Human Cancer Cell Line HCT116 Research Articles

Abstract 186: Advances in 3D cell culture analysis: Utilizing novel flow cytometry technology for rapid analysis of intact spheroids

Abstract Cancer cells grown in two dimensional (2D) monolayers are a convenient model to use for basic research and in generating lead candidates in the drug discovery process. However, cells grown in 2D cultures lack many of the structural, biological, and functional attributes of solid tumors, which contributes to the high clinical failure rates for biologic and small molecule drugs. Multicellular three-dimensional (3D) tumor spheroids are advanced models which more faithfully mimic the physiological tumor milieu and is more predictive of therapeutic efficacy than traditional monolayer cultures. However, the inability to generate and to analyze large numbers of intact spheroids with a simplified workflow remains a barrier in the adoption of 3D models for a wide range of applications. To overcome these barriers, we used the Velocyt® large particle flow cytometer (Bennubio) to evaluate spheroid health in a well-established small molecule spheroid killing assay. The Velocyt technology rapidly and gently analyzes particles from 3 to 300 microns and comes with a suite of novel informatics tools purpose built for large particle analysis. To exemplify the use of large particle flow cytometry for the analysis of intact spheroids we generated uniformed-sized spheroids (~180 microns) from the H1650 human lung and HCT116 human colon cancer cell lines using microwell technology. Two to three days after spheroid generation, the spheroids were treated with increasing concentrations of staurosporine and other compounds. Spheroids were harvested at one to three days post treatment (depending on drug treatment) and stained with fluorescent cell viability and apoptotic dyes. Intact spheroids were analyzed for fluorescence and size using the Velocyt flow cytometer and between one and two thousand spheroids were assessed for each treatment in approximately three minutes. In untreated control spheroids, fluorescence was detected in a subpopulation of spheroids consistent with expected low levels of background apoptosis. In treated spheroids, an increase in the mean fluorescence intensity (MFI) of each particle indicated an increase in the number of non-viable and apoptotic cells in each spheroid. MFI increased nearly 6-fold with high concentrations of staurosporine compared to control spheroids, and dead cells were detected in nearly 100% of spheroids that were administered the highest staurosporine concentration. Using the size-based measurement parameters of the Velocyt, we also showed a decrease in overall spheroid size concurrent with increased numbers of spheroid fragments and single cells with increasing drug concentration. These data highlight that the use of 3D cell models combined with the Velocyt large particle flow cytometer is a powerful tool to help advance basic and translational cancer research. Citation Format: John P. O'Rourke, Mathew Saunders, Valerie Sanchez. Advances in 3D cell culture analysis: Utilizing novel flow cytometry technology for rapid analysis of intact spheroids [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 186.

New Terpenoids from Potentilla freyniana Bornm. and Their Cytotoxic Activities.

Two new A-ring contracted triterpenoids, madengaisu A and madengaisu B, and one undescribed ent-kaurane diterpenoid, madengaisu C, along with 20 known compounds were isolated from the roots of Potentilla freyniana Bornm. The structures were elucidated using extensive spectroscopic techniques, including 1D and 2D-NMR, HR-ESI-MS, ECD spectra, IR, and UV analysis. Moreover, all isolated constituents were evaluated for their anti-proliferative activity against RA-FLS cells and cytotoxic activities against the human cancer cell lines Hep-G2, HCT-116, BGC-823, and MCF-7. Ursolic acid and pomolic acid displayed moderate inhibitory activity in RA-FLS cells with IC50 values of 24.63 ± 1.96 and 25.12 ± 1.97 μM, respectively. Hyptadienic acid and 2α,3β-dihydroxyolean-12-en-28-oic acid 28-O-β-d-glucopyranoside exhibited good cytotoxicity against Hep-G2 cells with IC50 values of 25.16 ± 2.55 and 17.66 ± 1.82 μM, respectively. In addition, 2α,3β-dihydroxyolean-13(18)-en-28-oic acid and alphitolic acid were observed to inhibit HCT-116 cells (13.25 ± 1.65 and 21.62 ± 0.33 μM, respectively), while madengaisu B and 2α,3β-dihydroxyolean-13(18)-en-28-oic acid showed cytotoxic activities against BGC-823 cells with IC50 values of 24.76 ± 0.94 and 26.83 ± 2.52 μM, respectively, which demonstrated that triterpenes from P. freyniana may serve as therapeutic agents for RA and cancer treatment.

