Involving Cell Cycle Research Articles

Assessment of the Anticancer Effect of Chlorojanerin Isolated from Centaurothamnus maximus on A549 Lung Cancer Cells.

The goal of this study was to assess the anticancer efficacy of chlorojanerin against various cancer cells. The effects of chlorojanerin on cell cytotoxicity, cell cycle arrest, and cell apoptosis were examined using MTT assay, propidium iodide staining, and FITC Annexin V assay. RT-PCR was employed to determine the expression levels of apoptosis-related genes. Furthermore, docking simulations were utilized to further elucidate the binding preferences of chlorojanerin with Bcl-2. According to MTT assay, chlorojanerin inhibited the proliferation of all tested cells in a dose-dependent manner with a promising effect against A549 lung cancer cells with an IC50 of 10 µM. Cell growth inhibition by chlorojanerin was linked with G2/M phase cell cycle arrest in A549 treated cells. Flow cytometry analysis indicated that the proliferation inhibition effect of chlorojanerin was associated with apoptosis induction in A549 cells. Remarkably, chlorojanerin altered the expression of many genes involved in apoptosis initiation. Moreover, we determined that chlorojanerin fit into the active site of Bcl-2 according to the molecular docking study. Collectively, our results demonstrate that chlorojanerin mediated an anticancer effect involving cell cycle arrest and apoptotic cell death and, therefore, could potentially serve as a therapeutic agent in lung cancer treatment.

The oncogenic properties of the EWSR1::CREM fusion gene are associated with polyamine metabolism

The EWSR1::CREM fusion gene, caused by a chromosomal translocation t(10;22)(p11;q12), has been discovered in divergent malignancies, ranging from low-grade to highly malignant cancers. The translocation gives rise to a chimeric protein, EWSR1::CREM. The molecular mechanisms behind the oncogenic properties of the EWSR1::CREM protein have not previously been systematically characterized. In this study, we performed transcriptional profiling of the melanoma cell line CHL-1, with depletion of endogenous EWSR1::CREM protein using siRNA mediated knockdown. We found that the expression of 712 genes was altered (Log2 fold-change ≥ 2). We performed pathway analysis to identify EWSR1::CREM mediated pathways and cell studies to examine functional differences brought upon by the knockdown. Altered pathways involved cell cycle and proliferation, this was further validated by the cell studies where cell migration was affected as well. Among the target genes with the greatest downregulation, we discovered ODC1—a well-established oncogenic enzyme that can be pharmacologically inhibited and is essential for polyamine synthesis. We found that the main effects seen upon EWSR1::CREM knockdown can be reproduced by directly silencing ODC1 expression. These findings provide novel insights into pathogenesis of tumors harboring a EWSR1::CREM fusion gene, hopefully facilitating the development of novel therapeutic strategies.

Hydroalcoholic extract of Buxus sempervirens shows antiproliferative effect on melanoma, colorectal carcinoma and prostate cancer cells by affecting the autophagic flow.

Buxus sempervirens (European Box, Buxaceae, boxwood) has been used in folk medicine to treat rheumatism, arthritis, fever, malaria and skin ulceration while, in recent years, interest has grown on possible employment of boxwood extracts in cancer therapy. We studied the effect of hydroalcoholic extract from dried leaves of Buxus sempervirens (BSHE) on four human cell lines (BMel melanoma cells, HCT116 colorectal carcinoma cells, PC3 prostate cancer cells, and HS27 skin fibroblasts) to ascertain its possible antineoplastic activity. This extract inhibited proliferation of all cell lines in different degree as shown, after 48h-exposure and MTS assay, by the values of GR50 (normalized growth rate inhibition50) that were 72, 48, 38, and 32μg/mL for HS27, HCT116, PC3 and BMel cells, respectively. At the above GR50 concentrations, 99% of all studied cells remained vital showing accumulation of acidic vesicles in the cytoplasm, mainly around nuclei, whereas a higher extract concentration (125μg/mL) was cytotoxic causing, after 48h-exposure, death of all BMel and HCT116 cells. Immunofluorescence showed microtubule-associated light chain three protein (LC3, a marker for autophagy) to be localized on the above acidic vesicles when cells were treated for 48h with BSHE (GR50 concentrations). Western blot analysis revealed, in all treated cells, a significant increase (2.2-3.3 times at 24h) of LC3II, i.e., the phosphatidylethanolamine conjugate of the cytoplasmic form LC3I that is recruited in autophagosome membranes during autophagy. Such increase was accompanied, in all cell lines treated for 24h or 48h with BSHE, by a significant increment (2.5-3.4 times at 24h) of p62, an autophagic cargo protein undergoing degradation during the autophagic process. Therefore, BSHE appeared to promote autophagic flow with its following blockade and consequent accumulation of autophagosome or autolysosomes. The antiproliferative effects of BSHE also involved cell cycle regulators such as p21 (HS27, BMel and HCT116 cells) and cyclin B1 (HCT116, BMel and PC3 cells) whereas, among apoptosis markers, BSHE only decreased (30%-40% at 48h) the expression of the antiapoptotic protein survivin. It was concluded that BSHE impairs autophagic flow with arrest of proliferation and death in both fibroblasts and cancer cells, being the latter much more sensitive to these effects.

