Colon Adenocarcinoma Cell Line Research Articles

Protosappanin B enhances the chemosensitivity of 5-fluorouracil in colon adenocarcinoma by regulating the LINC00612/microRNA-590-3p/Golgi phosphoprotein 3 axis

Background5-fluorouracil (5-FU) is conventionally used in chemotherapy for colon adenocarcinomas. Acquired resistance of 5-FU remains a clinical challenge in colon cancer, and efforts to develop targeted agents to reduce resistance have not yielded success. Protosappanin B (PSB), the main component of Lignum Sappan extract, is known to exhibit anti-tumor effects. However, whether and how PSB could improve 5-FU resistance in colon cancer have not yet been established. In this study, we aimed to explore the effects and underlying mechanisms of PSB in 5-FU-induced chemoresistance in colon adenocarcinoma.MethodsForty-seven paired colon cancer tissue samples from patients who received 5-FU chemotherapy were collected as clinical samples. Two 5-FU resistant colon cancer cell lines were established for in vitro experiments. Reverse transcription-quantitative PCR (RT-qPCR) was performed to determine the mRNA and microRNA (miRNA) expression levels in colon adenocarcinoma tissues and cell lines. Cell Counting Kit-8 (CCK-8) and flow cytometry assays were performed to evaluate cell proliferation and apoptosis, respectively.ResultsLINC00612 was highly expressed in colon adenocarcinoma samples and 5-FU resistant colon cancer cells. LINC00612 knockdown enhances 5-FU chemosensitivity in 5-FU resistant cells. Notably, PSB treatment attenuated LINC00612 expression in 5-FU resistant colon adenocarcinoma cells. Moreover, PSB treatment reversed the increase in LINC00612-induced 5-FU resistance. Mechanistically, LINC00612 specifically bound to miR-590-3p, which promoted 5-FU resistance in colon adenocarcinoma cells and attenuated the inhibitory effect of LINC00612 on GOLPH3 expression.ConclusionPSB attenuates 5-FU chemoresistance in colon adenocarcinoma by regulating the LINC00612/miRNA-590-3p/GOLPH3 axis.Graphical

Establishment of disulfidptosis-related LncRNA signature as biomarkers in colon adenocarcinoma.

Metabolic reprogramming is a hallmark of cancer and plays a key role in precision oncology treatment. Long non-coding RNAs (lncRNAs) regulate cancer cell behavior, including metabolism. Disulfidptosis, a newly identified form of regulated cell death triggered by glucose starvation, has yet to be fully understood in colon adenocarcinoma (COAD). This study aimed to confirm the existence and role of disulfidptosis in COAD and identify disulfidptosis-related lncRNAs that may be targeted to induce disulfidptosis in COAD. PI and F-actin staining were used to observe disulfidptosis in COAD cell lines. Disulfidptosis-related lncRNAs were identified based on the expression of disulfidptosis-associated genes in the TCGA-COAD database. A four-lncRNA signature for disulfidptosis was established. Subsequently, loss-of-function assays explored the roles of AC013652.1 and MCM3AP-AS1 in disulfidptosis. Disulfidptosis was observed in COAD cells under glucose starvation and could be reversed by agents that prevent disulfide stress, such as dithiothreitol (DTT) and tris-(2-carboxyethyl)-phosphine (TCEP). The prognostic value of disulfidptosis-associated genes in COAD patients was confirmed, with higher expression indicating longer survival. A disulfidptosis-related lncRNA signature comprising four lncRNAs was established based on the expression of these genes. Among these, AC013652.1 and MCM3AP-AS1 predicted worse prognoses. Furthermore, inhibiting AC013652.1 or MCM3AP-AS1 increased disulfidptosis-associated gene expression and cellular death, which could be reversed by DTT and TCEP. This study provides hitherto undocumented evidence of the existence of disulfidptosis and the prognostic value of disulfidptosis-associated genes in COAD. Importantly, we identified lncRNAs AC013652.1 and MCM3AP-AS1, which suppress disulfidptosis and may serve as potential therapeutic targets for COAD.

