotic proteins PI3K, Akt, PTEN, pPTEN, FoXO1, FoXO3a, TXNIP, and p27 was investigated in Colo-205 and HepG2 cells treated with compounds 2f, 2m, 2k, 2p, and 2s by using Western blotting.

Synthesis and biological evaluation of benzamide-chalcone hybrids as potential c-Metkinase and COX-2 inhibitors.

c-Met kinase and cyclooxygenase 2 (COX-2) enzymes are two significant targets in tumor progression. Chalcone and benzamide moieties were combined using molecular hybridization to assess their potential as c-Met kinase and COX-2 inhibitors. 4-Methylbenzamide and 4-chlorobenzamide chalcone analogs were synthesized, characterized, and evaluated for antiproliferative activity on Michigan Cancer Foundation-7 (MCF-7), HT-29, MDA-MB-231, COLO-205, and A549 cell lines by sulforhodamine-B stain (SRB) assay. Following the SRB assay, compounds were evaluated for their c-Met kinase and COX-2 inhibitory potential. All compounds inhibited COX-2 with half-maximal inhibitory concentration (IC50 ) <10 µM. Compounds 7h, 7i, 7j, 8f, and 8j inhibited c-Met with IC50 <10 µM. Compound 7h was evaluated for its long-term antiproliferative and anti-migratory effects by colony formation and wound healing assay. It exerted these effects in a concentration-dependent manner. Compounds 7j and 8j were further evaluated for in vitro antiangiogenic effects. Compound 7j exhibited moderate antiangiogenic effect while compound 8j exhibited strong effect. Compounds 7h, 7i, 7j, 8f, and 8j were evaluated for the serum protein binding, using the in vitro bovine serum albumin binding assay. The results indicated that the tested compounds bind to bovine serum albumin (BSA) and can be further explored by other studies.

Half-Sandwich Rhodium Complexes with Releasable N-Donor Monodentate Ligands: Solution Chemical Properties and the Possibility for Acidosis Activation.

Cancer chemotherapeutics usually have serious side effects. Targeting the special properties of cancer and activation of the anticancer drug in the tumor microenvironment in situ may decrease the intensity of the side effects and improve the efficacy of therapy. In this study, half-sandwich Rh complexes are introduced, which may be activated at the acidic, extracellular pH of the tumor tissue. The synthesis and aqueous stability of mixed-ligand complexes with a general formula of [Rh(η5-Cp*)(N,N/O)(N)]2+/+ are reported, where (N,N/O) indicates bidentate 8-quinolate, ethylenediamine and 1,10-phenanthroline and (N) represents the releasable monodentate ligand with a nitrogen donor atom. UV-visible spectrophotometry, 1H NMR, and pH-potentiometry were used to determine the protonation constants of the monodentate ligands, the proton dissociation constants of the coordinated water molecules in the aqua complexes, and the formation constants of the mixed-ligand complexes. The obtained data were compared to those of the analogous Ru(η6-p-cymene) complexes. The developed mixed-ligand complexes were tested in drug-sensitive and resistant colon cancer cell lines (Colo205 and Colo320, respectively) and in four bacterial strains (Gram-positive and Gram-negative, drug-sensitive, and resistant) at different pH values (5-8). The mixed-ligand complexes with 1-methylimidazole displayed sufficient stability at pH 7.4, and their activation was found in cancer cells with decreasing pH; moreover, the mixed-ligand complexes demonstrated antimicrobial activity in Gram-positive and Gram-negative bacteria, including the resistant MRSA strain. This study proved the viability of incorporating releasable monodentate ligands into mixed-ligand half-sandwich complexes, which is supported by the biological assays.