Loss of the crumbs cell polarity complex disrupts epigenetic transcriptional control and cell cycle progression in the developing retina.

The crumbs cell polarity complex plays a crucial role in apical-basal epithelial polarity, cellular adhesion, and morphogenesis. Homozygous variants in human CRB1 result in autosomal recessive Leber congenital amaurosis (LCA) and retinitis pigmentosa (RP), with no established genotype-phenotype correlation. The associated protein complexes have key functions in developmental pathways; however, the underlying disease mechanism remains unclear. Using the oko meduzym289/m289 (crb2a-/- ) zebrafish, we performed integrative transcriptomic (RNA-seq data) and methylomic [reduced representation bisulphite sequencing (RRBS)] analysis of whole retina to identify dysregulated genes and pathways. Delayed retinal cell specification was identified in both the crb2a-/- zebrafish and CRB1 patient-derived retinal organoids, highlighting the dysfunction of cell cycle modulation and epigenetic transcriptional control. Differential DNA methylation analysis revealed novel hypermethylated pathways involving biological adhesion, Hippo, and transforming growth factor β (TGFβ) signalling. By integrating gene expression with DNA methylation using functional epigenetic modules (FEM), we identified six key modules involving cell cycle control and disturbance of TGFβ, bone morphogenetic protein (BMP), Hippo, and SMAD protein signal transduction pathways, revealing significant interactome hotspots relevant to crb2a function and confirming the epigenetic control of gene regulation in early retinal development, which points to a novel mechanism underlying CRB1-retinopathies. © 2023 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Design, synthesis, and biological evaluation of tetrahydroisoquinoline based hydroxamate derivatives as HDAC 6 inhibitors for cancer therapy

A series of novel tetrahydroisoquinoline (THIQ) compounds were synthesized and evaluated as histone deacetylase 6 (HDAC 6) inhibitors. The compounds are designed based on various aromatic groups serving as cap group, tetrahydroisoquinoline is used as a linker, and hydroxamic acid acts as a metal-binding group. A total of thirty-one novel THIQ analogues are designed and synthesized (B1-B31). The structures of the synthesized compounds are confirmed using 1H NMR, 13C NMR, LC-MS, and elemental analysis. The characterized compounds are screened for their inhibition against Class I HDAC8, Class IIa HDAC4 and Class IIb HDAC6. Several of these compounds exhibited potent antiproliferative activities in multiple tumor cell lines. Among the compounds tested B10 and B24 were found to be most potent selective inhibitors of HDAC 6 with IC50 values of 0.3 μM and 0.4 μM respectively. B21 and B24 remarkably blocked colony formation in MCF-7 cancer cells. Inhibition of cancer cell proliferation by B21 and B24 involved cell cycle arrest in G1 phase and apoptotic death of the cancer cells.

Novel insights into molecular mechanisms of vegetative cell cycle and resting cyst formation in Apodileptus cf. visscheri (Alveolata, Ciliophora).

