Human Cervical Carcinoma Cell Line Research Articles

Water soluble curcumin with alkyl sulfonate moiety: Synthesis, and anticancer efficacy

Curcumin is classified as a chemotherapeutic medication because of its potential against numerous cancer cell lines and ability to inhibit cancer cell proliferation. Despite these findings, curcumin has yet to be commercialized as a drug due to its low water solubility, low absorption, and restricted bioavailability. As a result, there is a demand for water-soluble curcumin with improved solubility, bioavailability, and thus bioactivity. In this study we report the synthesis and the anticancer activities of water-soluble curcumins derivatives with alkyl sulfonate moiety. The target water-soluble curcumin with alkyl sulfonate moieties was created utilizing a straightforward technique that involved reacting curcumin with various sultones. The cytotoxic (24 h) and cytostatic (72 h) anticancer effect on breast carcinoma (MCF-7), liver carcinoma (HepG2), skin melanoma (B16–F110), colon human cancer and HeLa cervical carcinoma cell lines viability % via MTT assay were determined for the prepared derivatives. Results showed that curcumin-derived compounds have a pronounced cytostatic anticancer effect rather than cytotoxic one in relation to the compound type, cancer cell line type, and examined concentration compared to curcumin. The curcumin sulfonates outperformed curcumin activity against the tested cancer cells and showed to be powerful anticancer candidate drugs as supported by the theoretical calculations. This is evident by their high capacity to form H-bonding during docking with the amino acid side chains and the Vina docking score.

Dissimilar effect of organometallic ruthenium complexes on the viability of MDR and non-MDR experimental models

Ruthenium complexes containing triphenylphosphine diamide ligands were prepared, characterized, and tested for their biological activity against various cancer cell lines and the malaria parasite, Plasmodium falciparum. The effect of M (mono-substituted) and B (bis-substituted) complexes on the human cervical carcinoma (HeLa) cell line was investigated using the MTT assay. Five (B2, B3, B5, B6, and B13) of the 24 synthesized ruthenium complexes showed significant effects with IC50 values ranging between 0.3 and 2.3 μM. Evaluation of the potential biomolecular targets of B2 and B13 by fluorescence spectroscopy revealed relevant interactions with BSA and only a weak affinity for ctDNA. Complexes M2, B2, M13 and B13 were selected for further biological characterization. Their effect on the viability of two ovarian cancer cell lines was compared to normal cell lines, denoting their selectivity. Upon treatment of four different drug-resistant gynaecological cancer cell lines, differing in their multidrug-resistant phenotypes, the efficacy of the bis-substituted complexes was shown to be greater than their mono-substituted counterparts. The non-MDR cells are sensitive to all the tested complexes, compared to MDR cells which are less sensitive. Upon investigation of complexes M2, M13, B2, and B13 against sensitive and multidrug-resistant parasite strains of P. falciparum, the bis-substituted complexes were again shown to be the most potent, with submicromolar activity against both strains. Furthermore, the resistance indexes for the complexes were approximately equal to 1, which is at least 5-fold lower than chloroquine diphosphate, suggesting the ability of these complexes to retain their activity in resistant forms of the parasite.

Glycyrrhizin Impairs the Progression and Metastasis of Cervical Cancer Through the Protein Kinase B/Glycogen Synthase Kinase-3β/β-Catenin Signaling Pathway

The present study assessed the impact of glycyrrhizin on the mechanisms of progression and metastasis in cervical cancer cells. To this end, cell proliferation, migration, and invasion and protein/mRNA expression levels of protein kinase B, glycogen synthase kinase-3 β and β-catenin were measured in three groups of cells: normal cervical epithelial cell line End1/E6E7 (Group A), human cervical squamous cell carcinoma cell line C33a (group B), and human cervical squamous cell carcinoma cell line C33a treated with 50μmol/L glycyrrhizin (Group C). Results showed that glycyrrhizin treatment decreased the proliferation, migration, and invasion of C33a cells and increased apoptosis compared to the cervical cancer control cells. Expression levels of proteins associated with apoptosis such as FS-7-associated surface antigen, Bcl-2-associated X protein and Fas ligand were higher, while protein kinase B pathway activity was attenuated in the glycyrrhizin-treated group compared to the cervical cancer control group. These results suggest that glycyrrhizin may impair cervical cancer cell viability and impede their progression and metastasis by modulating the protein kinase B/glycogen synthase kinase-3β/β-catenin signaling pathway.