Nano-sized natural organic matter interacts with bisphenol A and decreases cytotoxicity to human cells

AbstractWhile the toxicity of pollutants has been rather well explored in simple laboratory conditions, there is little knowledge on their real toxicity in natural environments and living organisms because pollutants are often interacting and trapped into organic matter. Because of these interactions, their real concentrations can also be underestimated. Here we studied the nature, intensity, and strength of the interactions between bisphenol A and nano-sized natural organic matter. The bioavailability and toxicity of the complexed bisphenol A were tested with human colon adenocarcinoma cell lines. Results show that that interaction of bisphenol A with organic matter reduces bisphenol A cytotoxicity. Moreover, the bisphenol A-organic matter interaction is weak in the first hour then very stable after 24 h. Once formed, the bisphenol A-organic matter complex escapes detection and, as a consequence, the levels of pollutants in organic-rich media is most probably underestimated. The mechanism of interaction involves hydrophobic and π-stacking forces inside the core of nano-sized organic matter.

Machine learning-derived multi-omics prognostic signature of pyroptosis-related lncRNA with regard to ZKSCAN2-DT and tumor immune infiltration in colorectal cancer.

Colorectal cancer (CRC) has become the most prevalent gastrointestinal malignant tumor, ranking third (10.2%) in incidence and second (9.2%) in death among all malignancies globally. The most common histological subtype of CRC is colon adenocarcinoma (COAD), although the cause of CRC remains unknown, as there are no valid biomarkers. A thorough investigation was used to build a credible biomolecular risk model based on the pyroptosis-associated lncRNAs discovered for COAD prediction. Furthermore, Cibersort and Tumor Immune Dysfunction and Exclusion (TIDE), the methods of exploring tumor immune infiltration, were adopted in our paper to detect the effects of differential lncRNAs on the tumor microenvironment. Finally, quantitative real-time polymerase chain reaction (qPCR), as the approach of exploring expressions, was utilized on four different cell lines. Seven pyroptosis-related lncRNAs have been identified as COAD predictive risk factors. Cox analysis, both univariate and multivariate, revealed that the established signature might serve as a novel independent factor with prognostic meaning in COAD patients. ZKSCAN2-DT was shown to be considerably overexpressed in the COAD cell line when compared to normal human colonic epithelial cells. Furthermore, ssGSEA analysis results revealed that the immune infiltration percentage of most immune cells dropped considerably as ZKSCAN2-DT expression increased, implying that ZKSCAN2-DT may play an important role in COAD immunotherapy. Our research is the first to identify pyroptosis-related lncRNAs connected with COAD patient prognosis and to construct a predictive prognosis signature, directing COAD patient prognosis in therapeutic interventions.

Impact of the thyroid hormone T3 and its nuclear receptor TRα1 on colon cancer stem cell phenotypes and response to chemotherapies

Colorectal cancers (CRCs) are highly heterogeneous and show a hierarchical organization, with cancer stem cells (CSCs) responsible for tumor development, maintenance, and drug resistance. Our previous studies showed the importance of thyroid hormone-dependent signaling on intestinal tumor development and progression through action on stem cells. These results have a translational value, given that the thyroid hormone nuclear receptor TRα1 is upregulated in human CRCs, including in the molecular subtypes associated with CSC features. We used an established spheroid model generated from the human colon adenocarcinoma cell line Caco2 to study the effects of T3 and TRα1 on spheroid formation, growth, and response to conventional chemotherapies. Our results show that T3 treatment and/or increased TRα1 expression in spheroids impaired the response to FOLFIRI and conferred a survival advantage. This was achieved by stimulating drug detoxification pathways and increasing ALDH1A1-expressing cells, including CSCs, within spheroids. These results suggest that clinical evaluation of the thyroid axis and assessing TRα1 levels in CRCs could help to select optimal therapeutic regimens for patients with CRC.Proposed mechanism of action of T3/TRα1 in colon cancer spheroids. In the control condition, TRα1 participates in maintaining homeostatic cell conditions. The presence of T3 in the culture medium activates TRα1 action on target genes, including the drug efflux pumps ABCG2 and ABCB1. In the case of chemotherapy FOLFIRI, the increased expression of ABC transcripts and proteins induced by T3 treatment is responsible for the augmented efflux of 5-FU and Irinotecan from the cancer cells. Taken together, these mechanisms contribute to the decreased efficacy of the chemotherapy and allow cells to escape the treatment. Created with BioRender.com.

