Phytochemicals targeting glycolysis in colorectal cancer therapy: effects and mechanisms of action.

Colorectal cancer (CRC) is the third most common malignant tumor in the world, and it is prone to recurrence and metastasis during treatment. Aerobic glycolysis is one of the main characteristics of tumor cell metabolism in CRC. Tumor cells rely on glycolysis to rapidly consume glucose and to obtain more lactate and intermediate macromolecular products so as to maintain growth and proliferation. The regulation of the CRC glycolysis pathway is closely associated with several signal transduction pathways and transcription factors including phosphatidylinositol 3-kinases/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR), adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK), hypoxia-inducible factor-1 (HIF-1), myc, and p53. Targeting the glycolytic pathway has become one of the key research aspects in CRC therapy. Many phytochemicals were shown to exert anti-CRC activity by targeting the glycolytic pathway. Here, we review the effects and mechanisms of phytochemicals on CRC glycolytic pathways, providing a new method of drug development.

Green credit and enterprise green operation: Based on the perspective of enterprise green transformation.

This paper uses panel data of listed heavily polluting enterprises from 2007 to 2021, based on the perspective of transformation and upgrading of heavy polluters, innovatively studies the impact of green credit on the green operation of enterprises. At the micro level, the research results of this paper verify the effectiveness of green credit policy on the transformation of green enterprises. It is also found that the two intermediary paths of debt cost and government subsidy play a partial intermediary role in the process of green credit promoting green enterprise transformation and upgrading. Green credit policy also moderates the green transformation of enterprises through debt cost and government subsidies. Based on the research results, this paper puts forward targeted policy suggestions from the aspects of financing constraints, government subsidy policies, enterprise technological innovation and green operation, and provides empirical support for the current expansion of green credit policies in China.

Neogambogic acid suppresses characteristics and growth of colorectal cancer stem cells by inhibition of DLK1 and Wnt/ß-catenin pathway.

PURPOSE: Targeting cancer stem cells (CSCs) may be an efficacious strategy against cancer. We were devoted to exploring the role of neogambogic acid in characteristics and growth of colorectal CSCs. METHODS: SW480 and HCT116 cells were treated with neogambogic acid at different concentrations and transfected with siDLK1 and pcDNA3.1-DLK1 plasmids. The effect of neogambogic acid on the viability of SW480 and HCT116 cells was assessed by MTT assay. Spheroid formation assay was adopted to enrich colorectal CSCs from SW480 and HCT116 cells. The effect of neogambogic acid on colony number, aldehyde dehydrogenase (ALDH) level, apoptosis and cell cycle of SW480 and HCT116 CSCs was detected by colony formation and flow cytometry assays. The expressions of CSC markers, proliferation marker (proliferation nuclear antigen (PCNA)), apoptosis markers (cleaved caspase-3, cleaved caspase-9), Wnt/ß-catenin pathway markers (P-GSK3ß, GSK3ß, ß-catenin and Wnt) and DLK1 were determined by qRT-PCR or Western blot. RESULTS: Neogambogic acid suppressed viability, the spheroid formation ability and the levels of CSC markers in colorectal cancer (CRC) cells, accompanied with inhibition of colony-formation and ALDH level, apoptosis induction and G0/G1 phase arrest. Furthermore, neogambogic acid inhibited expressions of PCNA, P-GSK3ß, P-GSK3ß/GSK3ß, ß-catenin and Wnt, but promoted those of cleaved caspase-3, cleaved caspase-9 and GSK3ß in colorectal CSCs. DLK1 silencing caused opposite results. DLK1 overexpression abrogated the effects of neogambogic acid on colorectal CSCs. CONCLUSION: Neogambogic acid could be an efficacious natural compound targeting colorectal CSCs via inhibition of DLK1 and Wnt/ß-catenin pathway. Thus, neogambogic acid may be an attractive agent against CRC.

LncRNA LINC01315 silencing modulates cancer stem cell properties and epithelial-to-mesenchymal transition in colorectal cancer via miR-484/DLK1 axis.