Properties of Linum usitatissimum Seed Aqueous Extract Against Colon Cancer Cell Lines

Background: Colon cancer has been a rising health concern in the modern world for quite some time. Considering the harmful side effects of chemotherapy, the most common treatment for this cancer, new treatment methods are required to lessen the side effects of treatment on patients. Traditional and herbal medicine can help in this effort. Objectives: Our study investigated the antiproliferative and pro-apoptotic effects of Linum usitatissimum seed aqueous extract on colon cancer and the expression of apoptotic genes compared to normal cell lines to reduce chemotherapy's harmful and irreversible side effects on colon cancer patients. Methods: MTT assay was used to determine the cytotoxicity of our extract on LS174T and COLO205 cells. Cancer and normal cell lines were treated with 2, 4, 6, and 8 mg/mL of LU extract for 24 and 48 hours. RNA extraction was performed, and mRNAs were reverse transcribed to cDNAs using RT-PCR. qPCR was then performed to assess the expression of BAX and BAD genes. Results: MTT assay concluded that this extract has a good cytotoxic effect on LS174t and COLO205 cell lines and can decrease cell viability, while no significant decrease in cell viability was observed in normal cells. For qPCR assay, the cells were treated with 0.5, 1, and 2 mg/mL of LU extract for two days. The qPCR analysis revealed that the expression of pro-apoptotic BAX and BAD was significantly decreased following treatment with our extract, while the expression of these genes showed no significant changes in normal cell lines after the same treatment. Conclusions: These results show that the components within L. usitatissimum seed aqueous extract have antiproliferative and pro-apoptotic properties. This extract can successfully decrease the population of LS174t and COLO205 cells while increasing the expression of pro-apoptotic BAX and BAD in these cell lines. The results also demonstrate that normal non-cancerous cells are unharmed by this extract. These results hint at some potential anti-cancer properties of L. usitatissimum seed aqueous extract.

In Vitro and In Silico Biological Studies of 4-Phenyl-2-quinolone (4-PQ) Derivatives as Anticancer Agents

Our previous study found that 2-phenyl-4-quinolone (2-PQ) derivatives are antimitotic agents, and we adopted the drug design concept of scaffold hopping to replace the 2-aromatic ring of 2-PQs with a 4-aromatic ring, representing 4-phenyl-2-quinolones (4-PQs). The 4-PQ compounds, whose structural backbones also mimic analogs of podophyllotoxin (PPT), maybe a new class of anticancer drugs with simplified PPT structures. In addition, 4-PQs are a new generation of anticancer lead compounds as apoptosis stimulators. On the other hand, previous studies showed that 4-arylcoumarin derivatives with 5-, 6-, and 7-methoxy substitutions displayed remarkable anticancer activities. Therefore, we further synthesized a series of 5-, 6-, and 7-methoxy-substituted 4-PQ derivatives (19-32) by Knorr quinoline cyclization, and examined their anticancer effectiveness. Among these 4-PQs, compound 22 demonstrated excellent antiproliferative activities against the COLO205 cell line (50% inhibitory concentration (IC50) = 0.32 μM) and H460 cell line (IC50 = 0.89 μM). Furthermore, we utilized molecular docking studies to explain the possible anticancer mechanisms of these 4-PQs by the docking mode in the colchicine-binding pocket of the tubulin receptor. Consequently, we selected the candidate compounds 19, 20, 21, 22, 25, 27, and 28 to predict their absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiles. Pharmacokinetics (PKs) indicated that these 4-PQs displayed good drug-likeness and bioavailability, and had no cardiotoxic side effects or carcinogenicity, but we detected risks of drug-drug interactions and AMES toxicity (mutagenic). However, structural modifications of these 4-PQs could improve their PK properties and reduce their side effects, and their promising anticancer activities attracted our attention for further studies.

Synthesis and biological evaluation of coumarin-quinone hybrids as multifunctional bioactive agents.

We report the synthesis, structural characterization and pharmaceutical activity of four coumarin-quinone hybrids. The compounds were significantly active against Staphylococcus aureus, Pseudomonas aeoginosa and Candida albicans. Promising antioxidant activity was observed when compared to ascorbic acid. Two compounds, DTBSB and DTBSN, also showed commendable in vitro antiproliferative activities against the cells of human cancer cell lines MCF-7, MDA-MB-231, COLO-205, HT-29 and A549 along with appreciable tumor selectivity with distinct selectivity index. Molecular docking studies using cyclooxygenase-2 (PDB ID: 6COX) revealed strong binding affinities for the COX-2 active site. Moreover, ADMET properties of the synthesized compounds were determined using the pKCSM and SwissADME online tools and all the compounds had accurate pharmacokinetic profiles. Hence, the new coumarin-quinone hybrids DTBSB and DTBSN can be considered for optimization and lead development.

Indolo[2,3-e]benzazocines and indolo[2,3-f]benzazonines and their copper(II) complexes as microtubule destabilizing agents.