Synthesis of new hybrid quinazoline compounds as antiproliferative agents for breast and colon cancer treatment

Purpose: To evaluate newly synthesized fuoryl quinazoline derivatives for antitumor efficacy.Methods: Fuoryl quinazoline derivatives were synthesized and the structures of the synthesized compounds were characterized using standard techniques. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) technique was used to assess the anti-proliferative properties of the synthesized derivatives in vitro.Results: All quinazoline compounds displayed cytotoxic activity against breast and colon cancer cell lines to varying degrees. Compound IXa with acetohydrazide moiety was the most effective on MCF7 and HCT116 cell lines, with half-maximal inhibitory concentration (IC50) values of 16.70 and 12.54 μM, respectively.Conclusion: N'-benzylidene-2-((2-(furan-2-yl) quinazolin-4-yl) oxy) acetohydrazide IXa showed the strongest anti-proliferative activity against MCF-7 and HCT116 human cancer cell lines.

BR2 cell penetrating peptide effectively delivers anti-p21Ras scFv to tumor cells with ganglioside expression for therapy of ras-driven tumor

PurposeCell membrane penetrating peptide BR2 can bind with ganglioside and introduce foreign drugs into tumor cells. In this study, we employed BR2 to carry the broad-spectrum anti-p21Ras scFv prepared in our laboratory into ganglioside expressing tumor cells for therapy of ras-driven tumors.MethodsBR2-p21Ras scFv gene was cloned to prokaryotic expression vector and expressed in E. coli BL21, then the fusion protein was purified with HisPur Ni-NTA. The immunoreactivity of the fusion protein with p21Ras was detected by ELISA and western blotting. The membrane-penetrating and immune co-localization with p21Ras of the fusion protein were determined by immunofluorescence. The antitumor activity was investigated using MTT, wound healing, colone formation, and apoptosis assays in vitro.ResultsBR2-p21Ras scFv fusion protein was successfully expressed and purified. We found that the fusion protein could specifically penetrate into human tumor cell lines which express ganglioside including human neuroblastoma cell line SK-N-SH, human colon cancer cell line HCT116 and human glioma cell line U251. After entering tumor cells the fusion protein bonded specifically with p21Ras. In vitro experiments revealed that it could significantly inhibit the proliferation, migration, and colone formation of HCT116, SK-N-SH, and U251 cells and promote the apoptosis of these tumor cells.ConclusionsBR2-p21Ras scFv can penetrate ganglioside expressing tumor cells and inhibit the growth of ras-driven tumor by binding with p21Ras, and producing an inhibitory effect. It is suggested that BR2-p21Ras scFv is a potential ras-driven tumor therapeutic antibody.

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.

Two New Austocystin Analogs from the Marine-Derived Fungus Aspergillus sp. WHUF05236.

Two new austocystin analogs, austocystin P (1) and austocystin Q (2), along with fourteen known compounds (3-16) were isolated from the fermentation extract of Aspergillus sp. WHUF05236. The planar structures of 1 and 2 were elucidated through 1D, 2D NMR and MS analyses. Their absolute configurations were determined by the time-dependent density functional (TDDFT)-ECD calculation. Compounds 3, 11, and 12 exhibited antimicrobial activities against Helicobacter pylori with MIC values ranging from 20.00 to 43.47 μM. Compounds 3, 6, and 7 showed cytotoxicities against the human colon cancer cell lines Hct-116 with IC50 values of 101.79, 65.46, and 36.72 μM, respectively.