Berberis vulgaris L. Root Extract as a Multi-Target Chemopreventive Agent against Colon Cancer Causing Apoptosis in Human Colon Adenocarcinoma Cell Lines.

Berberis vulgaris L. (Berberidaceae) is a shrub that has been widely used in European folk medicine as an anti-inflammatory and antimicrobial agent. The purpose of our study was to elucidate the mechanisms of the chemopreventive action of the plant's methanolic root extract (BVR) against colon cancer cells. Studies were conducted in human colon adenocarcinoma cell lines (LS180 and HT-29) and control colon epithelial CCD841 CoN cells. According to the MTT assay, after 48 h of cell exposure, the IC50 values were as follows: 4.3, 46.1, and 50.2 µg/mL for the LS180, HT-29, and CCD841 CoN cells, respectively, showing the greater sensitivity of the cancer cells to BVR. The Cell Death Detection ELISAPLUS kit demonstrated that BVR induced programmed cell death only against HT-29 cells. Nuclear double staining revealed the great proapoptotic BVR properties in HT-29 cells and subtle effect in LS180 cells. RT-qPCR with the relative quantification method showed significant changes in the expression of genes related to apoptosis in both the LS180 and HT-29 cells. The genes BCL2L1 (126.86-421.43%), BCL2L2 (240-286.02%), CASP3 (177.19-247.83%), and CASP9 (157.99-243.75%) had a significantly elevated expression, while BCL2 (25-52.03%) had a reduced expression compared to the untreated control. Furthermore, in a panel of antioxidant tests, BVR showed positive effects (63.93 ± 0.01, 122.92 ± 0.01, and 220.29 ± 0.02 mg Trolox equivalents (TE)/g in the DPPH•, ABTS•+, and ORAC assays, respectively). In the lipoxygenase (LOX) inhibition test, BVR revealed 62.60 ± 0.87% of enzyme inhibition. The chemical composition of BVR was determined using a UHPLC-UV-CAD-MS/MS analysis and confirmed the presence of several known alkaloids, including berberine, as well as other alkaloids and two derivatives of hydroxycinnamic acid (ferulic and sinapic acid hexosides). The results are very promising and encourage the use of BVR as a comprehensive chemopreventive agent (anti-inflammatory, antioxidant, and pro-apoptotic) in colorectal cancer, and were widely discussed alongside data from the literature.

Genotoxic and antiproliferative properties of Endopleura uchi bark aqueous extract

ABSTRACT The bark extract from Endopleura uchi has been widely used in traditional medicine to treat gynecological-related disorders, diabetes, and dyslipidemias albeit without scientific proof. In addition, E. uchi bark extract safety, especially regarding mutagenic activities, is not known. The aim of this study was to determine the chemical composition, antitumor, and toxicological parameters attributed to an E. uchi bark aqueous extract. The phytochemical constitution was assessed by colorimetric and chromatographic analyzes. The antiproliferative effect was determined using sulforhodamine B (SRB) assay using 4 cancer cell lines. Cytotoxic and genotoxic activities were assessed utilizing MTT and comet assays, respectively, while mutagenicity was determined through micronucleus and Salmonella/microsome assays. The chromatographic analysis detected predominantly the presence of gallic acid and isoquercitrin. The antiproliferative effect was more pronounced in human colon adenocarcinoma (HT-29) and human breast cancer (MCF-7) cell lines. In the MTT assay, the extract presented an IC50 = 39.1 µg/ml and exhibited genotoxic (comet assay) and mutagenic (micronucleus test) activities at 20 and 40 µg/ml in mouse fibroblast cell line (L929) and mutagenicity in the TA102 and TA97a strains in the absence of S9 mix. Data demonstrated that E. uchi bark possesses bioactive compounds which exert cytotoxic and genotoxic effects that might be associated with its antitumor potential. Therefore, E. uchi bark aqueous extract consumption needs to be approached with caution in therapeutic applications.