A series of four indolo[2,3-e]benzazocines HL1-HL4 and two indolo[2,3-f]benzazonines HL5 and HL6, as well as their respective copper(II) complexes 1-6, were synthesized and characterized by 1H and 13C NMR spectroscopy, ESI mass spectrometry, single crystal X-ray diffraction (SC-XRD) and combustion analysis (C, H, N). SC-XRD studies of precursors Vd, VIa·0.5MeOH, of ligands HL4 and HL6·DCM, and complexes 2·2DMF, 5·2DMF, 5'·iPrOH·MeOH provided insights into the energetically favored conformations of eight- and nine-membered heterocycles in the four-ring systems. In addition, proton dissociation constants (pKa) of HL1, HL2 and HL5, complexes 1, 2 and 5, overall stability constants (log β) of 1, 2 and 5 in 30% (v/v) DMSO/H2O at 298 K, as well as thermodynamic solubility of HL1-HL6 and 1-6 in aqueous solution at pH 7.4 were determined by UV-vis spectroscopy. All compounds were tested for antiproliferative activity against Colo320, Colo205 and MCF-7 cell lines and showed IC50 values in the low micromolar to sub-micromolar concentration range, while some of them (HL1, HL5 and HL6, 1, 2 and 6) showed remarkable selectivity towards malignant cell lines. Ethidium bromide displacement studies provided evidence that DNA is not the primary target for these drugs. Rather, inhibition of tubulin assembly is likely the underlying mechanism responsible for their antiproliferative activity. Tubulin disassembly experiments showed that HL1 and 1 are effective microtubule destabilizing agents binding to the colchicine site. This was also confirmed by molecular modelling investigations. To the best of our knowledge, complex 1 is the first reported transition metal complex to effectively bind to the tubulin-colchicine pocket.

Screening of 3-acetylcoumarin derivatives as multifunctional biological agents

In this work, the multifunctional potential of four 3-acetylcoumarin derivatives was studied. The derivatives were significantly active against bacteria Staphylococcus aureus, Pseudomonas aeoginosa and fungal strain Candida albicans. The results of antioxidant activity assays were promising when compared to ascorbic acid. The in vitro anticancer activity was carried out using MTT assay on human cancer cell line COLO-205 and 3ACDT showed commendable antiproliferative activity along with appreciable tumor selectivity with distinct selectivity index. Moreover, ADMET properties of the compounds were determined using the pKCSM and SwissADME online tools and all compounds were found with good pharmacokinetic profile. Hence, from the obtained results from all the 3-acetylcoumarin derivatives, 3ACDT exhibited good therapeutical potential and can be optimized for lead development.

Coumarins, furocoumarins and limonoids of Citrus trifoliata and their effects on human colon adenocarcinoma cell lines

Citrus trifoliata L. (Chinese or Japanese bitter orange) is a medicinal plant with furocoumarins and limonoids as characteristic secondary metabolites. The bitter taste of the fruit limits its use as food, however, it is applied in Asian traditional medicine for its antiphlogistic effect, to treat digestive ulcers and different gastrointestinal disorders and cancer. The phytochemical composition and pharmacological characteristics of this species have not been fully discovered, nevertheless its potential antiproliferative or cytotoxic effects might be related to furocoumarins or limonoids.Our aim was to isolate and identify secondary metabolites from C. trifoliata peel and seeds and to investigate their bioactivities that might be related to the supposed anticancer effect of the plant. By using different chromatographic methods, six pure compounds (phellopterin (2), scoparone (3), myrsellin (4), triphasiol (6), umbelliferone (7) and citropten (5,7-dimethoxycoumarin (8)) were isolated from the peel and four (imperatorin (1), auraptene (5), limonin (9) and deacetyl nomilin (10)) from the seeds of C. trifoliata fruits. These compounds are furocoumarin (1, 2), coumarin (3–8), and limonoid derivatives (9, 10). Scoparone (3) has been detected in this species for the first time.The furocoumarins (1–2) showed moderate activity on the human colorectal adenocarcinona tumor cell line COLO 320 in antiproliferative assays and 2 also had remarkable P-glycoprotein inhibitory activity and synergistic effect with doxorubicin. The coumarin 5 showed significant activity on the COLO 320 cell line in antiproliferative assays and P-glycoprotein inhibitory activity in the FACS (fluorescence activated cell sorting) assay.