Long non-coding RNA long intergenic non-protein coding RNA 01315 (LncRNA LINC01315) has been found to be implicated in various cancers, but its role and functions in colorectal cancer (CRC) remain to be addressed. Data on LINC01315 expression in CRC were gathered using bioinformatics analysis, and cancer stem cells (CSCs) were sorted by aldehyde dehydrogenase (ALDH) assay and flow cytometry. Migration, invasion, and stemness of CSCs isolated from CRC cells after transfection were determined by scratch, Transwell, and sphere-formation assays, respectively. Tumor xenograft model was constructed. Target genes and potential-binding sites were predicted using online databases and further confirmed via dual-luciferase reporter assay. Relative factors expressions were determined via quantitative real-time polymerase-chain reaction and Western blot as needed. LINC01315 was high-expressed in CRC and ALDH+ cells. LINC01315 silencing suppressed the migration, invasion, and sphere formation of CRC cells and tumor growth, and downregulated expressions of CSC molecules (ALDH, cluster of difference 44 (CD44), Prominin, and sex determining region Y-box 2 (SOX2)), Zinc Finger E-Box Binding Homeobox 1 (ZEB1) and Vimentin but upregulated E-Cadherin expression. MiR-484 could competitively bind with LINC01315, and LINC01315 silencing promoted miR-484 expression. The level of Delta Like Non-Canonical Notch Ligand 1 (DLK1), the target gene of miR-484, was enhanced by overexpressed LINC01315 yet was suppressed by LINC01315 silencing. Also, DLK1 silencing reversed the effects of downregulated miR-484 on migration, invasion, sphere formation, and CSC molecules expressions in CRC cells. LINC01315 silencing modulated CSC properties and epithelial-to-mesenchymal transition via miR-484/DLK1 axis.

Atractylenolide I inhibits antibiotic-induced dysbiosis of the intestinal microbiome.

BACKGROUND: Atractylenolide I (AT-I) is an active component that is isolated from Rhizoma Atractylodis macrocephalae and it exerts anti-apoptotic, anti-oxidant, and anti-coagulant properties, and has been widely applied in the treatment of cardiovascular and cerebrovascular diseases in China. This study aimed to investigate the effects and possible mechanism of AT-I on intestinal dysbacteriosis in a mouse model. METHODS: Mice dysbacteriosis models were established and treated with AT-I, and the intestinal microbiome of the mice were compared. Using antibiotics-induced bacterial elimination in an intestinal dysbacteriosis-associated xenograft model, the gut microbiota-mediated anti-tumor mechanism was investigated. RESULTS: The intestinal microbiome was changed in the dysbacteriosis mice compared to the control mice, and AT-I could affect the intestinal microbiome of the dysbacteriosis mice. Manipulation of gut bacteria in the intestines of the dysbacteriosis-associated xenograft model further confirmed that the inhibition of tumor progression by AT-I was mediated by the gut microbiota, and that the underlying mechanism involves down-regulation of TLR4/MyD88/NF-κB signaling. AT-I repressed the phosphorylation of p65-NF-κB as well as the downstream cytokines, IL-6 and IL-1ß, in dysbacteriosis mice. CONCLUSIONS: AT-I may inhibit dysbacteriosis by affecting the intestinal microbiome via the regulation of TLR4/MyD88/NF-κB signaling. The present study provides a basis for the application of AT-I as an alternative medication for treating gastrointestinal disorders.

[Determination of three organotin pesticide residues in apples and cabbages by gas chromatography-electron impact/positive chemical ionization mass spectrometry].

