alloin and Stomach-Neoplasms

To investigate the molecular mechanism underlying the inhibitory effect of aloin on the proliferation and migration of gastric cancer cells.. Human gastric cancer MGC-803 cells treated with 100, 200 and 300 μg/mL aloin were examined for changes in cell viability, proliferation and migration abilities using CCK-8, EdU and Transwell assays. HMGB1 mRNA level in the cells was detected with RT-qPCR, and the protein expressions of HMGB1, cyclin B1, cyclin E1, E-cadherin, MMP-2, MMP-9 and p-STAT3 were determined using Western blotting. JASPAR database was used to predict the binding of STAT3 to HMGB1 promoter. In a BALB/c-Nu mouse model bearing subcutaneous MGC-803 cell xenograft, the effect of intraperitoneal injection of aloin (50 mg/kg) on tumor growth was observed. The protein expressions of HMGB1, cyclin B1, cyclin E1, E-cadherin, MMP-2, MMP-9 and p-STAT3 in the tumor tissue was examined using Western blotting, and tumor metastasis in the liver and lung tissues was detected using HE staining.. Treatment with aloin concentration-dependently inhibited the viability of MGC-803 cells (. Aloin attenuates the proliferation and migration of gastric cancer cells by inhibiting the STAT3/HMGB1 signaling pathway.

Topics: Animals; Cell Proliferation; Cyclin B1; HMGB1 Protein; Humans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Mice; Signal Transduction; STAT3 Transcription Factor; Stomach Neoplasms

This study was designed to evaluate the pharmacological mechanisms of Aloin against gastric cancer (GC) via network pharmacology analysis combined with experimental verification.. Using network pharmacology methods, the potential targets of Aloin and targets related to GC were screened from public databases. The protein-protein interaction (PPI) network, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed to predict the core targets and pathways of Aloin against GC. The expressions of major targets predicted by network pharmacology in normal stomach tissues and GC tissues and their relationships with overall survival of GC were searched in GEPIA, HPA and DriverDBv3 database. The results of network pharmacology analysis were verified by in vitro experiments.. A total of 129 potential targets were retrieved by searching the intersection of Aloin and GC targets. PPI network analysis indicated that 10 targets, including AKT1 and CASP3, were hub genes. GO enrichment analysis involved 93 biological processes, 19 cellular components, and 37 molecular functions. KEGG enrichment analysis indicated that the anti-cancer effect of Aloin was mediated through multiple pathways, such as PI3K-AKT, FoxO and Ras signaling pathway. Among them, the PI3K-AKT signaling pathway, which contained the largest number of enriched genes, may play a greater role in the treatment of GC. The validation of key targets in GEPIA, HPA and DriverDBv3 database showed that the verification results for most core genes were consistent with this study. Then, the results of in vitro experiment indicated that Aloin could inhibit proliferation of NCI-N87 cells and induce cell apoptosis. The results also showed that Aloin could decrease the mRNA and protein expressions of PI3K and AKT, suggesting that Aloin can treat GC by inducing cell apoptosis and regulating the PI3K-AKT signaling pathway.. This study identified the potential targets of Aloin against GC using network pharmacology and in vitro verification, which provided a new understanding of the pharmacological mechanisms of Aloin in treatment of GC.

Topics: Emodin; Humans; Molecular Docking Simulation; Network Pharmacology; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Stomach Neoplasms

To assess the inhibitory effects of aloin on lactate-induced gastric proliferation and migration of cancer cells and explore the underlying molecular mechanism.. Gastric cancer BGC-823 cells were treated with aloin, lactate or the combination of lactate and different doses of aloin. EdU assay was used to detect the proliferation of BGC-823 cells, and colony formation ability of the cells was evaluated with colony forming assay; wound healing and Transwell assays were used to detect the changes in migration ability of the treated cells. The expression levels of cyclin D1, cyclin E1, PCNA, N-cadherin, E-cadherin, MMP-2, MMP-9 and HMGB1 were determined using Western blotting, and ELISA was performed to detect HMGB1 release. HMGB1 expression was knocked down in BGC-823 cells using RNA interference technique, and the effects of HMGB1 knockdown on proliferation and migration of the cells stimulated with lactate for 24 h were examined using EdU and wound healing assays.. Aloin treatment significantly inhibited lactate-induced proliferation of gastric cancer cells, lowered the colony forming ability of lactate-treated cells, and suppressed lactate-induced migration of the cells. Treatment with lactate obviously up-regulated the expressions of cyclin D1, cyclin E1, PCNA, N-cadherin, MMP-2, MMP-9 and HMGB1, which were significantly inhibited by aloin; aloin significantly reversed inhibition of E-cadherin and blocked the release of HMGB1 in lactate-treated cells. BGC-823 cells with HMGB1 knockdown, as compared with the cells transfected with the negative control plasmid, showed significantly lowered proliferation and migration abilities following lactate treatment.. Aloin inhibits lactate-induced proliferation and migration of gastric cancer cells by down-regulating the expression and release of HMGB1 and the expressions of proliferation- and migration-related proteins.