Exploring In-Silico, In-Vitro Antioxidant, and Cytotoxic Potential of Valerian wallichii by On Cervical Epithelial Carcinoma (HeLa) Cell Lines.

Traditional medicinal practices often utilize herbal remedies for treating various diseases. This study focuses on exploring the phytochemical constituents, in-silico, in-vitro antioxidant, and anticancer activities of Valerian wallichii root extracts on human cervical epithelial carcinoma (HeLa) cell lines. The molecular docking approach was employed to predict the ligand molecule's orientation within the receptor like Epidermal growth factor receptor tyrosine kinase domain (PDB ID: 1M17) using Schrodinger's GLIDE model. Among the selected phytocompounds, hesperidin exhibited promising inhibitory activity against EGFR (Epidermal Growth Factor Receptor) domain with -8.701 kcal/mol docking score and interactions with MET 769, ASP 831, ASP776, LEU694 and ASN818 residues as compared to standard doxorubicin with -7.6 kcal/mol docking score and interactions with ASP 831, ASN818 and ASP776 residues and further, various antioxidant activity was assessed and In-vitro anticancer activity against HeLa cell lines was evaluated by hydroalcoholic root extracts demonstrated antioxidant capacities, and selective cytotoxicity was observed, with IC50 : 45.759±0.42 μg/mL for the overall extract and IC50 : 30.245±0.58 μg/mL for the EAF fraction as compared to standard doxorubicin with IC50 : 25.9891±0.25 μg/mL, respectively. The present study concluded that Valerian wallichii L possesses potential human cervical epithelial carcinoma cell line inhibition properties based on the computer aided drug design models and in-vitro activity.

Synthesis, spectral characterization and biological evaluation of diosgenin coordinated ruthenium-arene complexes

A set of mononuclear ruthenium complexes, i.e. [Ru(Dg)(C6H6)Cl2] (RuBDg), [Ru(Dg)((p-cymene)Cl2] (RuCDg) were synthesized using a phytosteroid diosgenin (Dg) as ligand. The formulations and characterizations of the complexes were achieved through elemental analyses, infrared, electronic absorption, electron paramagnetic resonances, mass spectrometric analysis, and molar conductance studies. Ruthenium ion is octahedrally coordinated to the oxygen of diosgenin, one arene moiety and two chlorides. An assessment of the antioxidant activities of the compounds indicated better scavenging ability against ABTS•+, DPPH•, O2−, NO• and OH− radicals. The metal complexes showed activity towards free radicals compared to the parent diosgenin. The anticancer activities of diosgenin and the ruthenium complexes were evaluated by MTT assay against the human lung cancer cell line (A549) and cervical carcinoma cell line (HeLa), with cisplatin as a reference. RuCDg exhibited potent ability to kill A549 and HeLa cancer cells with IC50 values of 5.8 ± 0.5 µM and 5.5 ± 0.6 µM, respectively. Similarly, RuBDg showed activity towards A549 and HeLa cancer cells with IC50 values of 16 ± 1 µM and 12 ± 1 µM, respectively. These values are better than that of free diosgenin (A549 − 48 ± 2 µM and HeLa − 43 ± 1 µM). The morphological changes of the HeLa and A549 cells with the complexes, studied by AO-EB and DAPI staining methods, suggested cell death through apoptosis.

The Fatty Acid Composition and the MTT-Cytotoxicity Test of Commercially Available Commiphora Gileadensis Balsam Oil

Commiphora gileadensis, an aromatic plant, is traditionally used in Middle Eastern countries to prevent pain and inflammation. Fatty acid composition, peroxide number and free fatty acidity analyzes of commercially available C. gileadensis balsam oil and investigation of its effect on cell viability. Commercially available C. gileadensis balsam oil fatty acid composition, peroxide number and free fatty acidity values were determined by the IUPAC IID19 method, and MTT cell viability tests were performed on L929 fibroblast and HeLa (human epithelial cervical carcinoma) cell lines. Commercially available C. gileadensis balsam oil did not exert cytotoxic effects on both L929 fibroblast and HeLa (human epithelial cervical carcinoma) cell lines and promoted the growth of cell lines. Due to its growth promoting feature on L929 fibroblast cells, which is a healthy cell line, this material can be used as a cell culture medium additive.