Antiproliferative and cytotoxic effects of sesquiterpene lactones isolated from Ambrosia artemisiifolia on human adenocarcinoma and normal cell lines

Context Ambrosia artemisiifolia L. (Asteraceae) contains sesquiterpene lactones as characteristic secondary metabolites. Many of these compounds exert antiproliferative and cytotoxic effects. Objective To isolate the sesquiterpene lactones from the aerial part of A. artemisiifolia and to elucidate their cytotoxic, antiproliferative and antibacterial effects. Materials and methods The compounds were identified by one-dimensional (1D) and 2D NMR, HR-MS spectroscopy from the methanol extract. Isolated compounds were investigated for their cytotoxic and antiproliferative effects on human colonic adenocarcinoma cell lines and human embryonal lung fibroblast cell line using MTT assay. The selectivity of the sesquiterpenes was calculated towards the normal cell line. To check the effect of drug interactions between compounds and doxorubicin, multidrug-resistant Colo 320 cells were used. Results A new seco-psilostachyinolide derivative, 1,10-dihydro-1′-noraltamisin, and seven known compounds were isolated from the methanol extract. Acetoxydihydrodamsin had the most potent cytotoxic effect on sensitive (Colo205) cell line (IC50 = 7.64 µM), also the strongest antiproliferative effect on Colo205 (IC50 = 5.14 µM) and Colo320 (IC50 = 3.67 µM) cell lines. 1′-Noraltamisin (IC50 = 8.78 µM) and psilostachyin (IC50 = 5.29 µM) showed significant antiproliferative effects on the multidrug-resistant Colo320 cell line and had moderate selectivity against human embryonal lung fibroblast cell line. Psilostachyin C exhibited cytotoxic effects on Colo205 cells (IC50 = 26.60 µM). None of the isolated compounds inhibited ABCB1 efflux pump (EP; P-glycoprotein) or the bacterial EPs. Discussion and conclusions Acetoxydihydrodamsin, 1′-noraltamisin, and psilostachyin showed the most remarkable cytotoxic and antiproliferative activity on tumour cell lines and exerted selectivity towards MRC-5 cell line.

Heliotropium ramosissimum metabolic profiling, in silico and in vitro evaluation with potent selective cytotoxicity against colorectal carcinoma

Heliotropium is a genus of the Boraginaceae family. Its members are used in many traditional and folklore medicines to treat several ailments. Despite this widespread usage, only a few evidence-based scientific studies investigated and identified its phytoconstituents. Herein, we documented the chemical profile of the Heliotropium ramosissimum methanolic extract using gas chromatography-mass spectrometry (GC–MS) and liquid chromatography-tandem mass spectrometry (LC–ESI–MS/MS) and assessed its antioxidant and cytotoxic effects. The methanolic extract exhibited high phenolic content (179.74 ± 0.58 µg/mL) and high flavonoid content (53.18 ± 0.60 µg/mL). The GC–MS analysis of the lipoidal matter allowed us to identify 41 compounds with high percentages of 1,2-benzenedicarboxylic acid, bis(2-methoxyethyl) ester (23.91%), and 6,10,14-trimethylpentadecan-2-one (18.74%). Thirty-two phytomolecules were tentatively identified from the methanolic extract of H. ramosissimum using LC–MS/MS. These compounds belonged to several phytochemical classes such as phenolic acids, alkaloids, coumarins, and flavonoids. Furthermore, we assessed the antioxidant activity of the methanolic extract by DPPH assay and oxygen radical absorbance capacity assay, which yielded IC50 values of 414.30 µg/mL and 170.03 ± 44.40 µM TE/equivalent, respectively. We also assessed the cytotoxicity of the methanolic extract on seven different cell lines; Colo-205, A-375, HeLa, HepG-2, H-460, and OEC showed that it selectively killed cancer cells with particularly potent cytotoxicity against Colo-205 without affecting normal cells. Further studies revealed that the extract induced apoptosis and/or necrosis on Colo-205 cell line at an IC50 of 18.60 µg/mL. Finally, we conducted molecular docking on the LC–ESI–MS/MS-identified compounds against colon cancer antigen 10 to find potentially cytotoxic compounds. Binding score energy analysis showed that isochlorogenic acid and orientin had the highest affinity for the colon cancer antigen 10 protein, with binding scores of (− 13.2001) and (− 13.5655) kcal/mol, respectively. These findings suggest that Heliotropium ramosissimum contains potent therapeutic candidates for colorectal cancer treatment.

