ABSTRACT
AIM: To prepare RGD-modified long circulating liposome (LCL) loading matrine (RGD-M-LCL) to improve the tumor-targeting and efficacy of matrine. METHODS: LCL which was prepared with HSPC, cholesterol, DSPE-PEG2000 and DSPE-PEG-MAL was modified with an RGD motif confirmed by high performance liquid chromatography (HPLC). The encapsulation efficiency of RGD-M-LCL was also detected by HPLC. MTT assay was used to examine the effects of RGD-M-LCL on the proliferation of Bcap-37, HT-29 and A375 cells. The percentage of apoptotic cells and morphological changes in Bcap-37 cells treated with RGD-M-LCL were detected by Annexin-V-FITC/PI affinity assay and observed under light microscope, respectively. RESULTS: Spherical or oval single-chamber particles of uniform sizes with little agglutination or adhesion were observed under transmission electronic microscope. The RGD motif was successfully coupled to the DSPE-PEG-MAL on liposomes, as confirmed by HPLC. An encapsulation efficiency of 83.13% was obtained when the drug-lipid molar ratio was 0.1, and the encapsulation efficiency was negatively related to the drug-lipid ratio in the range of 0.1-0.4, and to the duration of storage. We found that, compared with free matrine, RGD-M-LCL had much stronger in vitro activity, leading to anti-proliferative and pro-apoptotic effects against cancer cells (P<0.01). CONCLUSION: RGD-M-LCL, a novel delivery system for anti-cancer drugs, was successfully prepared, and we demonstrated that the use of this material could augment the effects of matrine on cancer cells in vitro.
Subject(s)
Alkaloids/chemistry , Alkaloids/pharmacology , Liposomes/chemistry , Quinolizines/chemistry , Quinolizines/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Cell Line, Tumor , Cell Survival/drug effects , Chromatography, High Pressure Liquid , HT29 Cells , Humans , Oligopeptides/chemistry , Phosphatidylcholines/chemistry , Phosphatidylethanolamines/chemistry , Polyethylene Glycols/chemistry , MatrinesABSTRACT
BACKGROUND: Although gastric cancer (GC) remains the second cause of cancer-related death, useful biomarkers for prognosis are still unavailable. We present here the attempt of mining novel biomarkers for GC prognosis by using serum proteomics. METHODS: Sera from 43 GC patients and 41 controls with gastritis as Group 1 and 11 GC patients as Group 2 was successively detected by Surface Enhanced Laser Desorption/ionization Time of Flight Mass Spectrometry (SELDI-TOF-MS) with Q10 chip. Peaks were acquired by Ciphergen ProteinChip Software 3.2.0 and analyzed by Zhejiang University-ProteinChip Data Analysis System (ZJU-PDAS). CEA level were evaluated by chemiluminescence immunoassay. RESULTS: After median follow-up periods of 33 months, Group 1 with 4 GC patients lost was divided into 20 good-prognosis GC patients (overall survival more than 24 months) and 19 poor-prognosis GC patients (no more than 24 months). The established prognosis pattern consisted of 5 novel prognosis biomarkers with 84.2% sensitivity and 85.0% specificity, which were significantly higher than those of carcinoembryonic antigen (CEA) and TNM stage. We also tested prognosis pattern blindly in Group 2 with 66.7% sensitivity and 80.0% specificity. Moreover, we found that 4474-Da peak elevated significantly in GC and was associated with advanced stage (III+IV) and short survival (p < 0.03). CONCLUSION: We have identified a number of novel biomarkers for prognosis prediction of GC by using SELDI-TOF-MS combined with sophisticated bioinformatics. Particularly, elevated expression of 4474-Da peak showed very promising to be developed into a novel biomarker associated with biologically aggressive features of GC.