HIV-1 Reverse Transcriptase Expression in HPV16-Infected Epidermoid Carcinoma Cells Alters E6 Expression and Cellular Metabolism, and Induces a Hybrid Epithelial/Mesenchymal Cell Phenotype.

The high incidence of epithelial malignancies in HIV-1 infected individuals is associated with co-infection with oncogenic viruses, such as high-risk human papillomaviruses (HR HPVs), mostly HPV16. The molecular mechanisms underlying the HIV-1-associated increase in epithelial malignancies are not fully understood. A collaboration between HIV-1 and HR HPVs in the malignant transformation of epithelial cells has long been anticipated. Here, we delineated the effects of HIV-1 reverse transcriptase on the in vitro and in vivo properties of HPV16-infected cervical cancer cells. A human cervical carcinoma cell line infected with HPV16 (Ca Ski) was made to express HIV-1 reverse transcriptase (RT) by lentiviral transduction. The levels of the mRNA of the E6 isoforms and of the factors characteristic to the epithelial/mesenchymal transition were assessed by real-time RT-PCR. The parameters of glycolysis and mitochondrial respiration were determined using Seahorse technology. RT expressing Ca Ski subclones were assessed for the capacity to form tumors in nude mice. RT expression increased the expression of the E6*I isoform, modulated the expression of E-CADHERIN and VIMENTIN, indicating the presence of a hybrid epithelial/mesenchymal phenotype, enhanced glycolysis, and inhibited mitochondrial respiration. In addition, the expression of RT induced phenotypic alterations impacting cell motility, clonogenic activity, and the capacity of Ca Ski cells to form tumors in nude mice. These findings suggest that HIV-RT, a multifunctional protein, affects HPV16-induced oncogenesis, which is achieved through modulation of the expression of the E6 oncoprotein. These results highlight a complex interplay between HIV antigens and HPV oncoproteins potentiating the malignant transformation of epithelial cells.

Diastereoselective 1,3-dipolar cycloaddition of cyclopropenes to acenaphthenequinone azomethine ylides

A diastereoselective 1,3-dipolar cycloaddition of azomethine ylides generated from acenaphthylene-1,2-dione (or aceantrylene-1,2-dione) and amino acids with 1,2-diphenylcyclopropenes has been developed. As a result, cyclopropa[a]pyrrolizidines and 3-azabicyclo[3.1.0]hexanes spiro-fused to acenaphthylen-1(2H)-one and aceanthrylen-1(2H)-one fragments were obtained for the first time. Based on DFT calculations at M11/cc-pVDZ level of theory, a concerted mechanism of cycloaddition was proposed to explain the observed diastereoselectivity. The cytotoxic activity of newly synthesized compounds against human erythroleukemia (K562) and human cervical carcinoma (HeLa) cell lines was evaluated in vitro by MTS-assay.

Effect of miR-138 on migration and invasion of cervical cancer cells, and the underlying mechanism

Purpose: To study the influence of microRNA-138 (miR-138) on the migration and invasion of cervical cancer cells, and the underlying mechanism.
 Methods: Fifteen cervical carcinoma subjects were enrolled in the study. Control group comprised cervical epithelial cell line (End1/E6E7) while cervical cancer group was human cervical squamous cell carcinoma cell line c33a. Both were cultured routinely without any treatment. In miR-138 overexpression group, cells were cultured in progeny of human cervical squamous carcinoma cell line c33a infected with miR-138 gene overexpression lentivirus. Expression levels of miR-138 in excised cervical cancer tissues were determined using qPCR. Cell proliferation was determined with CCK8 assay. Immunoblotting was utilized to assay protein expression levels. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine mRNA expression levels, while cell migration and invasion were assessed by Transwell method.
 Results: There was significant down-regulation of miR-138 expression in cervical cancer tissue, relative to nearby tissues (p < 0.05). In miR-138 overexpression group, cell proliferation, number of migrated and invaded cells were significantly reduced, relative to corresponding levels in cervical cancer cells. There were significantly higher expression levels of apoptosis-related proteins FAS, Bax and FasL in miR-138 overexpression group than in cervical cancer cells, while Bcl-2 was significantly downregulated, relative to cervical cancer group (p < 0.05). In cervical cancer cells, mRNA and protein levels of SIRT1 and HIF-1α were significantly up-regulated, relative to corresponding control, but levels of HIF1α and miR-138 were significantly reduced in overexpression group when compared to cervical cancer group (p < 0.05).
 Conclusion: Up-regulating miR-138 in cervical cancer cells reduces HIF-1α through inhibition of SIRT1 signaling, resulting in suppression of multiplication, migration and invasion of cervical cancer cells, while enhancing apoptotic changes.