Effect of Humantenine on mRNA m6A Modification and Expression in Human Colon Cancer Cell Line HCT116.

Humantenine, an alkaloid isolated from the medicinal herb Gelsemium elegans (Gardner & Chapm.) Benth., has been reported to induce intestinal irritation, but the underlying toxicological mechanisms remain unclear. The object of the present study was to investigate the RNA N6-methyladenosine (m6A) modification and distinct mRNA transcriptome profiles in humantenine-treated HCT116 human colon cancer cells. High-throughput MeRIP-seq and mRNA-seq were performed, and bioinformatic analysis was performed to reveal the role of abnormal RNA m6A modification and mRNA expression in humantenine-induced intestinal cell toxicity. After humantenine treatment of HCT116 cells, 1401 genes were in the overlap of differentially m6A-modified mRNA and differentially expressed mRNA. The Kyoto Encyclopedia of Genes and Genomes and Gene Ontology annotation terms for actin cytoskeleton, tight junctions, and adherens junctions were enriched. A total of 11 kinds of RNA m6A methylation regulators were differentially expressed. The m6A methylation levels of target genes were disordered in the humantenine group. In conclusion, this study suggested that the HCT116 cell injury induced by humantenine was associated with the abnormal mRNA expression of m6A regulators, as well as disordered m6A methylation levels of target genes.

Self-Assembled Alkylated Polyamine Analogs as Supramolecular Anticancer Agents.

Conformationally restrained polyamine analog PG11047 is a well-known drug candidate that modulates polyamine metabolism and inhibits cancer cell growth in a broad spectrum of cancers. Here, we report a structure–activity relationship study of the PG11047 analogs (HPGs) containing alkyl chains of varying length, while keeping the unsaturated spermine backbone unchanged. Synthesis of higher symmetrical homologues was achieved through a synthetic route with fewer steps than the previous route to PG11047. The amphiphilic HPG analogs underwent self-assembly and formed spherically shaped nanoparticles whose size increased with the hydrophobic alkyl group’s increasing chain length. Assessment of the in vitro anticancer activity showed more than an eight-fold increase in the cancer cell inhibition activity of the analogs with longer alkyl chains compared to PG11047 in human colon cancer cell line HCT116, and a more than ten-fold increase in human lung cancer cell line A549. Evaluation of the inhibition of spermine oxidase (SMOX) showed no activity for PG11047, but activity was observed for its higher symmetrical homologues. Comparison with a reference SMOX inhibitor MDL72527 showed nine-fold better activity for the best performing HPG analog.

COMBINING IN VITRO AND IN OVO ASSAYS TO SCREEN FOR ANTI-CANCER AND ANTI-ANGIOGENIC EFFECTS OF THE LEAF EXTRACTS OF Mallotus cumingii Mull.Arg. (EUPHORBIACEAE)

Treatment for cancer is often challenging and various interventions may have detrimental effects. Due to this, the development of less harmful alternatives such as herbal medicine is essential. The present study aims to determine the leaf phytoconstituents present and the bioactivities of Mallotus cumingii Müll.Arg against cancer cells through the utilization of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) assay and anti-angiogenesis through CAM or chorioallantoic membrane assay. The leaf extracts obtained three fractions namely, methanolic crude (MCME) extracts, hexane extracts (MCHE), and ethyl acetate extracts (MCEA), and were tested on HCT-116 (human colorectal cancer cell line) for in vitro cytotoxicity, and blood vessel density and branching through in ovo CAM assay. Phytochemical analysis showed that the M. cumingii fractions contain phenolic compounds, terpenoids, cardiac glycosides, flavonoids, and saponins. For in vitro set-up, MCME of M. cumingii were separated into MCHE and MCEA partitions and were tested against HCT-116 and obtained an IC50 value of < 30 μg/mL, which is deemed active in cytotoxicity. For in ovo set-up, two concentrations of each extract were applied to the duck eggs. Blood vessel density and number of branching points were measured through the ImageJ analysis. All extracts exhibited anti-angiogenic activity, either by decreasing blood vessel density or the number of branching points. Overall, the study demonstrates the potential of M. cumingii as a source of therapeutic agents.