Ciliates usually with big cell sizes, complex morphological structures, and diverse life cycles, are good model organisms for studying cell proliferation regulation of eukaryotes. Up to date, the molecular regulation mechanisms for the vegetative cell cycle and encystment of these ciliates are poorly understood. Here, transcriptomes of Apodileptus cf. visscheri, which has an asexual vegetative cell cycle and is apt to encyst when environmental conditions become unfavorable, were sequenced to enrich our related knowledge. In this study, three replicates were sequenced for each of four cell stages, including initial period of growth, morphogenesis, cell division, and resting cyst. The significant transcription differences, involving cell cycle, biosynthesis, and energy metabolism pathways, were revealed between the resting cyst and vegetative cell cycle. Further investigations showed that the cell cycle pathway was enriched during morphogenesis stage and cell division stage. Compared to the initial period of growth stage, the differentially expressed genes involved in cellular components and molecular function were significantly enriched during cell division stage, while cellular components and biological processes were significantly enriched during morphogenesis stage. These provide novel insights into a comprehensive understanding at the molecular level of the survival and adaptive mechanism of unicellular eukaryotes.

A Multiomic Approach to Reversing Therapy Resistance in Multiple Myeloma Using Paired Ex Vivo Drug Sensitivity Measures and RNA Sequencing Data

A key challenge in multiple myeloma (MM) is evolution of resistance to standard-of-care (SOC) therapies such as proteasome inhibitors (PI), monoclonal antibodies, and novel targeted therapies. We developed a computational framework that identifies gene programs associated with SOC therapy resistance by pairing patient RNAseq data with clinical response and ex vivo drug response to find patient-specific targets to overcome therapy resistance. We hypothesize that there are two types of mechanisms of resistance in MM; dynamic (involving cell cycle) and cytostatic (cell-intrinsic, EMDR, complement, etc.). We inferred the dynamic mechanism of resistance by comparing transcriptomic profiles of therapy-naïve vs refractory MM patients, therapy-resistant vs wildtype cell lines, drug perturbation transcriptomics from LINCS-L1000 and activity-based proteomic profiling of MM cell lines. The cytostatic mechanism of resistance was inferred by associating RNAseq data with ex vivo drug response, where CD138+ cells from MM patient samples were co-cultured with human bone marrow stromal cells and patient plasma in an ex vivo reconstruction of the tumor microenvironment and tested with SOC drugs in 399 MM patient samples. The computational framework begins with the construction of a topological map of MM transcriptome using t-SNE and fuzzy c-means clustering on z-normalized expression of 16,738 genes across 844 MM patient samples, which yielded MM-specific gene programs (clusters of co-expressing genes). Using publicly available ChIP-seq databases like ENCODE and ChEA, we identified regulatory proteins (RPs) upstream of these gene programs. Kinases phosphorylating these RPs were identified from proteomic databases like PhosphoSitePlus and PhosphoPoint; generating a cascading GRN of MM gene programs, their RPs, and kinases, described by a biophysical model representing transcription, translation, and post-translational effects. The model estimates expression of each gene as a function of gene expression of its upstream RPs and kinases. The model for transcriptional regulation for each gene is trained using RNAseq data of 422 patients and validated against the rest of the cohort (422). Figure 1 is a graphical representation of gene-wise Pearson's correlation of model-predicted and actual gene expression represented by the color of each point (gene) on the tSNE. The set of genes that have poor correlation in Figure 1 (blue) statistically significantly (hypergeometric test; p-value <1e-48 and representation factor = 7.27) coincide with genes that are epigenetically regulated (identified by their high variability of chromatin accessibility across MM patients). We implement this computational framework to identify gene programs associated with a dynamic mechanism of PI (frontline therapy in MM) resistance, which yielded a GRN comprising of 218 co-expressing genes, 13 upstream RPs, and 45 kinases (17 druggable by kinase inhibitors). Network analysis of this GRN revealed that FOXM1 as a key therapeutic vulnerability (characterized by the highest betweenness centrality - a hub for flow of information) that can be targeted by inhibiting PLK1; a key regulator of cell cycle that phosphorylates FOXM1 that, in turn regulates PLK1 expression. A statistically significant (unpaired t-test) increase in z-normalized expression of PLK1 was observed in MM patients (n = 233) clinically refractory to CFZ (p-value < 0.0001) and BTZ (p-value = 0.0004) compared to PI-naïve. Ex vivo functional validation in a cohort of 61 MM patient samples shows a statistically significant (unpaired t-test) lowering of Volasertib (PLK1 inhibitor) AUC in MM patients that were clinically refractory to CFZ (p-value = 0.03) and BTZ (p-value = 0.02) compared to PI-naïve (Figure 2). Similarly, we infer cytostatic mechanism of resistance to PIs by pairing ex vivo drug response from 399 MM patients with RNAseq data to identify gene programs that are enriched for PI-resistance and PI-sensitivity using gene set enrichment analysis; which identified PRCK, ROCK, and PLK1 inhibitors as therapeutic targets that can either synergize with PIs or exhibit synthetic lethality. Ongoing studies are focused on identifying novel targets to overcome resistance to DARA. The proposed computational approach can identify novel therapeutic strategies to overcome SOC therapy resistance using patient-specific targeted therapies. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Mammography radiomics features at diagnosis and progression-free survival among patients with breast cancer