Development of new cobalt, copper, and zinc complexes of Schiff-base ligands as prospective chemotherapeutic agents

Fluorobenzaldehyde was converted into new, bidentate Schiff-base ligands with nitrogen and sulphur donor sites in two steps. The first step proceeds via Knoevenagel condensation; the product obtained was treated with thiosemicarbazide to yield the ligand L1. The Schiff-base ligand, L2, was synthesized by treating L1 with 4-chlorobenzylchloride in a basic medium. Co (II), Cu (II), and Zn (II) complexes were synthesized with the ligands L1 and L2. All these complexes were characterized by FTIR, UV–Vis, NMR spectroscopy, ESI-mass spectral analysis, magnetic moment, molar conductance, thermal, powder XRD, and electrochemical studies. From these studies, the molecular formulae of the complexes were found to be [M(L)2Cl2], where M = Co (II), Cu (II) and Zn (II) and L = L1 or L2 based on the ligand used in the synthesis. The synthesized complexes were observed to have a high spin and distorted octahedral geometry and were non-electrolytic. The UV absorption titrimetric analysis confirmed that these complexes would bind strongly to CT-DNA through the intercalative mode. The electrophorograms supported the statement that Cu (II) complexes could effectively catalyze the hydrolytic cleavage of CT-DNA and can be considered potent metallonuclease agents. The DPPH assay determined the antioxidant properties of these complexes, and the cupric complexes were observed to possess better antioxidant properties than the standard used ascorbic acid. The cell viability analysis of these complexes showed an elevated control of proliferation against a human breast adenocarcinoma (MCF-7) cancer cell line compared to a human cervical carcinoma (HeLa) cell line. The results exhibited by these compounds were compared with the standard drug, cisplatin. The binding affinity of the Cu (II) metal ion complex towards CT-DNA was high when compared to Co (II) and Zn (II) metal ions. These results are in line with the cytotoxicity studies. Molecular docking studies also hold up the experimental data. Cu (II) complexes using ligand L2 show higher efficiency than those with ligand L1 in all biological assays and can be considered a potent DNA-targeting drug.

Alternaria alternata F3, a Novel Taxol-Producing Endophytic Fungus Isolated from the Fruits of Taxus cuspidata: Isolation, Characterization, Taxol Yield Improvement, and Antitumor Activity.

In this study, a novel taxol-producing endophytic fungus, strain F3, was isolated from the fruits of Taxus cuspidata and identified as Alternaria alternata according to its macroscopic and microscopic traits and sequence analysis of internal transcribed spacer (ITS). The presence of taxol was detected by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) and confirmed by ultra-high-performance liquid chromatography-electrospray coupled to tandem mass spectrometry (UPLC-ESI-MS/MS) and nuclear magnetic resonance (NMR). The fermentation parameters of strain F3 were then optimized for high taxol production. The maximum taxol yield of 195.4µg L-1 by A. alternata F3 was observed in 200-mL yeast peptone dextrose (YPD) broth, at an initial pH value of 6.0, supplemented with 0.1g L-1 sodium acetate, 0.25g L-1 salicylic acid, and 0.00125g L-1 silver nitrate and inoculum size 2%, and incubated at 28°C and 150rpm for 8days, which was 2.12-fold compared with the initial yield of taxol. Also, fungal taxol exhibited antitumor activity towards human lung carcinoma (A549) cell line and human cervical carcinoma (Hela) cell line with IC50 values of 3.98µgmL-1 and 0.35µgmL-1. Overall, this is the first report on taxol-producing endophytic fungus isolated from the fruits of Taxus. This study offers a novel source for the production of taxol for anticancer treatment.