Eucalyptus brevifolia F. Muell and Eucalyptus stricklandii Maiden leaves extracts: HPLC-DAD, GC-MS analysis and in vitro biological activities, combined with the principal component analysis

Natural products always have been relevant for the advancement of new pharmacological products intended to prevent oxidative stress and many diseases since ancient times. Herein, the chemical composition was investigated via chromatographic analysis of varying polarity solvents (petroleum ether, ethyl acetate, methanol and distilled water) from Tunisian Eucalyptus brevifolia F.M and Eucalyptus stricklandii M. leaves. In vitro provision of biological activities through their antioxidant power (ABTS.+), ability to inhibit 15-lipoxygenase (15-LOX), acetylcholinesterase (AChE), xanthine oxidase (XOD), as well as, cytotoxic potency were undertaken. Chemical profiling showed that the methanolic extracts of both species were the richest in total phenolic compounds. GC-MS analysis of the two Eucalyptus species revealed that spathulenol, globulol, epiglobulol, and trans-Z-α-bisabolene-epoxide were the main volatile compounds detected in the different extracts. In addition, the derivatization procedure allowed the identification of ethylene glycol, and D-fructofuranose as very abundant molecules. The HPLC-DAD has enabled the identification of eight new molecules among them icariin (1.1 mg/g dr), and 5-hydroxy-3’-methoxyflavone (7.5 mg/g dr) in the E. stricklandii MeOH and EtOAc extracts. Aqueous E. brevifolia extract possessed the highest anti-AChE with an IC50 (10 µg/mL). Whereas, the MeOH E. stricklandii extract possessed the highest anti 15-LOX with an IC50 (9.7 µg/mL), as well as, the most important percentage of XOD inhibition 51.1%. All extracts revealed an antiproliferative potency against human cancer cell lines HCT-116, MCF-7 and OVCAR, with an inhibition percentage up to 96%. Taken together, both Eucalyptus species have a broad range of potential applications in several fields, such pharmaceutical, medical and food industries as nutraceutical supplements.

Activation of the atypical NF-κB pathway induced by ionizing radiation is not affected by the p53 status.

DNA double-strand breaks induced by ionizing radiation can activate the atypical NF-κB pathway via ATM-mediated phosphorylation of NEMO/IKKγ. We aimed to determine whether the status of p53 influenced the activation of this particular NF-κB pathway. The NF-κB signaling was activated either by irradiation with a single 8 Gy dose or by TNFα cytokine in p53-proficient and p53-deficient variants of HCT116, RKO, and U2-OS human cancer cell lines. To assess pathway activation the kinetics of phosphorylation (Ser32) and proteolytic degradation of IκBα inhibitor and phosphorylation (Ser536) of RelA(p65) NF-κB subunit were analyzed. Though activation of the radiation-induced atypical pathway was delayed and weakened when compared to the cytokine-induced canonical pathway, no significant differences were noted between p53-proficient and p53-deficient variants, which indicated that activation of both NF-κB pathways was not affected by the p53 status. In marked contrast, the presence of p53 significantly affected downstream effects of NF-κB activation, i.e. transcription of NF-κB-dependent genes. However, different patterns of such interference were observed, which indicated gene-specific and cell-specific mechanisms of interactions between NF-κB and p53 at the transcription regulation level.

S-Benzyl cysteine based cyclic dipeptide super hydrogelator: Enhancing efficacy of an anticancer drug via sustainable release.