Antitumoral activity of different Amaryllidaceae alkaloids: In vitro and in silico assays

Ethnopharmacology relevanceThe plants of Amaryllidaceae family, such as Amaryllis belladonna L., have been used as herbal remedies for thousands of years to address various disorders, including diseases that might today be identified as cancer. Aim of the studyThe objective of this work was to evaluate the potential of three Amaryllidaceae alkaloids against four cancer cell lines. Material and methodsThe alkaloids lycorine, 1-O-acetylcaranine, and montanine were evaluated in vitro against colon adenocarcinoma cell line (HCT-116) and breast carcinoma cell lines (MCF-7, MDAMB231, and Hs578T). Computational experiments (target prediction and molecular docking) were conducted to gain a deeper comprehension of possible interactions between these alkaloids and potential targets associated with these tumor cells. ResultsMontanine presented the best results against HCT-116, MDAMB231, and Hs578T cell lines, while lycorine was the most active against MCF-7. In alignment with the target prediction outcomes and existing literature, four potential targets were chosen for the molecular docking analysis: CDK8, EGFR, ER-alpha, and dCK. The docking scores revealed two potential targets for the alkaloids with scores similar to co-crystallized inhibitors and substrates: CDK8 and dCK. A visual analysis of the optimal docked configurations indicates that the alkaloids may interact with some key residues in contrast to the other docked compounds. This observation implies their potential to bind effectively to both targets. ConclusionsIn vitro and in silico results corroborate with data literature suggesting the Amaryllidaceae alkaloids as interesting molecules with antitumoral properties, especially montanine, which showed the best in vitro results against colorectal and breast carcinoma. More studies are necessary to confirm the targets and pharmaceutical potential of montanine against these cancer cell lines.

Abstract 7126: Synergistic combinations and tissue type-specificity of compounds targeting components of the DNA damage response pathway in 2D culture and 3D spheroid cell viability assays

Abstract Poly (ADP-ribose) polymerase (PARP) inhibitors have changed the landscape of treatment for homologous recombination (HR)-deficient cancers. PARP inhibitors are synthetically lethal with deficiencies in genes involved in the HR pathway, such as BRCA2, but resistance against PARP inhibitors is a major problem in the clinic. The use of new synergistic drug combinations targeting different components of the DNA damage response (DDR) pathway may overcome PARP inhibitor resistance. Sixteen small molecule inhibitors acting on different targets in the DDR pathway (including PARP, Polθ and ATR) were profiled in 2D culture and 3D spheroid cell viability assays. Intracellular ATP content was used as an indirect readout of cell number. Compound synergies were determined by profiling combinations of compounds in a 6-by-6 matrix, and quantifying synergistic effects using the Excess over Bliss score [1]. A pair of isogenic BRCA2-deficient and -proficient DLD-1 colon adenocarcinoma cell lines was used as a model of BRCA2-deficient tumors. In 2D cell viability assays, PARP inhibitors and the Polθ inhibitor ART558 were more active in BRCA2-deficient compared to BRCA2-proficient DLD-1 cells, whereas an ATR inhibitor (ceralasertib) was equally potent in both cell lines. Combining olaparib with ART558 in 2D culture and 3D spheroid studies showed synergistic effects in BRCA2-deficient DLD-1 cells. Moreover, olaparib synergized with ceralasertib in both BRCA2-deficient and -proficient DLD-1 cells. Different DDR-targeting inhibitors were profiled in over a hundred cancer cell lines from diverse tumor tissue origin in 2D assays. This showed a relatively high potency for PARP inhibitors in ovarian cancer cell lines compared to cell lines derived from other tissue types, which is consistent with the clinical application of PARP inhibitors for treatment of ovarian cancers. For ATR inhibitors and ART558, kidney and myeloid cancer cell lines were found to be most sensitive, respectively, suggesting potential therapeutic utility of ATR and Polθ inhibitors for treatment of these cancer types. The genomic determinants of the different sensitivities were investigated by comparing compound potencies in the cell viability assays with the mutation status of more than 500 genes involved in cancer or the DDR pathway in the cell lines. Additionally, cancer cell line sensitivity was correlated with the expression levels of more than 19,000 genes in these cell lines to identify novel RNA signatures capable of predicting response to DDR-targeting inhibitors. This study uncovers new synergistic drug combinations that increase the efficiency of PARP inhibitors, and provides new insights into the determinants of cancer cell line sensitivity to compounds targeting different components of the DDR pathway. [1] Liu et al. Stat. Biopharm. Res. 10, 112-22; 2018 Citation Format: Janneke J. Melis, Daphne J. Kluitmans, Jeffrey J. Kooijman, Jelle Dylus, Sabine Wetemans, Suzan Geerdink-Datema, Jeroen A. de Roos, Yvonne Grobben, Guido J. Zaman. Synergistic combinations and tissue type-specificity of compounds targeting components of the DNA damage response pathway in 2D culture and 3D spheroid cell viability assays [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7126.