Scaffold-hopping of Linifanib to Design 6-Phenylisoxazolo[3,4-b]pyridin-3-amine Derivatives as FLT3 Inhibitors for Treating Acute Myeloid Leukemia

Background: Acute myeloid leukemia (AML) is the most common type of blood cancer. Fms-like tyrosine kinase 3 (FLT3) is a member of the class III receptor tyrosine kinase family. Overexpression of FLT3 was found in 70-100% of patients with acute myeloid leukaemia. FLT3 internal tandem duplication alteration (ITD) and the tyrosine kinase domain (TKD) are the most common molecular alteration in AML, and FLT3 has become a promising drug target for AML. Objective: A series of 6-phenylisoxazolo[3,4-b]pyridin-3-amine derivatives F1–F15 with amide bonds as FLT3 inhibitors were designed and synthesized in order to find a new lead compound to treat AML. Methods: We designed an original scaffold-hopping protocol by combing the RECAP tool with the Gilde-Based Core-Hopping tool to design novel FLT3 inhibitors based on Linifanib. Inhibitors assembled were ranked by the docking scores generated by Glide. Compounds undisclosed among the top 10 were selected to design a series of 6-phenylisoxazolo[3,4-b]pyridin-3-amine derivatives as FLT3 inhibitors. The kinase inhibitory activities of the fifteen compounds were assayed on FLT3 and FLT3-ITD. The antitumor activities of the structurally modified compounds F1–F15 were evaluated against MOLM-13 and MV4-11, typical FLT3-dependent human AML cells carrying FLT3-ITD mutants and the FLT3-independent human cervical carcinoma cell line HL-60 (harboring wide-type FLT3). Results: Structure–activity relationship (SAR) analysis showed that F14 could inhibit FLT3 and FLT3-ITD by 52% and 45.55%, respectively, at the concentration of 1 ΜF14 exhibited potent activity against FLT3-dependent human acute myeloid leukemia (AML) cell lines, MOLM-13, and MV4-11 (harboring FLT3-ITD mutant) with IC50 values of 2.558 μM and 1.785 μM, respectively. Conclusion: F14 could be used as a novel lead compound to further develop FLT3 inhibitors against AML with FLT3-ITD mutant

Investigation of the Antineoplastic Effects of 2-(4-Chlorophenyl)-13α-Estrone Sulfamate against the HPV16-Positive Human Invasive Cervical Carcinoma Cell Line SiHa.

Cervical carcinoma is one of the most frequent malignant gynecological cancers in women of reproductive age. Because of the poor tolerability of currently available chemotherapeutic agents, efforts have been focused on developing innovative molecules, including steroids, that exert antineoplastic effects with a better safety profile. In addition to their endocrine properties, certain estrogens exhibit additional biological activities, such as antiangiogenic and anticancer effects. Based on previous studies, the antineoplastic properties of 13α-estrone sulfamate derivatives (13AES1-3) were investigated, and the mechanism of action for the most promising compound 13AES3 was explored. Based on their effects on the viability of different human adherent gynecological cancer cells, the SiHa cervical cell line was used for mechanistic experiments. The most active analog 13AES3 was shown to exert considerable proapoptotic effects, as evidenced by a colorimetric caspase-3 assay and fluorescent double staining. It also elicited antimigratory and anti-invasive effects in a concentration-dependent manner, as evidenced by wound healing and Boyden chamber assays, respectively. Regarding their mechanism of action, 13AES derivatives were shown to inhibit tubulin polymerization, and computer simulations provided a possible explanation for the importance of the presence of the chlorophenyl ring on the estrane skeleton. 13AES3 is considered to be the first 13α-estrone derivative with a significant antineoplastic potency against SiHa cancer cells. Therefore, it might serve as a valuable lead molecule for the design of anticancer agents targeting cervical carcinomas.

Diverse Biological Activity of Benzofuroxan/Sterically Hindered Phenols Hybrids.