Peptide-based low molecular weight supramolecular hydrogels hold promising aspects in various fields of application especially in biomaterial and biomedical sciences such as drug delivery, wound healing, tissue engineering, cell proliferation, and so on due to their extreme biocompatibility. Unlike linear peptides, cyclic peptides have more structural rigidity and tolerance to enzymatic degradation and high environmental stability which make them even better candidates for the above-said applications. Herein, a new small cyclic dipeptide (CDP) cyclo-(Leu-S-Bzl-Cys) (P1) consisting of L-leucine and S-benzyl protected L-cysteine was reported which formed a hydrogel at physiological conditions (at 37°C and pH = 7.46). The hydrogel formed from the cyclic dipeptide P1 showed very good tolerance towards environmental parameters such as pH and temperature and was seen to be stable for more than a year without any deformation. The hydrogel was thermoreversible and stable in the pH range 6-12. Mechanical strength of P1 hydrogel was measured by rheology experiments. Atomic force microscopy (AFM) and field emission scanning electron microscopy (FE-SEM) images revealed that, in aqueous solvents, P1 self-assembled into a highly cross-linked nanofibrillar network which immobilized water molecules inside the cages and formed the hydrogel. The self-assembled cyclic dipeptide acquired the antiparallel β-sheet secondary structure, which was evident from CD and Fourier transform infrared (FT-IR) studies. The β-sheet arrangement and formation of amyloid fibrils were further established by ThT binding assay. Furthermore, P1 was able to form a hydrogel in the presence of the anticancer drug 5-fluorouracil (5FU), and sustainable release of the drug from the hydrogel was measured in vitro. The hydrogelator P1 showed almost no cytotoxicity towards the human colorectal cancer cell line HCT116 up to a considerably high concentration and showed potential application in sustainable drug delivery. The co-assembly of 5FU and P1 hydrogel exhibited much better anticancer activity towards the HCT116 cancer cell line than 5FU alone and decreased the IC50 dose of 5FU to a much lower value.

Assessment of Antioxidant, Immunomodulatory Activity of Oxidised Epigallocatechin-3-Gallate (Green Tea Polyphenol) and Its Action on the Main Protease of SARS-CoV-2-An In Vitro and In Silico Approach.

Owing to the instability of Epigallocatechin Gallate (EGCG), it may undergo auto-oxidation and form oxidised products or dimers. In the present study, we aimed to evaluate the therapeutic effects, including antioxidation and immunomodulatory action, of the Oxidised Epigallocatechin Gallate (O-EGCG) as compared to native EGCG and the action of these compounds on main protease (Mpro) docking against SARS-CoV-2. HCT-116 (Human Colon Cancer) cell lines were used to estimate the total antioxidant capacity and lipid peroxidation levels and pro-inflammatory markers (human IL-6, IL-1β, TNF-α). Further, molecular docking analysis was performed by AutoDock and visualised in Discovery studio. Improved antioxidant capacity of O-EGCG was observed, and there was a significant decrease in the inflammatory markers (IL-1β, IL-6, and TNF-α) when O-EGCG was applied as compared to EGCG. The O-EGCG was shown to be strongly associated with the highest docking score and active site residues of IL-1, IL-6, and TNF- α, as well as the Mpro of SARS-CoV-2, according to in silico approach. The in vitro and in silico analyses indicate an improved therapeutic action of the oxidised form of EGCG. The effective inhibitory action of O-EGCG against SARS-CoV-2 suggests further exploration of the compound against COVID-19 and its efficacy. However, in vivo studies and understanding of the mechanism of action of O-EGCG may yield a better opinion on the use of O-EGCG and future human clinical trials.

Antibacterial and anticancer profiling of new benzocaine derivatives: Design, synthesis, and molecular mechanism of action