BackgroundThe associations between mammographic radiomics and breast cancer clinical endpoints are unclear. We aimed to identify mammographic radiomics features associated with breast cancer prognosis.MethodsNested from a large breast cancer cohort in our institution, we conducted an extreme case-control study consisting of 207 cases with any invasive disease-free survival (iDFS) endpoint <5 years and 207 molecular subtype-matched controls with >5-year iDFS. A total of 632 radiomics features in craniocaudal (CC) and mediolateral oblique (MLO) views were extracted from pre-treatment mammography. Logistic regression was used to identify iDFS-associated features with multiple testing corrections (Benjamini–Hochberg method). In a subsample with RNA-seq data (n = 96), gene set enrichment analysis was employed to identify pathways associated with lead features.ResultsWe identified 15 iDFS-associated features from CC-view yet none from MLO-view. S(1,−1)SumAverg and WavEnLL_s-6 were the lead ones and associated with favourable (OR 0.64, 95% CI 0.42–0.87, P = 0.01) and poor iDFS (OR 1.53, 95% CI 1.31–1.76, P = 0.01), respectively. Both features were associated with eight pathways (primarily involving cell cycle regulation) in tumour but not adjacent normal tissues.ConclusionOur findings suggest mammographic radiomics features are associated with breast cancer iDFS, potentially through pathways involving cell cycle regulation.

Synthesis and screening of novel 4-N-heterocyclic-2-aryl-6,7,8-trimethoxyquinazolines as antiproliferative and tubulin polymerization inhibitors

Colchicine binding site represent a crucial target for the anticancer drug development especially in view of emerging drug resistance from the currently available chemotherapeutics. A total of 16 novel 4-N-heterocyclic-2-aryl-6,7,8-trimethoxyquinazolines were synthesized and screened for antiproliferative and tubulin polymerization inhibition potential. The synthesized compounds were evaluated against MCF-7, HeLa and HT-29 cancer cell lines and normal cell line HEK-293 T. In the series, 2‑aryl group with 4‑bromophenyl substitution displayed IC50 values of 6.37 µM, 17.43 µM, 6.76 µM and 4‑chlorophenyl substitution displayed IC50 values of 2.16 µM, 8.53 µM, 10.42 µM against MCF-7, HELA and HT29 cancer cell lines, respectively. In the mechanistic studies involving cell cycle analysis, apoptosis assay and JC-1 studies, both the lead compounds were found to induce mitochondria mediated apoptosis and lead molecule with 4‑chlorophenyl substitution displayed significant tubulin polymerization inhibition activity. In the computation studies, lead molecule displayed significant binding affinites in the colchicine domain and showed good thermodynamic stability during 100 ns MD simulation studies. 4-N-Heterocyclic-2-aryl-6,7,8-trimethoxyquinazolines showed appreciable drug like characteristics and can be developed as potent anticancer agents.