Combining two pharmacophores in a molecule can lead to useful synergistic effects. Herein, we show hybrid systems that combine sterically hindered phenols with dinitrobenzofuroxan fragments exhibit a broad range of biological activities. The modular assembly of such phenol/benzofuroxan hybrids allows variations in the phenol/benzofuroxan ratio. Interestingly, the antimicrobial activity only appears when at least two benzofuroxan moieties are introduced per phenol. The most potent of the synthesized compounds exhibit high cytotoxicity against human duodenal adenocarcinoma (HuTu 80), human breast adenocarcinoma (MCF-7), and human cervical carcinoma cell lines. This toxicity is associated with the induction of apoptosis via the internal mitochondrial pathway and an increase in ROS production. Encouragingly, the index of selectivity relative to healthy tissues exceeds that for the reference drugs Doxorubicin and Sorafenib. The biostability of the leading compounds in whole mice blood is sufficiently high for their future quantification in biological matrices.

Growth inhibition by fusidic acid in cervical, thyroid, and breast carcinoma cell lines.

We investigated the effects of fusidic acid (FA) on human cervical, thyroid, and breast carcinoma cell lines to determine the potential usefulness of FA in cancer treatment. Six cancer cell lines (cervical cancer: Caski, HeLa; thyroid cancer: 8505C, TPC1; and breast cancer: MCF-7, MDA-MB-231) were treated with FA. Furthermore the changes in cell growth, cell cycle duration, and extent of apoptosis were analyzed. After FA treatment, the cancer cells showed a decrease in growth rate. In the cell death assay, the cell populations were similar in each cell type after treatment with FA, indicating that growth inhibition by FA was not related to the induction of apoptosis. FA induced cell cycle arrest at a dose that inhibited growth rate, which varied in different cell types. G0/G1 phase arrest occurs in breast cancer, S phase arrest in 8505C thyroid cancer, and G2/M phase arrest in cervical cancer. These results indicate that FA reduces growth rates by inducing cell cycle arrest. FA treatment can interfere with cell proliferation by inducing cell cycle arrest in human cervical, thyroid, and breast carcinoma cell lines. Thus, FA can be useful in treating human cervical, thyroid, and breast carcinomas.

Catechin, rutin and quercetin in Quercus mongolica Fisch leaves exert inhibitory effects on multiple cancer cells.

We aimed to identify anti-tumor agents in Quercus mongolica Fisch (QMF). Bioactive compounds in QMF leaves, which were extracted using ethanol as a co-solvent. Five point zero six grams of flavonoids were obtained from 100 g of QMF leaves. Catechin (18.4%), rutin (6.3%), ellagic acid (34.9%), quercetin (5.1%) and kaempferol (20.6%) are the main ingredients of the extracts and were further purified by HPLC. CCK-8 cell proliferation assay showed that catechin and ellagic acid exerted strong inhibitory effects on the proliferation of all cancer cells with lower IC50 values against MCF-7 human breast cancer cell lines, SMMC-7721 human hepatocellular carcinoma cells, HeLa human cervical carcinoma cell lines and SKOV3 human ovarian carcinoma cell lines (p < .05). Catechin, rutin and quercetin induced a higher rate of apoptosis and inhibited all cancer cell proliferation by inducing the G0/G1 phase and G2/M phase arrest (p < .05). However, ellagic acid induced tumor cell death, not through apoptosis and there may be other molecular mechanisms. High levels of catechin and ellagic acid in QMF can be developed as potential drugs to treat different types of cancer cells. PRACTICAL APPLICATIONS: Quercus species have been widely studied because of their antioxidant, anti-inflammatory, antimicrobial, and anti-tumor properties. Bioactive compounds in the leaves of Quercus mongolica Fisch have high levels of catechin and ellagic acid, which exert significant inhibitory properties on the proliferation of various types of cancer cells. Therefore, the bioactive compounds may be potential natural drugs in the prevention of cancer development and progression.

Efficient Capture and Separation of Cancer Cells Using Hyaluronic Acid-Modified Magnetic Beads in a Microfluidic Chip.