A method for the simultaneous determination of fenbutatin oxide, triphenyltin and cyhexatin in apples and cabbages was developed by gas chromatography-mass spectrometry coupled with two different ionization techniques, electron impact (EI) ionization and positive chemical ionization (PCI). At first, the samples were digested by hydrobromic acid, and extracted by acetone-hexane (1:2, v/v). The extracts were derivatized by sodium tetraethylborate as the derivatization reagent, and cleaned up by the Florisil SPE columns. Finally, the samples were analyzed by GC-EI/MS and GC-PCI/MS in selected ion monitoring (SIM) mode. The results showed that good linearities were obtained with correlation coefficients (r2) greater than 0.997 for fenbutatin oxide in the range of 75-500 µg/L in both foods, triphenyltin in the range of 50-1000 µg/L in cabbages and 50-500 µg/L in apples and cyhexatin in the range of 50-1000 µg/L in the two foods. The accuracy was checked at three spiked levels (50, 100 and 200 µg/kg) in cabbages and apples. The limits of detection (LODs, S/N=3) were 0.01-0.05 mg/kg (EI) and 0.01-0.02 mg/kg (PCI), and the limits of quantifications (LOQs, S/N=10) were 0.03-0.16 mg/kg (EI) and 0.02-0.06 mg/kg (PCI). The average recoveries ranged from 59.24%-97.36% (apples) and 50.54%-94.54% (cabbages) in the mode of EI, and the corresponding values were 65.38%-95.86% and 62.56%-90.44% in the mode of PCI. The relative standard deviations (RSDs) were all less than 6.9% (n=6). The PCI method can be used to improve the accuracy in comparing with the EI, in terms of good selectivity and high sensitivity.

[Determination of four kinds of illegal additive residues in sprouts and source beans by high performance liquid chromatography-tandem mass spectrometry].

A reversed-phase high performance liquid chromatography coupled with tandem mass spectrometric (LC-MS/MS) method was developed for the determination of 4-chlorophenoxyacetic acid, 6-benzylaminopurine, enrofloxacin and norfloxacin residues in sprouts and source beans. The sample was extracted by acetonitrile containing 0.1% acetic acid and concentrated. The chromatographic analysis was carried out on a C18 column with methanol and 0.1% formic acid solution as the mobile phases in gradient elution program. The MS analysis was set in electrospray ionization mode and separated to two segments of positive and negative modes. The precursor ions were m/z 189.9, 226.1, 359.9 and 320.1, while the product ions for quantification were m/z 127.0, 91.2, 315.9 and 276.2 for 4-chlorophenoxyacetic acid, 6-benzylaminopurine, enrofloxacin and norfloxacin, respectively. The calibration curves showed good linearity in the range of 5 - 200 microg/L with correlation coefficients more than 0.995. The limits of detection (LODs) were 1 microg/kg and the limits of quantification (LOQs) were 5 microg/kg for the four compounds spiked in mung bean sprouts and mung beans. The recoveries of the four compounds spiked at three levels of 5.0, 10.0 and 20.0 microg/kg ranged from 70% to 91%, with the relative standard deviations (RSDs) less than 14%. The method established is accurate, sensitive, simple, and has considerable advantages in the analysis of the four kinds of illegal additive residues in sprouts and beans simultaneously.

Simultaneous determination of nine types of phthalate residues in commercial milk products using HPLC-ESI-MS-MS.

A multi-residue method was developed for the confirmation and quantitation of nine types of phthalates in milk using high-performance liquid chromatography electrospray ionization tandem mass spectrometry. The samples were extracted with acetonitrile. The analytes were separated using a 0.1% formic acid-methanol system as the mobile phase, and a linear gradient elution program. Mass spectral acquisition was achieved by selectively monitoring the ions in electro-spray ionization mode. Qualitative analysis was based on the retention time and the mass spectrum results, and the quantity was carried out by comparison with the external standard. The mean recoveries for each analyte ranged from 65.2% to 98.3%, with relative standard deviations below 11.2%. The limits of detection were 5∼25 µg/kg, and the limits of quantitation were 17∼83 µg/kg, depending on the compounds. This method has the merits of convenient operation, high sensitivity, and good repeatability, making it an effective method for analysis of phthalates in milk. And the proposed analytical method has been applied to the analysis of phthalates presented in four commercial milk products. The main phthalate residues were DBP and DMP. And the amount of DBP was found to be more than 100 µg/kg in all this milk products.