Enantioselective analysis of the pesticide imazamox after in vitro permeability study in Caco-2 cells.

Imazamox (IMX), a chiral herbicide used in cereals and oilseed crops to control weeds, is commonly sold as a racemic mixture. Its enantiomers, being chiral compounds, may exhibit unique properties when exposed to chiral environments. While IMX enantiomers have been reported to degrade differently in soil and be toxic to some species, their effects on human systems remain poorly understood. This study utilized Caco-2 (human colon adenocarcinoma cell line) cells to assess the in vitro permeability of a racemic mixture of IMX and its isolated enantiomers. Additionally, the study aimed to evaluate whether the metabolite imazamox-O-desmethyl (IMX-D) forms during the permeability process. An enantioselective chromatographic method was developed, fully validated, and the apparent permeability values were obtained. The apparent permeability of rac-IMX, (+)-IMX, and (-)-IMX was determined to be 4.15×10-5 , 5.78×10-5 , and 7.33×10-5 cm s-1 , respectively. These findings suggest that IMX exhibits high intestinal permeability, with an enantioselective absorption for (-)-IMX as compared to (+)-IMX. Finally, the permeability study in Caco-2 cells revealed that the metabolite IMX-D was not generated.

Doxorubicin-induced chemoresistance in Duke's type B colon adenocarcinoma cell line is aggravated in the presence of TGF-β2 through non-apoptotic cell death.

The current challenge in clinical cancer treatment is chemoresistance. Colon cells have inherently higher xenobiotic transporters expression and hence can attain resistance rapidly. Increased levels of TGF-β2 expression in patients have been attributed to cancer progression, aggressiveness, and resistance. To investigate resistance progression, we treated doxorubicin (dox) to HT-29 colon adenocarcinoma cells in the presence or absence of TGF-β2 ligand. After 1, 3-, and 7-day treatment, we investigated cell proliferation, viability, and cytotoxicity by MTT, trypan blue staining, and lactate dehydrogenase enzyme release. The mechanism of cell death was elucidated by hoechst33342 and propidium iodide dual staining and apoptosis assay. The development of resistance was detected by rhodamine123 efflux and P-glycoprotein (P-gp)/MDR1 antibody staining through fluorimetry and flow cytometry. The colony formation ability of the cells was also elucidated. Inhibition of cell proliferation was noted after day 1, while a significant reduction in viability and a significant increase in lactate dehydrogenase release was detected after day 3. Reduction of intracellular rhodamine123 levels was detected after day 3 and was significantly lower in dox with TGF-β2 treatment compared to dox alone. Increased surface P-gp levels after days 3 and 7 were observed in the treated groups. Hoechst33342/propidium iodide staining and apoptosis assay indicated non-apoptotic cell death. The cells treated with TGF-β2 had higher colony formation ability. TGF-β2 expression might play a significant role in the development of chemoresistance to doxorubicin in Duke's type B colon adenocarcinoma cell line, HT-29.