Fisetin Inhibits Trypsin Activity and Suppresses the Growth of Colorectal Cancer in Vitro and in Vivo

Colorectal cancer (CRC) is a malignant tumor with high incidence and bad prognosis. Therapies, which are more safe and effective, are urgently needed. Trypsin is proved to be crucial to cancer proliferation and migration, therefore, it is possible to control cancers by modulating its activity. Fisetin is a flavone with trypsin inhibition properties that was screened from more than 45 compounds derived from traditional Chinese medicine (TCM). However, the effects and mechanisms of fisetin on CRC have not been well investigated. In this study, we evaluated the effects of fisetin on 2 different CRC cell lines. Fisetin remarkably inhibited CRC cell proliferation and migration, as well as induced cell apoptosis and Go/G1 phase arrest in a dose-dependent manner. Mechanistic studies revealed that these effects were mediated partially through signaling pathways involving cell cycle regulators p21, p27, cyclinD1, and NF kappa B (NF-κB) p65. Administration of fisetin also significantly suppressed the tumor growth in tumor-bearing NOD/Shi-scid-IL2R gamma (null) (NOG) mice that had been inoculated with human HCT116 cells. Fisetin at the given dosage did not induce significant acute or chronic toxicity in rats. These data provide a potential therapeutic strategy for CRC.

Abstract 1895: Characterization of gemcitabine- and paclitaxel-loaded poly(lactic-co-glycolic acid) microparticles as a treatment for pancreatic ductal adenocarcinoma using in vitro models

Abstract This study aims to characterize the in vitro efficacy of gemcitabine- and paclitaxel-loaded poly(lactic-co-glycolic acid) microparticles as a treatment modality for pancreatic ductal adenocarcinoma (PDAC). The goal is the development of the drug-loaded microparticles (MPs) for direct intratumoral injection as a treatment option for patients with borderline resectable or locally advanced tumors. Because the MPs are injected directly into the tumor, a higher dose of the chemotherapeutic should reach the cancer cells and minimal, if any, should enter the patient’s systemic circulation, which would increase efficacy and decrease systemic toxicity compared to systemic drug infusions. Methods: Gemcitabine-loaded MPs (GMPs), paclitaxel-loaded MPs (PMPs), and blank (no drug) MPs (BMPs) were formulated using a water in oil in water (W/O/W) emulsion. The human PDAC cell lines MIA PaCa-2 and PANC-1 were treated with GMPs and PMPs alone, or in combination, for three or six days with BMPs used as the control. After the treatment course, MTS cell viability assays were performed to assess the cytotoxicity of the different treatments, or cells were collected for flow cytometry to analyze apoptosis and cell cycle phase. Cells were also treated with free gemcitabine and paclitaxel to determine the IC50 and compare cytotoxicity to the MPs treatments via cell viability assays and flow cytometry. Additionally, patient derived PDAC organoids were treated with GMPs and PMPs alone, or in combination, for five days with BMPs as the control. After five days, organoid viability was assessed via CellTiter-Glo enabling comparison with free drug treatments. Results: Treatment with the different MPs regimens results in significant cell death in both cell lines and organoids as compared to the controls. MIA PaCa-2 cells were more sensitive to free gemcitabine and GMPs treatment than PANC-1 cells. However, both cell lines were similarly sensitive to free paclitaxel and PMPs treatment, with PANC-1 cells displaying more sensitivity to paclitaxel-based treatments than gemcitabine-based treatments. The combination MPs treatment performed similarly to the GMPs treatment in both cell lines. The treated organoid lines did not have statistically significant differences in their sensitivities to the different MPs treatments. Conclusion and Future Directions: Using in vitro assays, we assessed the effects of the drug-loaded MP treatments on the viability of PDAC cell lines and patient derived organoids. Next steps will involve cell cycle and apoptosis analysis utilizing flow cytometry of the two cell lines after exposure to the MPs treatments. Additionally, the sensitivity of the organoids to MPs treatments will be compared to free drug treatments to assess the efficacy of MPs therapy over traditional therapy. Citation Format: Stanley Soroka, Dennis Plenker, Hardik Patel, Amber Habowski, David Tuveson, Laura Martello-Rooney. Characterization of gemcitabine- and paclitaxel-loaded poly(lactic-co-glycolic acid) microparticles as a treatment for pancreatic ductal adenocarcinoma using in vitro models [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 1895.