Anti-proliferative Effects of Ocimum basilicum Leaf Aqueous Extract on Colon Cancer Cell Lines and the Expression of Apoptotic Genes

Background: Colon cancer is one of the most diagnosed cancers in the modern world. Chemotherapy is one of the most relevant treatments for colon cancer, but this method of treatment has severe and sometimes irreversible side effects. Therefore, alternative treatments with fewer side effects are highly considered. Natural and herbal medicine has been proven to be a great source for these alternative treatment methods. Objectives: This study aimed to investigate the anti-proliferative and cytotoxic effects of Ocimum basillicum (OB) leaf aqueous extract on human colon cancer cell lines LS174T and COLO205 and the expression of apoptotic genes. Methods: Methyl thiazol tetrazolium (MTT) assay was performed to assess the cytotoxic effects of our extract on the aforementioned cell lines. The changes in the expression of BAX and BAD were analyzed by qPCR following the treatment of cells with our extract. Results: The results of the MTT assay indicated that the extract had a cytotoxic effect and could reduce the population of live cancer cells, especially LS174T cells. The expression of BAX and BAD genes showed a significant increase in qPCR analysis in both cell lines, especially in the LS174T cells. Conclusions: Our results showed that the extract had cytotoxic, anti-proliferative, and pro-apoptotic characteristics on previously mentioned cell lines, especially on the LS 174T cells.

IN VITRO ANTICANCER ACTIVITY OF LEAF EXTRACT OF COUROUPITA GUIANENSIS AGAINST COLON CANCER CELL LINE

Objective: This study aims in dealing with the in vitro anticancer activity of leaves of Couroupita guianensis (family: Lecythidaceae) against Colon Cancer cell line(Colo205). Materials and Methods: The in vitro anticancer activity of methanolic leaf extract of Couroupita guianensis was studied against Colon cancer cell line(Colo205) using MTT Assay. Results: The methanolic extract of leaves (CgL)of Couroupita guianensis showed good percentage of inhibition against Colo205 cell line as compared to standard drug. Conclusion: The study provide information which may generate interest among researchers.

Synthesis of novel isoxazole–carboxamide derivatives as promising agents for melanoma and targeted nano-emulgel conjugate for improved cellular permeability

BackgroundCancer is one of the most dangerous and widespread diseases in the world today and it has risen to the position of the leading cause of death around the globe in the last few decades. Due to the inherent resistance of many types of cancer to conventional radiotherapy and chemotherapy, it is vital to develop innovative anticancer medications. Recently, a strategy based on nanotechnology has been used to improve the effectiveness of both old and new cancer drugs.ObjectivesThe present study aimed to design and synthesize a series of phenyl-isoxazole–Carboxamide derivatives, evaluate their anticancer properties, and improve the permeability of potent compounds into cancer cells by using a nano-emulgel strategy.MethodsThe coupling reaction of aniline derivatives and isoxazole–Carboxylic acid was used to synthesize a series of isoxazole–Carboxamide derivatives. IR, HRMS, 1H-NMR, and 13C-NMR spectroscopy techniques, characterized all the synthesized compounds. The in-vitro cytotoxic evaluation was performed by using the MTS assay against seven cancer cell lines, including hepatocellular carcinoma (Hep3B and HepG2), cervical adenocarcinoma (HeLa), breast carcinoma (MCF-7), melanoma (B16F1), colorectal adenocarcinoma (Caco-2), and colon adenocarcinoma (Colo205), as well as human hepatic stellate (LX-2) in addition to the normal cell line (Hek293T). A nano-emulgel was developed for the most potent compound, using a self-emulsifying technique.ResultsAll synthesized compounds were found to have potent to moderate activities against B16F1, Colo205, and HepG2 cancer cell lines. The results revealed that the 2a compound has broad spectrum activity against B16F1, Colo205, HepG2, and HeLa cancer cell lines with an IC50 range of 7.55–40.85 µM. Moreover, compound 2e was the most active compound against B16F1 with an IC50 of 0.079 µM compared with Dox (IC50 = 0.056 µM). Nanoemulgel was used to increase the potency of the 2e molecule against this cancer cell line, and the IC50 was reduced to 0.039 µM. The antifibrotic activities were investigated against the LX-2 cell line, and it was found that our synthesized molecules showed better antifibrotic activities at 1 µM than 5-FU, and the cell viability values were 67 and 95%, respectively.ConclusionThis study suggests that a 2e nano-formalized compound is a potential and promising anti-melanoma agent.

