ABSTRACT
OBJECTIVE: To investigate the mechanism by which Chinese medicine Shengmai Yin (SMY) reverses epithelial-mesenchymal transition (EMT) through lipocalin-2 (LCN2) in nasopharyngeal carcinoma (NPC) cells CNE-2R. METHODS: Morphological changes in EMT in CNE-2R cells were observed under a microscope, and the expressions of EMT markers were detected using quantitative real-time PCR (RT-qPCR) and Western blot assays. Through the Gene Expression Omnibus dataset and text mining, LCN2 was found to be highly related to radiation resistance and EMT in NPC. The expressions of LCN2 and EMT markers following SMY treatment (50 and 100 µ g/mL) were detected by RT-qPCR and Western blot assays in vitro. Cell proliferation, migration, and invasion abilities were measured using colony formation, wound healing, and transwell invasion assays, respectively. The inhibitory effect of SMY in vivo was determined by observing a zebrafish xenograft model with a fluorescent label. RESULTS: The CNE-2R cells showed EMT transition and high expression of LCN2, and the use of SMY (5, 10 and 20 µ g/mL) reduced the expression of LCN2 and reversed the EMT in the CNE-2R cells. Compared to that of the CNE-2R group, the proliferation, migration, and invasion abilities of SMY high-concentration group were weakened (P<0.05). Moreover, SMY mediated tumor growth and metastasis in a dose-dependent manner in a zebrafish xenograft model, which was consistent with the in vitro results. CONCLUSIONS: SMY can reverse the EMT process of CNE-2R cells, which may be related to its inhibition of LCN2 expression. Therefore, LCN2 may be a potential diagnostic marker and therapeutic target in patients with NPC.
Subject(s)
Epithelial-Mesenchymal Transition , Nasopharyngeal Neoplasms , Animals , Humans , Nasopharyngeal Carcinoma/drug therapy , Nasopharyngeal Carcinoma/genetics , Zebrafish , Cell Proliferation , Cell Line, Tumor , Nasopharyngeal Neoplasms/drug therapy , Nasopharyngeal Neoplasms/genetics , Nasopharyngeal Neoplasms/radiotherapy , Cell Movement , Gene Expression Regulation, NeoplasticABSTRACT
Various amounts of 2-((acryloyloxy)ethyl)trimethylammonium chloride were grafted onto chitosan (CS) via redox polymerization method to obtain water-soluble quaternized CS (QCS). The QCS nanoparticles loaded with bovine serum albumin (BSA) were then produced by ionic gelation with tripolyphosphate (TPP) and further covalently cross-linked with genipin. The formation of QCS nanoparticles was optimized as a function of monomer grafting yield, QCS/TPP weight ratio, and QCS/genipin weight ratio by Box-Behnken design and response surface methodology. The results showed that QCS nanoparticles prepared with a grafting yield of 50%, QCS/TPP weight ratio of 7.67, and QCS/genipin weight ratio of 60 had a particle size of 193.68 ± 44.92 nm, polydispersity of 0.232, zeta potential of +23.97 mV and BSA encapsulation efficiency of 46.37 ± 2.89%, which were close to the predicted values from mathematical models. In vitro drug release studies at pH 1.2 and pH 7.4 exhibited that the release rate of BSA was significantly decreased and the release period was significantly prolonged after QCS nanoparticles cross-linking with genipin. Therefore, QCS nanoparticles cross-linked with TPP/genipin dual cross-linkers may be a promising protein drug carrier for a prolonged and sustained delivery.
ABSTRACT
In this study, 2-[(Acryloyloxy)ethyl]trimethylammonium chloride was graft polymerized onto chitosan (CS) to form quaternary ammonium CS (QAC) by using ammonium persulfate as a redox initiator. Alginate (ALG) microspheres loaded with a water-soluble macromolecular model drug, blue dextran (BD), were obtained by corporation of coaxial gas-flow method and ionic gelation process. CS and QAC were then coated on the surfaces of ALG microspheres to generate core/shell structured CS/ALG and QAC/ALG microspheres, respectively. The experiment result showed that QAC/ALG microspheres had a smaller particle size due to the stronger electrostatic interactions between QAC and ALG molecules. In vitro drug release studies at pH 7.4 and pH 9.0 exhibited that the release rate of BD was significantly decreased after ALG microspheres coating with CS and QAC. Moreover, ALG microspheres coated with QAC showed a prolonged release profile for BD at pH 9.0. Therefore, QAC/ALG microspheres may be a promising hydrophilic macromolecular drug carrier for a prolonged and sustained delivery.