G protein-coupled receptor 30 mediates cell proliferation of goat mammary epithelial cells via MEK/ERK&PI3K/AKT signaling pathway

ABSTRACT The mammary gland of mammals possesses the specific function of synthesizing, secreting, and delivering milk. Notably, mammary epithelial cells are considered to be central to control the expansion and remodeling of mammary gland into a milk-secretory organ. And the biological function of mammary gland is mainly regulated by the endocrine system, especially for estrogen. G protein-coupled receptor 30 (GPR30), an estrogen membrane receptor, mediates estrogen-induced functions of physiology and pathophysiology. However, the relationship between estrogen/GPR30 signaling and proliferation of goat mammary epithelial cells (gMECs) is still unclear. Herein, estrogen promoted cell proliferation than control, as evidence by upregulation of cell numbers, BrdU-positive cell counts, and cell viability. Of note, these activities were all obviously reduced by treatment with GPR30 antagonist G15, yet GPR30 agonist G1 increased cell proliferation than control. Further, GPR30 silencing inhibited cell proliferation than negative control. This inhibition was accompanied by a G2/M phase arrest and downregulation of cell cycle regulators. Meanwhile, estrogen increased the phosphorylation of ERK1/2 and AKT. Further, the protein level of p-ERK1/2 and p-AKT was enhanced by GPR30 agonist G1 but inhibited by GPR30 antagonist G15 and GPR30 silencing. Importantly, MEK inhibitor and PI3K inhibitor decreased the expression of cell cycle regulators, and repressed estrogen-induced and G1-driven promotion of cell proliferation, suggesting that estrogen regulated cell proliferation of gMECs through mechanisms involving cell cycle, dependent of GPR30 and MEK/ERK and PI3K/AKT signaling pathway. This may provide a strong theoretical basis for researching estrogen sustained-release drugs promoting breast development and improving lactation performance. Abbreviations: gMECs, goat mammary epithelial cells; E2, 17β-estradiol; GPR30, G protein-coupled receptor 30; shRNA, small hairpin RNA; CDK, cyclin-dependent kinase; PI3K, phosphatidylinositol 3-kinase; AKT, proteinkinase B; MAPK, mitogen-activated protein kinase; MEK, mitogen-activated protein kinase kinase; ERK1/2, extracellular signal-regulated kinase 1/2.

The role of senescence in cellular plasticity: Lessons from regeneration and development and implications for age-related diseases.

Senescence is a cellular state which involves cell cycle arrest and a proinflammatory phenotype, and it has traditionally been associated with cellular and organismal aging. However, increasing evidence suggests key roles in tissue growth and regrowth, especially during development and regeneration. Conversely, cellular plasticity-the capacity of cells to undergo identity change, including differentiation and dedifferentiation-is associated with development and regeneration but is now being investigated in the context of age-related diseases such as Alzheimer disease. Here, we discuss the paradox of the role for cellular senescence in cellular plasticity: senescence can act as a cell-autonomous barrier and a paracrine driver of plasticity. We provide a conceptual framework for integrating recent data and use the interplay between cellular senescence and plasticity to provide insight into age-related diseases. Finally, we argue that age-related diseases can be better deciphered when senescence is recognized as a core mechanism of regeneration and development.

Divergent Signaling Pathways May Lead to Convergence in Cancer Therapy - A Review.

Cancer is a chaos of uncontrolled cell proliferation that has consistently invented new circuitry programs to operate inside the cell machinery. Globally, cancer statistics account for 65% of mortality worldwide, mainly due to the adoption of lifestyle behaviours. In 2020, FDA approved 40 new drugs, out of which 16 (40%) were approved as cancer drugs. Overall, the risk of dying from cancer decreased, but further reductions in cancer death rates can be accelerated by applying existing cancer control knowledge across all the population segments, emphasising those in the lowest socio-economic and other disadvantaged population. Various therapeutic regimes, including low-molecular-weight inhibitors, targeting oncogenic signaling pathways are under development. However, the pitfall of targeted therapies is the quick emergence of acquired drug resistance encumbered with toxic side effects. Several FDA acclaimed therapeutic legacies or biosimilars earmarked signaling pathways of rare diseases (cystic fibrosis, erythropoietic protoporphyria, neuromyelitis optica spectrum disorder, tenosynovial giant cell tumor, sickle cell disease, systemic sclerosis-associated interstitial lung disease, muscular dystrophy), neurological and psychiatric disorders, infectious diseases, heart, lung, circulatory, endocrine diseases, autoimmune conditions, cancers and blood disorders. When cancer progresses, these signals develop specific characteristics that can be targeted for anti-cancer therapy. The designer inhibitors have emerged as novel pharmaceutical interventions that aim to block the pathways in an effort to reverse the abnormal phenotype of the cancer cells. Numerous cell-signaling channels have evolved and invigorated to make off three-dimensional feedback networks. The magnitude of accessible information by pathways occupies curated information as a consortium. To fully appreciate the pivotal roles that signaling cascades play in tumor development, it is necessary to understand the involved signaling cascades in the interaction between cancer cells. The prime endeavour is to canonically curate all signaling pathways involving cell cycle, EGFR, MAPK, GPCR, PI3K/ AKT/mTOR, immune checkpoints, nuclear receptors, janus kinase, transcription activators etc., involving the manipulation of genetic and nuclear receptors. Here, we will summarize the vast amount of information describing the signals that mediate crosstalk between cancer cells and the targets related to this crosstalk.