The need for new chemotherapeutics to overcome development of resistance merits research to discover new agents. Benzocaine derivatives are essential compounds in medicinal chemistry due to their various biological activities including antibacterial and anticancer activities. Therefore, this study focuses on the synthesis of new benzocaine derivatives 3a-e, 6, 7a and 7b, 8, 10-14, and 16a-d and their in vitro evaluation as antibacterial agents against gram +ve and -ve strains and as anticancer agents against HepG-2, HCT-116, and MCF-7 human cancer cell lines. The obtained results demonstrated that thiazolidines 6 and 7b showed higher antibacterial and anticancer activity in comparison with the reference drugs. In addition, 6 and 7b showed high potency as inhibitors toward their biological targets, that is DNA gyrase and human topoisomerase IIα, as compared to the reference standard drugs novobiocin and etoposide, respectively. Molecular docking demonstrated that both compounds could identify the active site of their target enzymes and develop effective binding interactions. Absorption, distribution, metabolism and elimination (ADME) and drug-likeness predictions of both compounds showed that they both have good ADME profiles and no structural alerts that might cause toxicity. Based on this, 6 and 7b could serve as lead compounds for the design of more potent antibacterial and anticancer agents.

Cytotoxic effect of azole compounds bearing trifluorophenyl ring on MCF-7, MDA-MB-231, and HCT-116 cancer cell line

Cancer, a disease defined by rapid proliferation of cells, still remains one of the most feared diseases of the modern world. Many structurally different anti-carcinogenic drugs are used in several tumor types such as bladder, colon, ovary, breast, head and neck, testis, lung and prostate cancer. In this study, cytotoxic effects of the compounds with different linkers (ketone, oxime, alcohol, chlorine) between pyrazole and trifluoromethyl on MCF-7 (human breast adenocarcinoma), MDA-MB-231 (human breast adenocarcinoma), and HCT-116 (human colon cancer) cell lines were investigated. Compound a1 was observed to be the most potent compound with IC50 values of 5.84±0.76, 5.01±0.32, 5.57±0.02 µg.ml-1 against these cell lines, respectively. It was found that all compounds were very effective against all the tested cancer cell lines.

Differentiation inducing factor-1 suppressed the epithelial-mesenchymal transition via degradation of Yes-associated protein

Differentiation-inducing factors (DIFs), produced by Dictyostelium discoideum, show anti-tumor activity via several different signaling molecules, including glycogen synthase kinase-3 (GSK-3) and signal transducer and activator of transcription 3 (STAT3). However, the exact mechanism of action of DIFs is still poorly understood. Therefore, to better understand the action of DIF, we performed DNA microarray analysis and found that the activity of the Hippo signaling pathway may be modified. Therefore, we analyzed mRNA expressions of Hippo signaling pathway target genes. Although the mRNA expression of the target genes including CTGF and Cyr61were elevated by DIF-1, surprisingly the protein expression themselves were reduced. In an attempt to elucidate the mechanism, we found that DIF promoted proteolysis of Yes-associated protein (YAP), a transcriptional co-activator of the Hippo signaling pathway, in the human cervical cell line HeLa and the human colon cancer cell line HCT-116. YAP has also been reported to promote epithelial-to-mesenchymal transition (EMT). Indeed, cell migration, cell invasion and expressions of EMT-related proteins (fibronectin, vimentin and N-cadherin) were reduced by DIF-1. These results suggest that DIF-1 suppresses EMT via degradation of YAP in human cancer cell lines.

Potential anti-tumor activity of 13.56\xa0MHz alternating magnetic hyperthermia and chemotherapy on the induction of apoptosis in human colon cancer cell lines HT29 and HCT116 by up-regulation of Bax, cleaved caspase 3&9, and cleaved PARP proteins

BackgroundThe purpose of the present study was to evaluate the efficacy of chemo-magnetic hyperthermia (MH), a combination of alternating magnetic field (AMF) and superparamagnetic nanoparticles (SPIONs) coated with Polyethylene glycol-Poly(butyl acrylate)-Polyethylene glycol (PEG-PBA-PEG) carrying 5-Fluorouracil (5-Fu), at inducing apoptosis in the human cancer cell lines HT29 and HCT116. This process can be mediated by alterations in the expression of apoptotic effector proteins, including Bax, Bcl-2, cleaved caspase 3&9, and cleaved PARP, which are involved in the intrinsic pathway of apoptosis. For this purpose, the cells were cultured as monolayers. Then both cell lines were treated with 5-Fu/magnetic nanoparticles and magnetic hyperthermia. Finally, the effect of treatment on cancer cells was determined by Western blot analysis and flow cytometry.ResultsOur results showed that combined chemo-magnetic thermotherapy significantly increased the apoptosis in colon cancer cells compared to chemotherapy or hyperthermia alone (P < 0.05). Up-regulation of Bax, cleaved caspase 3&9, and cleaved PARP proteins was indicative of apoptosis induction in cancer cells, which are involved in the intrinsic pathway of apoptosis.ConclusionsThis study demonstrates that localized hyperthermia was able to significantly trigger the 5-Fu release and inhibit cell viability, which, due to the synchronization of hyperthermia and chemotherapy, exacerbated the damage of cancer cells.Graphical