Chemical composition, antioxidant, antimicrobial and antiproliferative activities of the leaf essential oil of Roxburgh’s Cherry (Eugenia roxburghii), an underutilized species

Eugenia roxburghii (Myrtaceae), known as Roxburgh’s Cherry, is a wild edible fruit-producing plant having multiple use in traditional and folk medicines in India and Sri Lanka. In spite of its nutritional and medicinal importance, there is no report on the chemical composition and bioactivities of the essential oil (EO) of this species. In this article, the chemical composition of the leaf EO of E. roxburghii has been reported for the first time along with its antioxidant, antimicrobial and antiproliferative activities. GC–MS analysis of EO led to the identification of 41 compounds with β-caryophyllene (22.32%) as the major constituent. The EO exhibited strong antioxidant capacity in DPPH and ABTS assays with IC50 values of 15.12 ± 0.23 μg/mL and 7.32 ± 0.05 μg/mL, respectively. It also exhibited strong reducing power ability (EC50 = 1.47 ± 0.06 μg/mL). The EO showed highest sensitivity against enteric pathogenic bacteria, Shigella flexneri (ZOI = 24 ± 0.4 mm, MIC = 31.25 μg/mL) validating its ethnomedicinal claim against dysentery. Further, E. roxburghii EO showed significant antiproliferative activity against human colon (HCT-116, IC50 = 64.84 μg/mL) and prostatic adenocarcinoma (PC-3, IC50=70.11 μg/mL) cell lines. The present study suggests that EO can be used as an antiproliferative agent and for the treatment of enteric diseases.

Euphorbia factor L1 inhibited transport channel and energy metabolism in human colon adenocarcinoma cell line Caco-2

Euphorbia factor L1 (EFL1) is a kind of lathyrane-type diterpenoid and is isolated from the medical herb Euphorbia lathyris L. (Euphorbiaceae); it has been reported with the toxicity that causes intestinal irritation, but the underlying mechanisms are still obscure. The objective of this study was to assess the EFL1-induced intestinal cytotoxicity in human colon adenocarcinoma Caco-2 cells. The Caco-2 cells were treated with EFL1, and the intracellular calcium ion concentration, mitochondrial membrane potential (MMP), mitochondrial permeability transition pore (mPTP), adenosine 5′-triphosphate (ATP) content, ATPase activities, TGF-β1 concentration, and transepithelial electrical resistance (TEER) were detected. The interaction between EFL1 and the tight junction proteins Occludin, Claudin-4, Tricellulin, ZO-1, JAM-1, and E-cadherin was simulated by molecular docking. The expression of proteins involved in the energy metabolism, the ion transporters and aquaporins, the tight junction, and the F-actin cytoskeleton were detected by Western blotting and cell immunofluorescence. As a result, EFL1 decreased the intracellular Ca2+, MMP, mPTP, ATP content, and ATPase activities in the Caco-2 cells. The AMPK/SIRT1/PGC-1α signaling pathway, which regulates the energy metabolism, was inhibited. The ion transporters NEH and CFTR, as well as the aquaporins in the Caco-2 cells, were decreased. The tight junction proteins were down-regulated, and the integrity of the intestinal barrier was injured; TGF-β1 was compensatively increased; so, the intestinal permeability was increased and was characterized by decreased TEER. The morphology of the F-actin cytoskeleton was destroyed. These findings indicated that EFL1 caused cytotoxicity in the human intestinal Caco-2 cells through mitochondrial damage, inhibition of the energy metabolism, and suppression of the ion and water molecule transporters, as well as the down-regulation tight junction and cytoskeleton protiens.

Antitumor activity of pegylated human interferon β as monotherapy or in combination with immune checkpoint inhibitors via tumor growth inhibition and dendritic cell activation

Type I interferons (IFN), especially human IFN alpha (IFNα), have been utilized for antitumor therapy for decades. Human interferon beta (IFNβ) is rarely used for cancer treatment, despite advantages over IFNα in biological activities such as tumor growth inhibition and dendritic cell (DC) activation. The utilization of pegylated human IFNβ (PEG-IFNβ), as monotherapy or in combination with immune checkpoint inhibitors (ICIs) was evaluated in this study through in vivo efficacy studies in syngeneic mouse melanoma, non-small cell lung cancer (NSCLC), and colon adenocarcinoma (COAD) models resistant to immune checkpoint inhibitors (ICIs). In vitro comparative study of PEG-IFNβ and pegylated IFNα-2b was performed in terms of tumor growth inhibition against human melanoma, NSCLC and COAD cell lines and activation of human monocyte-derived DCs (MoDCs). Our data demonstrate that the in vivo antitumor effects of PEG-IFNβ are partially attributable to tumor growth-inhibitory effects and DC-activating activities, superior to pegylated IFNα-2b. Our findings suggest that utilizing PEG-IFNβ as an antitumor therapy can enhance the therapeutic effect of ICIs in ICI-resistant tumors by directly inhibiting tumor growth and induction of DC maturation.