Pyrogallol-Phloroglucinol-6, 6-Bieckol Restored Primary Cilia Length, Which Was Decreased by High-Fat Diet in Visceral Adipose Tissue, and Decreased Adipogenesis

Length of primary cilia, which involves cell cycle reentry and disassembly of cilia, promotes cell mitosis. It is known that the cilia length in adipose tissue of the high-fat diet (HFD) animals was shortened and accompanied by increased adipogenesis. Male C57BL/6N mice were randomly divided into groups. The mice group was given the normal fat diet (NFD/saline), HFD mice group for 4 weeks, and then HFD was also treated for the next 4 weeks with saline (HFD/saline), Ecklonia cava extract (HFD/ECE), or pyrogallol-phloroglucinol-6, 6-bieckol, a segment of ECE (HFD/PPB). We evaluated the effect of ECE and PPB on modulating cilia length of visceral adipose tissue and decreasing adipogenesis by decreasing cell cycle reentry using an HFD-fed mouse model. ECE and PPB decreased physiological changes, which increased by HFD, but ECE and PPB decreased the upregulation of the IL-6/STAT3/AURKA signaling pathway, which is involved in cilia disassembly. In addition, ECE or PPB elongated the cilia and decreased cyclin A2 and Cdk2 expression, which promote cell cycle reentry, and decreased the adipogenesis genes. PPB and ECE restored cilia length and decreased adipogenesis through modulating the IL-6/STAT3/AURKA pathway and decreasing cell cycle reentry in the visceral adipose tissue of HFD/saline mice group.

Uncovering the mechanisms of dandelion against triple-negative breast cancer using a combined network pharmacology, molecular pharmacology and metabolomics approach

BackgroundTaraxacum mongolicum, also called dandelion, has been used for thousands of years as a remedy for mammary abscess, mammary gland hyperplasia, and various other diseases afflicting the breast. In modern pharmacological research, dandelion has been proven to be effective against triple-negative breast cancer (TNBC). However, the mechanisms of this anti-tumor effect have not been fully elucidated. PurposeThe aim of this investigation was to understand the multi-target mechanisms through which dandelion counteracts TNBC via a network pharmacology strategy as well as to validate its effectiveness by means of molecular pharmacology and metabolomics assessments. MethodsA liquid chromatography coupled with quadrupole time-of-flight mass spectrometer (LC-Q-TOF/MS) was employed to identify the absorbed components of dandelion in rat plasma. The network pharmacology-based prediction was utilized to uncover the potential mechanisms through which dandelion counteracts TNBC, during which potential targets were identified and pathway enrichment analysis was performed. Subsequently, TNBC cells and 4T1 tumor-bearing mice were used to further verify the molecular mechanisms of dandelion. ResultsTwelve active compounds were identified in rat plasma, which were connected with 50 TNBC-related targets. The pathway enrichment showed that dandelion could treat TNBC through regulating a series of biological processes involving cell cycle and metabolism. Experimentally, flow cytometry analysis revealed that dandelion could arrest the G0/G1 and G2/M cell cycles in 4T1 cells. Further western blot analysis evidenced that the protein expression of kinase 6 (CDK6) as well as cyclins B1 and B2 in mice tumor tissue were suppressed by dandelion. In addition, cell metabolomics analysis revealed the changes in the endogenous metabolite levels that result from dandelion treatments, such as the downregulation of arginine and spermine levels. All these findings were consistent with the predicted targets and pathways. ConclusionThis study comprehensively demonstrates the multi-target mechanisms of dandelion against TNBC using network pharmacology, molecular pharmacology, and metabolomics approaches. These findings will provide important stepping stones for further mechanism investigations and may lead to the development of highly effective dandelion-based treatments for TNBC.