Anti-cancer, antiproliferative activity of active anionic H2O8 oxygen solution on HCT-116 cancer cell

HCT116 cells are adherent epithelial cells derived from the human colorectal carcinoma cell line commonly used to study inflammatory responses in colonic epithelial cells. In this study, it was aimed to evaluate the effects of active anionic H2O8 oxygen solution, which is a very strong antiviral and antimicrobial agent, on HCT-116 human colorectal cancer cell line. Cell viability was determined by MTT analysis. Antiproliferative activity of the anionic H2O8 was investigated on HCT 116 (human colorectal carcinoma) cancer cells. Anionic H2O8 displayed the outstanding activities for MTT test, IC50= 9.44 for 24th hour was calculated as IC50= 11.73 for 48th hour on HCT 116 cell line. It is thought that it can serve as an agent with strong potential to be used in treatment.

Synthesis, characterization and biological evaluation of some 2-arylbenzoxazole acetic acid derivatives as potential anticancer agents

A series of 2-arylbenzoxazole compounds possessing a cytotoxic activity as potential anticancer agents has been synthesised. Oxidative coupling of benzaldehyde with o -aminophenol utilizing lead tetraacetate approach has been used to realize the synthesis of compounds 1 - 11 . The cytotoxicity of 1 - 11 have been screened against breast cancer cell line MCF-7 and human colon cancer cell line HCT-116 utilizing doxorubicin as reference drug. Among these compounds, 2-(3-benzyloxyphenyl)benzoxazole-5-acetic acid 5 and 2-(4-methoxyphenyl)benzoxazol-5-acetic acid 10 , are found to be promising cytotoxic compounds against MCF-7 cell line. In addition, this study shows that the presence of acetic acid group at position 5 of benzoxazole nucleus enhances the activity. Moreover, it is noticed that the presence of oxygen atom directly linked to the phenyl substituent improves activity. The results offer a new benzoxazole based template to design and develop novel antineoplastic agents.

CRISPA: A Non-viral, Transient Cas9 Delivery System Based on Reengineered Anthrax Toxin.

Translating the CRISPR/Cas9 genome editing technology into clinics is still hampered by rather unspecific, unsafe and/or inconvenient approaches for the delivery of its main components - the Cas9 endonuclease and a guide RNA - into cells. Here, we describe the development of a novel transient and non-viral Cas9 delivery strategy based on the translocation machinery of the Bacillus anthracis anthrax toxin, PA (protective antigen). We show that Cas9 variants fused to the N-terminus of the lethal factor or to a hexahistidine tag are shuttled through channels formed by PA into the cytosol of human cells. As proof-of-principle, we applied our new approach, denoted as CRISPA, to knock out lipolysis-stimulated lipoprotein receptor (LSR) in the human colon cancer cell line HCT116 and green-fluorescent protein (GFP) in human embryonic kidney 293T cells stably expressing GFP. Notably, we confirmed that the transporter PA can be adapted to recognize specific host cell-surface receptor proteins and may be optimized for cell type-selective delivery of Cas9. Altogether, CRISPA provides a novel, transient and non-viral way to deliver Cas9 into specific cells. Thus, this system is an additional step towards safe translation of the CRISPR/Cas9 technology into clinics.