In Vitro Cytotoxic Potential of Ethanol Extract of Dictyopteris acrostichoides Against Human Cancer Cells

AbstractEthanol extracts of Caulerpa racemose (Forsskål) J.Agardh, 1873, Dictyopteris acrostichoides (J.Agardh) Bornet, 1885, Halimeda opuntia (Linnaeus) J.V.Lamouroux, 1816, and Polycladia myrica (S.G.Gmelin) Draima, Ballesteros, F.Rousseau & T.Thibaut, 2010, were tested for their cytotoxicity against human hepatoma, human breast adenocarcinoma, and human colon adenocarcinoma cell lines. Dictyopteris acrostichoides displayed cytotoxicity against human hepatoma, human breast adenocarcinoma, and human colon adenocarcinoma with IC50 values of 11.65, 9.28, and 16.86 µg/ml, respectively, in comparison to doxorubicin as a positive control (IC50 5.72, 5.17, and 5.81 µg/ml, respectively). Metabolic profiling of the D. acrostichoides extract characterized seventeen metabolites. In silico analysis indicated 1-(3-oxo-undecyldisulfanyl)-undecan-3-one was the most active epidermal growth factor receptor inhibitor, while 1-(3-oxo-undecyldisulfanyl)-undecan-3-one and di(3-acetoxy-5-undecenyl) disulfide were the most active vascular endothelial growth factor inhibitors. Furthermore, the ethanol extract of D. acrostichoides was tested against epidermal growth factor receptor kinase (IC50 0.11 µg/ml) compared to lapatinib as a positive control (IC50 0.03 µg/ml), and against vascular endothelial growth factor (IC50 0.276 µg/ml) compared to sorafenib as a positive control (IC50 0.049 µg/ml). Graphical Abstract

High expression of ANKRD6 is an indicator for poor prognosis of colon adenocarcinoma

To explore the expression of ANKRD6 in colon adenocarcinoma (COAD) and its implications for prognosis and treatment of COAD. We investigated the differential expression of ANKRD6 in 20 pairs of COAD and adjacent tissues using immunohistochemistry and in colon cancer cell lines and normal colon cells using real-time quantitative PCR. Gene expression matrices, immune infiltration data, and clinical information of 521 COAD patients and 41 normal tissues were obtained from the TCGA database for analyzing the association of ANKRD6 expression with clinicopathological features, gene set enrichment, and immune infiltration in COAD using R software. The prognostic factors of COAD were evaluated using univariate and multivariate Cox regression and Kaplan-Meier analyses, and a prognostic nomogram containing ANKRD6 was constructed. Western blotting, CCK-8 assay, scratch assay and Transwell assay were performed to assess the effects of ANKRD6 knockdown on biological activities of COAD cells. ANKRD6 was highly expressed in COAD cell lines and upregulated in COAD tissues with higher malignancy (P<0.05). Increased ANKRD6 expression was significantly correlated with decreased overall survival and disease-free survival of COAD patients (P<0.05). Univariate and multivariate Cox regression analyses indicated that ANKRD6 expression was significantly correlated with the overall survival rate of COAD patients (P<0.05). The constructed prognostic nomogram showed a C-index value of 0.739 and effectively predicted the 1-, 3-, and 5-year survival of the patients. ANKRD6 expression was positively correlated with immune infiltration of macrophages (P<0.05) and negatively correlated with Th17 cells and other immune cell infiltration (P<0.05). Bioinformatic analysis suggested that ANKRD6 was mainly enriched in Wnt, vascular endothelial growth factor receptor, and calcium signaling pathways in COAD. In cultured COAD cells, ANKRD6 knockdown significantly suppressed cell proliferation, migration, and invasion (P<0.05). High ANKRD6 expression is an indicator for poor prognosis of COAD, and ANKRD6 may serve as a potential biomarker as well as a therapeutic target for diagnosis and treatment of COAD.