Cytotoxic mechanism of tioconazole involves cell cycle arrest at mitosis through inhibition of microtubule assembly.

The online version contains supplementary material available at 10.1007/s10616-021-00516-w.

Tetracaine hydrochloride induces cell cycle arrest in melanoma by downregulating hnRNPA1

Recent evidence suggests potential benefits of applying local anesthetics in cancer patients. Specifically, tetracaine has a potent antitumor effect in diverse cancers, including neuroblastoma, breast cancer, and melanoma; however, the underlying molecular mechanisms remain unclear. Here, we reported that tetracaine hydrochloride inhibited the growth of melanoma cells and arrested melanoma cells in the G0/G1 phase. Tetracaine hydrochloride treatment resulted in translocation of hnRNPA1 from the nucleoplasm to the nuclear envelope and reduced the protein stability of hnRNPA1 possibly by disrupting the dynamic balance of ubiquitination and neddylation. Elevated hnRNPA1 upregulated cyclin D1 to promote cell cycle in melanoma. The hnRNPA1 overexpression attenuated the effect of tetracaine hydrochloride on melanoma cell growth suppression and cell cycle arrest. Furthermore, melanoma homograft experiments demonstrated that tetracaine hydrochloride suppressed melanoma growth, while hnRNPA1 overexpression alleviated tetracaine's antitumor effect on melanoma. Taken together, our findings suggest that tetracaine hydrochloride exerts a potent antitumor effect on melanoma both in vitro and in vivo, and the effect involves cell cycle arrest induction via downregulation of hnRNPA1.

P53 mRNA Metabolism Links with the DNA Damage Response.

Human cells are subjected to continuous challenges by different genotoxic stress attacks. DNA damage leads to erroneous mutations, which can alter the function of oncogenes or tumor suppressors, resulting in cancer development. To circumvent this, cells activate the DNA damage response (DDR), which mainly involves cell cycle regulation and DNA repair processes. The tumor suppressor p53 plays a pivotal role in the DDR by halting the cell cycle and facilitating the DNA repair processes. Various pathways and factors participating in the detection and repair of DNA have been described, including scores of RNA-binding proteins (RBPs) and RNAs. It has become increasingly clear that p53’s role is multitasking, and p53 mRNA regulation plays a prominent part in the DDR. This review is aimed at covering the p53 RNA metabolism linked to the DDR and highlights the recent findings.

Aqueous extract of andean berry induces apoptosis in human colon cancer cells without mitochondrial damage

BACKGROUND: Andean berry contains several classes of phenolic compounds which have showed antioxidant and cytotoxic activity. OBJECTIVE: To characterize the Andean berry aqueous extract and to study their anti-proliferative mechanisms on SW480 and SW620 cell lines (human colon adenocarcinoma). METHODS: Total phenolic and total anthocyanins contents were determined by Folin-Ciocalteau and differential pH methods, respectively. Antioxidant activity was measured by FRAP, ORAC and DPPH methods. Antiproliferative effect was determined by sulforhodamine colorimetric method and apoptosis was analyzed by flow cytometry using propidium iodide/Annexin-V. Mitochondrial potential was evaluated using DIOC6 and ROS levels were measured by 2,7-Dicholorodihydrofluorescein diacetate (DCFH-DA). RESULTS: The total phenol and anthocyanin content were 4409.78± 63,05 mg equivalents of gallic acid/100 mL and 106,57± 1.43 mg equivalents of cyanidin-3-glycoside/100 mL, respectively. Andean berry extract showed antioxidant activity by FRAP, ORAC and DPPH methods and antiproliferative effect on SW480 and SW620 cells. It was observed a cell cycle arrest at S and G2/M phases on SW480 and at G0/G1 phase on SW620 cells. Aqueous extract did not induce mitochondrial depolarization or affect intracellular ROS levels. CONCLUSIONS: Andean berry aqueous extract has antioxidant capacity and induces apoptosis involving cell cycle arrest in SW480 and SW620 cells without mitochondrial damage.