Synthesis, characterization and molecular docking studies of phenoxyimine based ligands: Cytotoxicity, hemolytic activity and antioxidant assessment

A series of phenoxyimine compounds were designed and synthesized by the condensation of different substituted salicylaldehyde and aniline at the molar ratio of 1:1 in one step reaction. All synthesized compounds were characterized by analytical and spectroscopic techniques. All compounds were screened for cytotoxicity activities against different cancer cell lines. Compound 3 exhibits significant cytotoxic activity with an IC50 value of 1.099 µM, 1.132 µM against COLO-205 (Colorectal adenocarcinoma) and A-549 (Lung carcinoma) cell line respectively, while compound 4 exhibits noteworthy cytotoxic activity with an IC50 value of 0.52 µM against MDA-MB-231(Breast adenocarcinoma). Similarly, other compounds also exhibited good cytotoxicity properties. Furthermore, the phenoxyimine compounds showed the greatest antioxidant activity against DPPH radicals. The nontoxic activity was confirmed by cytocompatibility assay on L929 normal cell line and hemolysis assay on human red blood cells. Additionally, in silico molecular docking of all compounds were carried out against EGFR, HER-2 and VEGF. Compound 3 having minimum binding energy ΔGb = -8.81 kcal/mol, ΔGb = -10.49 kcal /mol, ΔGb = -9.99 kcal /mol, was bound into the active pocket of EGFR, HER-2 and VEGF respectively.

P-305 Oxidized-LDL promotes colorectal cancer progression and growth of human colonoides

Expression levels of the oxidized-LDL receptor LOX-1 have been related to cancer progression in prostatic, gastric, pancreatic, and colorectal cancer (CRC); however, the relationship among pro-inflammatory markers, ox-LDL/LOX-1 and CRC are not completely elucidated. We tested the hypothesis that stimulation of CRC cell lines with TNF-α and ox-LDL promotes tumor progression. We also explore the effect of ox-LDL on human colonoids growth. The colorectal cancer (CRC) cell lines HCT116 and COLO320 were treated with TNF-α and ox-LDL and tested for LOX-1 expression. The effect on cell proliferation of this proinflammatory signals were assayed by MTT and cell count with the exclusion dye trypan blue. LOX-1-negative cell lines were obtained by transduction with lentiviral vectors encoding a shRNA specific for LOX-1. The expression of genes involved in tumor progression were analyzed by qRT-PCR and Western blot in wild type and LOX-1 negative cells treated with TNF-α and ox-LDL. Cell migration and invasion was assayed in ox-LDL and TNF-α treated cells. Human colonoids were stimulated with ox-LDL in the presence of the NADPH oxidase inhibitors ML171, VAS2870 and ROS scavenger NAC. Surface area and survival were assessed by contrast microscopy and MTT. Proliferation was assessed by click-chemistry. LOX-1 receptor is expressed in the CRC cell lines HCT116 and COLO320. Additionally, we found in microarrays data stored in public databases that LOX-1 mRNA is over-expressed in colorectal carcinomas and adenocarcinomas compared with normal colon tissue. Treatment of CRC cell lines with TNF-α and ox-LDL augmented LOX-1 expression and LOX-1 silencing abrogated this effect. Moreover, both cell lines treated with ox-LDL proliferated in a dose-dependent manner. Importantly, ox-LDL stimulated human organoids proliferation and viability also in a dose-dependent manner, an effect partly mediated by ROS. The activation of LOX-1 by ox-LDL induced the expression of angiogenic markers such as MMP2, MMP7, MMP9 and VEGF. LOX-1 activation by oxLDL also promotes epithelial to mesenchymal transition of human colorectal cancer cells, characterized by reduced expression of the epithelial marker E-cadherin and increased expression of mesenchymal markers such as Vimentin, N-Cadherin, TWIST, ZEB1 and ZEB2. In addition, we determined that ox-LDL promotes invasion and migration of colorectal cancer cells. Nevertheless, co-treatment of colorectal cancer cells with TNF-α and ox-LDL did not show an additive effect. LOX-1 activation by ox-LDL induces a transcription reprogramming of genes involved in angiogenesis, invasion, migration, and cell proliferation. In the context of inflammation and obesity, we speculate that overexpression of LOX-1 provides additional stimulation accelerating tumor progression.