Design, Synthesis, and Biological Evaluation of Some Novel Retinoid Derivatives

Design, Synthesis, and Biological Evaluation of Some Novel Retinoid Derivatives

Iron(II)-cyclopentadienyl compounds are cytotoxic against colon adenocarcinoma cell lines: Ethylenebis(diphenylphosphane) vs. triphenylphosphane

Structure-activity studies aiming to understand the role of each coligand in the formulation of new metallodrugs is an important subject. In that frame, six new compounds with general formula [Fe(η5-C5H5)(dppe)(L)][CF3SO3] with L = benzonitriles (1–4) or carbon monoxide (5) and compound [Fe(η5-C5H5)(CO)(PPh3)2][CF3SO3] (6) were synthesized and compared with three other previously reported compounds [Fe(η5-C5H5)(CO)(L)(PPh3)][CF3SO3]. We were particularly interested in assessing the effect of dppe vs. (PPh3 + CO) for this set of compounds. For that, all compounds were tested against two human colon adenocarcinoma cell lines, Colo205 and the refractile Colo320 (expressing ABCB1, an efflux pump causing multidrug resistance), showing IC50 values in the micromolar range. The presence of dppe in the compound's coordination sphere over (PPh3 + CO) allows for more redox stable compounds showing higher cytotoxicity and selectivity, with improved cytotoxicity towards resistant cells that is not related to the inhibition of ABCB1. Further studies with GSH and H2O2 for selected compounds indicated that their antioxidant ability is not probably the main responsible for their cytotoxicity.

Multi-omics profiling reveals cellular pathways and functions regulated by ALDH1B1 in colon cancer cells

Colon cancer is the third leading cause of cancer death globally. Although early screenings and advances in treatments have reduced mortality since 1970, identification of novel targets for therapeutic intervention is needed to address tumor heterogeneity and recurrence. Previous work identified aldehyde dehydrogenase 1B1 (ALDH1B1) as a critical factor in colon tumorigenesis. To investigate further, we utilized a human colon adenocarcinoma cell line (SW480) in which the ALDH1B1 protein expression has been knocked down by 80% via shRNA. Through multi-omics (transcriptomics, proteomics, and untargeted metabolomics) analysis, we identified the impact of ALDH1B1 knocking down (KD) on molecular signatures in colon cancer cells. Suppression of ALDH1B1 expression resulted in 357 differentially expressed genes (DEGs), 191 differentially expressed proteins (DEPs) and 891 differentially altered metabolites (DAMs). Functional annotation and enrichment analyses revealed that: (1) DEGs were enriched in integrin-linked kinase (ILK) signaling and growth and development pathways; (2) DEPs were mainly involved in apoptosis signaling and cellular stress response pathways; and (3) DAMs were associated with biosynthesis, intercellular and second messenger signaling. Collectively, the present study provides new molecular information associated with the cellular functions of ALDH1B1, which helps to direct future investigation of colon cancer.

Identification of phenolic compounds from inflorescences of non-psychoactive Cannabis sativa L. by UHPLC-HRMS and in vitro assessment of the antiproliferative activity against colorectal cancer

Phenolic compounds from Cannabis sativa L. (Cannabaceae family), in particular cannflavins, are known to possess several biological properties. However, their antiproliferative activity, being of great interest from a medicinal chemistry point of view, has not been deeply investigated so far in the literature. In the light of this, the aim of this study was to obtain an enriched fraction of polyphenols (namely PEF) from inflorescences of a non-psychoactive C. sativa (hemp) variety and to evaluate its antiproliferative activity against cancer cells, capitalizing on a new and selective extraction method for hemp polyphenols, followed by preparative flash column chromatography. Untargeted metabolomics, using a new method based on ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS), was applied here for the first time to fully characterize PEF. Then, the main phenolic compounds were quantified by HPLC-UV. The antiproliferative activity of PEF and of the isolated compounds was assessed in vitro for the first time against Caco-2 and SW480 human colon adenocarcinoma cell lines providing promising IC50 values, in comparison with the reference drug used in therapy for this cancer type. Based on these results, PEF can be considered as a new highly potential therapeutic product to be further investigated against colorectal cancer, thanks to the possible synergistic interaction of its compounds.