Topics: Cell Line, Tumor; Cell Proliferation; Emodin; Gene Expression Regulation, Neoplastic; HMGB1 Protein; Humans; Lactic Acid; Stomach Neoplasms

Aloin has been reported to have many pharmacological effects including anti-inflammatory, anti-oxidant and anti-tumour activities. However, the precise molecular mechanisms underlying the anti-tumour properties of aloin are yet to be elucidated.. HGC-27 and BGC-823 gastric cancer cells were treated with aloin. EdU and colony formation assays were used to detect the proliferation ability of cells. The migration of cells was detected using wound healing and transwell assays. Western blotting was used to detect the levels of cyclinD1, cyclin E1, MMPs, N-cadherin, E-cadherin and NOX2. The phosphorylation of Akt, mTOR, P70S6K, S6, Src, stat3 and IκBα were also detected by Western blotting. Flow cytometry was used to detect the cell cycle distribution.The location of p65 in cells was determined by using a confocal microscopy assay. The total amounts of ROS present in cells were measured using an ROS assay kit.. Here, we found that aloin inhibited the proliferation and migration of HGC-27 and BGC-823 gastric cancer cells using a combination of EdU, colony formation, wound healing and transwell assays. Further investigations revealed that aloin decreased the protein expression levels of cyclin D1, N-cadherin, and the matrix metalloproteinases (MMP)-2 and MMP-9; increased E-cadherin expression in a dose-dependent manner; inhibited reactive oxygen species (ROS) generation; and mediated the activation of Akt-mTOR, signal transducer and activator of transcription-3 (Stat3), and NF-κB signalling pathways. Our results also indicated that aloin is able to attenuate the expression levels of the two regulatory proteins of nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2), p47. Our findings suggest a potential role for aloin in the prevention of gastric cancer cell proliferation and migration and provide novel insights into the anti-cancer properties of aloin.

Topics: Cell Movement; Cell Proliferation; Emodin; Humans; NADPH Oxidase 2; Reactive Oxygen Species; Signal Transduction; Stomach Neoplasms; Survival Analysis; Tumor Cells, Cultured

To investigate the effect of aloin on apoptosis of human gastric cancer cells and explore the molecular mechanism.. Gastric cancer MKN-28 and HGC-27 cells were cultured routinely in 1640 medium supplemented with 10% fetal bovine serum and 10% non-essential amino acids (for HGC-27 cells) and treated with different concentrations of aloin for different durations. The cell viability, cell nuclear morphology, and apoptotic rate of the cells were detected using CCK-8 assay, DAPI staining and AnnexinV-FITC/PI, respectively; Western blotting was used to detect the expression levels of PARP, procaspase 3 and the phosphorylation of p38, ERK and JNK. The cells were treated with specific inhibitors of p38, ERK and JNK, and the inhibitory effects on these pathways were detected with Western blotting; DAPI staining was used to detect the effects of inhibitors on apoptosis of gastric cancer cells.. Aloin dose-dependently inhibited the viability and induced apoptosis of HGC-27 and MKN-28 cells. Alion treatment obvious enhanced the phosphorylation of p38 and JNK but decreased ERK phosphorylation in the cells. Blocking ERK activation with the ERK inhibitor obviously enhanced aloin-induced cell apoptosis, where inhibiting p38 and JNK activation partly reversed alion-induced apoptosis in the cells.. Aloin induces apoptosis of human gastric cancer cells

Topics: Apoptosis; Caspase 3; Cell Line, Tumor; Cell Survival; Culture Media; Dose-Response Relationship, Drug; Emodin; Fluorescent Dyes; Humans; Indoles; JNK Mitogen-Activated Protein Kinases; MAP Kinase Signaling System; p38 Mitogen-Activated Protein Kinases; Poly(ADP-ribose) Polymerases; Stomach Neoplasms

Gastric cancer is the third leading cause of cancer-associated death worldwide. Although a decrease in its incidence is observed, gastric cancer still poses a major clinical challenge due to poor prognosis and limited treatments. Barbaloin (BBL) is a main medicinal composition of the Chinese traditional medicine aloe vera. BBL has various bioactivities, including anti-oxidant, anti-inflammatory and anti-tumor properties. Polydopamine (pD)-based surface modification is easy to functionalize polymeric nanoparticles (NPs) surfaces with ligands and/or additional polymeric layers. In the present study, BBL-loaded formulations was developed with pD-modified NPs, which was synthesized by polylactide-TPGS (PLA-TPGS) (pD-PLA-TPGS/NPs). And galactosamine (Gal) was conjugated on the prepared NPs (Gal-pD-PLA-TPGS/NPs) for targeting the gastric cancer cells. Here, we found that BBL-loaded Gal-pD-PLA-TPGS/NPs showed the highest cellular uptake efficacy in gastric cancer cells. Gal-pD-PLA-TPGS/NPs more significantly reduced the gastric cancer cell viability. Further, greater apoptosis, autophagy and ROS generation was induced by Gal-pD-PLA-TPGS/NPs in gastric cancer cells. Additionally, compared to the other two NPs, Gal-pD-PLA-TPGS/NPs most markedly decreased ATP levels in gastric cancer cells. In vivo, Gal-pD-PLA-TPGS/NPs were specifically targeted to tumor site. Moreover, Gal-pD-PLA-TPGS/NPs exhibited the most anti-tumor effects, as evidenced by the lowest tumor volume and tumor weight. Of note, there was no significant difference was observed in body and liver weight, as well as the histological changes in major organs isolated from each group of mice. Together, the findings indicated that BBL-loaded Gal-pD-PLA-TPGS/NPs could be targeted to gastric cancer cells to suppress tumor progression without toxicity.

Topics: Animals; Anthracenes; Antineoplastic Agents, Phytogenic; Biological Transport; Cell Line, Tumor; Cell Proliferation; Drug Compounding; Drug Delivery Systems; Epithelial Cells; Galactosamine; Humans; Indoles; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Nanoparticles; Polyesters; Polymers; Stomach Neoplasms; Treatment Outcome; Tumor Burden; Vitamin E; Xenograft Model Antitumor Assays