Subject(s)
Biomarkers, Tumor/blood , Proteomics/methods , Stomach Neoplasms/blood , Area Under Curve , Carcinoembryonic Antigen/blood , Humans , Luminescence , Neoplasm Staging , Prognosis , Protein Array Analysis , ROC Curve , Reproducibility of Results , Sensitivity and Specificity , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Stomach Neoplasms/genetics , Stomach Neoplasms/pathologyABSTRACT
OBJECTIVE: This study is to determine the effect of the natural product parthenolide, a sesquiterpene lactone isolated from extracts of the herb Tanacetum parthenium, on the proliferation of vascular smooth muscle cells (VSMCs). METHODS: Rat aortic VSMCs were isolated and cultured in vitro, and treated with different concentrations of parthenolide (10, 20 and 30 mumol/L). [(3)H]thymidine incorporation was used as an index of cell proliferation. Cell cycle progression and distribution were determined by flow cytometric analysis. Furthermore, the expression of several regulatory proteins relevant to VSMC proliferation including IkappaBalpha, cyclooxygenase-2 (Cox-2), p21, and p27 was examined to investigate the potential molecular mechanism. RESULTS: Treatment with parthenolide significantly decreased the [(3)H]thymidine incorporation into DNA by 30%~56% relative to control values in a dose-dependent manner (P<0.05). Addition of parthenolide also increased cell population at G(0)/G(1) phase by 19.2%~65.7% (P<0.05) and decreased cell population at S phase by 50.7%~84.8% (P<0.05), which is consistent with its stimulatory effects on p21 and p27. In addition, parthenolide also increased IkappaBalpha expression and reduced Cox-2 expression in a time-dependent manner. CONCLUSION: Our results show that parthenolide significantly inhibits the VSMC proliferation by inducing G(0)/G(1) cell cycle arrest. IkappaBalpha and Cox-2 are likely involved in such inhibitory effect of parthenolide on VSMC proliferation. These findings warrant further investigation on potential therapeutic implications of parthenolide on VSMC proliferation in vivo.
Subject(s)
G1 Phase/physiology , Muscle, Smooth, Vascular/cytology , Muscle, Smooth, Vascular/physiology , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/physiology , Resting Phase, Cell Cycle/physiology , Sesquiterpenes/administration & dosage , Animals , Cell Proliferation/drug effects , Cells, Cultured , G1 Phase/drug effects , Muscle, Smooth, Vascular/drug effects , Myocytes, Smooth Muscle/drug effects , Rats , Rats, Sprague-Dawley , Resting Phase, Cell Cycle/drug effectsABSTRACT
In order to study the influence of pregnancy and hormone on the lymphatic stomata of guinea pig's ovary bursa, the lymphatic stomata of ovary bursa were observed during pregnancy and by using exogenous hormone, including serum gonadotrophin (PMSG) combined with human chorionic gonadotropin (HCG) to promote ovulation and androgen to inhibit the ovary. Then the lymphatic stomata in the inner layer of ovary bursa and their absorption functions were observed with scanning electron microscope (SEM) and trypan blue as a tracer respectively. The lymphatic stomata were also counted by the Elescope computer image processing system. We found that the quantity of tracer absorbed by the lymphatic stomata of ovary bursa in pregnant group was more than that in ovulation-promoted group, and least in the androgen-treated group, which had statistical significance between each couples of them (P < 0.05). The opening area of the lymphatic stomata in pregnant, ovulation-promoted and androgen-treated groups were 189.9 +/- 48.7 micron 2/1000 micron 2, 104.4 +/- 31.2 micron 2/1000 micron 2 and 40.5 +/- 18.7 micron 2/1000 micron 2 respectively, which had remarkable statistical difference between each couples (P < 0.01). Under SEM observation, the secretory granules in ciliated columnar epithelium were less in pregnant group than the other two, while both the secretory granules and the cilium were most active in ovulation-promoted group. The results indicated that both the openings and absorption functions of the lymphatic stomata in guinea pig's ovary bursa could be affected by pregnancy and exogenous homone. There existed relationship between ovary function and the regulation of the lymphatic stomata in ovary bursa.