The efficient isolation and specific discrimination of circulating tumor cells (CTCs) is expected to provide valuable information for understanding tumor metastasis and play an important role in the treatment of cancer patients. In this study, we developed a novel and rapid method for efficient capture and specific identification of cancer cells using hyaluronic acid (HA)-modified SiO2-coated magnetic beads in a microfluidic chip. First, polyacrylamide-surfaced SiO2-coated magnetic beads (SiO2@MBs) were covalently conjugated with HA, and the created HA-modified SiO2@MBs (HA-SiO2@MBs) display binding specificity to HeLa cells (a human cervical carcinoma cell line) overexpressing CD44 receptors. After incubating the HA-SiO2@MBs with cancer cells for 1 h, the mixture of MBs and cells was drawn into a designed microfluidic channel with two inlets and outlets. Through the formation of lamellar flow, cells specifically bound with the HA-SiO2@MBs can be separated under an external magnetic field with a capture efficiency of up to 92.0%. The developed method is simple, fast, and promising for CTC separation and cancer diagnostics applications.

Generation of 3'-OH terminal-triggered encoding of multicolor fluorescence for simultaneous detection of different DNA glycosylases.

Uracil DNA glycosylase (UDG) and human alkyladenine DNA glycosylase (hAAG) are the important DNA glycosylases for initiating the repair of DNA damage, and the aberrant expression of DNA glycosylases is closely associated with various diseases, such as Parkinson's disease, several cancers, and human immunodeficiency. The simultaneous detection of UDG and hAAG is helpful for the study of early clinical diagnosis. However, the reported methods for multiple DNA glycosylase assay suffer from the application of an expensive single-molecule instrument, labor-tedious magnetic separation, and complicated design. Herein, we develop a simple fluorescence method with only three necessary DNA strands for the selective and sensitive detection of multiple DNA glycosylase activity based on the generation of 3'-OH terminal-triggered encoding of multicolor fluorescence. The method can achieve the detection limits of 5.5 × 10-5 U/mL for UDG and 3.3 × 10-3 U/mL for hAAG, which are lower than those of the reported fluorescence methods. Moreover, it can be further used to detect multiple DNA glycosylases in the human cervical carcinoma cell line (HeLa cells), normal human renal epithelial cells (293T cells), and biological fluid and measure the enzyme kinetic parameters of UDG and hAAG.

Catechol-Based Polymers with High Efficacy in Cytosolic Protein Delivery

Catechol-Based Polymers with High Efficacy in Cytosolic Protein Delivery

Integrating Activity-Guided Strategy and Fingerprint Analysis to Target Potent Cytotoxic Brefeldin A from a Fungal Library of the Medicinal Mangrove Acanthus ilicifolius.

Mangrove-associated fungi are rich sources of novel and bioactive compounds. A total of 102 fungal strains were isolated from the medicinal mangrove Acanthus ilicifolius collected from the South China Sea. Eighty-four independent culturable isolates were identified using a combination of morphological characteristics and internal transcribed spacer (ITS) sequence analyses, of which thirty-seven strains were selected for phylogenetic analysis. The identified fungi belonged to 22 genera within seven taxonomic orders of one phyla, of which four genera Verticillium, Neocosmospora, Valsa, and Pyrenochaeta were first isolated from mangroves. The cytotoxic activity of organic extracts from 55 identified fungi was evaluated against human lung cancer cell lines (A-549), human cervical carcinoma cell lines (HeLa), human hepatoma cells (HepG2), and human acute lymphoblastic leukemia cell lines (Jurkat). The crude extracts of 31 fungi (56.4%) displayed strong cytotoxicity at the concentration of 50 μg/mL. Furthermore, the fungus Penicillium sp. (HS-N-27) still showed strong cytotoxic activity at the concentration of 25 µg/mL. Integrating cytotoxic activity-guided strategy and fingerprint analysis, a well-known natural Golgi-disruptor and Arf-GEFs inhibitor, brefeldin A, was isolated from the target active strain HS-N-27. It displayed potential activity against A549, HeLa and HepG2 cell lines with the IC50 values of 101.2, 171.9 and 239.1 nM, respectively. Therefore, combining activity-guided strategy with fingerprint analysis as a discovery tool will be implemented as a systematic strategy for quick discovery of active compounds.