Heat stress stimulates colon cancer cells to secret specific population of extracellular nanovesicles enriched by HSP70 and microRNAs

Background. Heat stress (HS) induces the cellular secretion of heat shock proteins (HSP ) and extracellular nanovesicles (ENVs). The biological link between these phenomena is poorly understood. In the case of colorectal cancer (CRC) cells, the secretion of HSP s and ENV may be involved in the clinical response to intraperitoneal therapy of peritoneal carcinomatosis.Material and Methods. Established colon cancer cell lines COLO 320, HCT 116, HT29 and DLD 1 were used. ENVs were isolated from culture media by differential ultra-centrifugation and analyzed by dynamic light scattering, nanoparticle tracking analysis, atomic force microscopy and flow cytometry. Super-paramagnetic particles (SPMP ) covered by antibodies to the membrane form of Hsp70 were used for isolation and quantification of Hsp70(+) ENVs. Vesicular microRNA was assayed by RT-qPC R.Results. HS induces the secretion of ENVs by CRC cells, the resistance to HS correlates with the activity of HS-induced ENVs secretion. HS induces the secretion of a specific population of ENVs enriched by membrane form Hsp70 (mHsp70). The microRNA content of mHsp70(+) ENVs has qualitative and quantitative features. The concentration of miR-126-3p, -181-5p, -155-5p, -223 is increased in mHSP 70(+) ENVs secreted by three CRC cell lines.Conclusion. HS induces the secretion of mHSP 70(+) ENVs by CRC cells. This phenomenon may be involved in a clinical response to intraperitoneal chemo-hyperthermic perfusion therapy of peritoneal carcinomatosis.

Design, synthesis, and biological evaluation of chalcone based derivatives of 1,3,4-thiadazole-oxazole as cytotoxic agents

A new series of chalcone incorporated 1,3,4-thiadazole-oxazole (10a-j) compounds have been prepared and its chemical structures were confirmed by spectroscopic techniques. Further, all compounds were explored for cytotoxic properties alongside human cervix cancer (SiHa), human lung cancer (A549), human breast cancer (MCF-7) and human colon cancer (Colo-205) cell lines by employing MTT assay. The known chemotherapeutic drug candidate as etoposide used as positive control. According to the data, all the compounds showed good to moderate activities. Among them, compounds 10a, 10b, 10c and 10d displayed most promising activity.

Highly Antiproliferative Latonduine and Indolo[2,3-c]quinoline Derivatives: Complex Formation with Copper(II) Markedly Changes the Kinase Inhibitory Profile.

A series of latonduine and indoloquinoline derivatives HL1–HL8 and their copper(II) complexes (1–8) were synthesized and comprehensively characterized. The structures of five compounds (HL6, [CuCl(L1)(DMF)]·DMF, [CuCl(L2)(CH3OH)], [CuCl(L3)]·0.5H2O, and [CuCl2(H2L5)]Cl·2DMF) were elucidated by single crystal X-ray diffraction. The copper(II) complexes revealed low micro- to sub-micromolar IC50 values with promising selectivity toward human colon adenocarcinoma multidrug-resistant Colo320 cancer cells as compared to the doxorubicin-sensitive Colo205 cell line. The lead compounds HL4 and 4 as well as HL8 and 8 induced apoptosis efficiently in Colo320 cells. In addition, the copper(II) complexes had higher affinity to DNA than their metal-free ligands. HL8 showed selective inhibition for the PIM-1 enzyme, while 8 revealed strong inhibition of five other enzymes, i.e., SGK-1, PKA, CaMK-1, GSK3β, and MSK1, from a panel of 50 kinases. Furthermore, molecular modeling of the ligands and complexes showed a good fit to the binding pockets of these targets.