Tumor cellproliferation, infiltration, migration, and neovascularization are known causes of treatment resistance in glioblastoma multiforme (GBM). The purpose of this study was to determine the effect of radiation on the growth characteristics of primary human GBM developed in a nude rat. Primary GBM cells grown from explanted GBM tissues were implanted orthotopically in nude rats. Tumor growth was confirmed by magnetic resonance imaging on day 77 (baseline) after implantation. The rats underwent irradiation to a dose of 50 Gy delivered subcuratively on day 84 postimplantation (n= 8), or underwent no radiation (n= 8). Brain tissues were obtained on day 112 (nonirradiated) or day 133 (irradiated). Immunohistochemistry was performed to determine tumor cell proliferation (Ki-67) and to assess the expression of infiltration marker (matrix metalloproteinase-2, MMP-2) and cell migration marker (CD44). Tumor neovascularization was assessed by microvessel density using von-Willebrand factor (vWF) staining. Magnetic resonance imaging showed well-developed, infiltrative tumors in 11 weeks postimplantation. The proportion of Ki-67-positive cells in tumors undergoing radiation was (71 ± 15)%compared with (25 ± 12)%in the nonirradiated group (P=0.02). The number of MMP-2-positive areas and proportion of CD44-positive cells were also high in tumors receiving radiation, indicating great invasion and infiltration. Microvessel density analysis did not show a significant difference between nonirradiated and irradiated tumors. Taken together, we found that subcurative radiation significantly increased proliferation, invasion, and migration of primary GBM. Our study provides insights into possible mechanisms of treatment resistance fol owing radiation therapy for GBM.

Low-density lipoprotein receptor-related protein 1 (LRP1, also known as CD91), a multifunctional endocytic and cell signaling receptor, is widely expressed on the surface of multiple cell types such as hepatocytes, ifbroblasts, neu-rons, astrocytes, macrophages, smooth muscle cells, and malignant cells. Emerging invitro and invivo evidence demonstrates that LRP1 is critically involved in many processes that drive tumorigenesis and tumor progression. For example, LRP1 not only promotes tumor cell migration and invasion by regulating matrix metalloproteinase (MMP)-2 and MMP-9 expression and functions but also inhibits cell apoptosis by regulating the insulin receptor, the serine/threonine protein kinase signaling pathway, and the expression of Caspase-3. LRP1-mediated phosphorylation of the extracellular signal-regulated kinase pathway and c-jun N-terminal kinase are also involved in tumor cell proliferation and invasion. In addition, LRP1 has been shown to be down-regulated by microRNA-205 and methylation ofLRP1 CpG islands. Furthermore, a novel fusion gene,LRP1-SNRNP25, promotes osteosarcoma cell invasion and migration. Only by understanding the mechanisms of these effects can we develop novel diagnostic and therapeutic strategies for cancers mediated by LRP1.

Mechanisms of inhibition by aspirin of endometrial carcinoma cell proliferation,migration and invasion

Objective:To investigate the effects and mechanisms of aspirin on the proliferation,migration and invasion of endometrial carcinoma HEC-1A cell lines.Methods:HEC-1A cells were cultured to the exponential phase and treated with different concentrations of aspirin (0.625 mmol/ L,1.25 mmol/L,2.5 mmol/L,5 mmol/L and 10 mmol/L) for 24 to 120 hours.Cell proliferation was assessed by methyl thiazolyl tetrazolium (MTT) assay.The migration and invasion of HEC-1A cells were detected by transwell assay.The protein expressions of vascular endothelial growth factor (VEGF) and ascular endothelial growth factor receptor 2 (VEGFR-2) in HEC-1A cells were determined by western blotting.Results:MTT results showed that aspirin inhibited the growth and proliferation of endometrial cancer cells in concentration and time-dependent manner.Aspirin had a significant inhibitory effect on the migration and invasion of HEC-1A cells (P＜0.05).In addition,aspirin obviously suppressed concentration-dependently the expression levels of VEGF and VEGFR-2 (P ＜ 0.05).Conclusion:Aspirin could inhibit the proliferation,migration and invasion of endometrial cancer cells.The anti-tumor mechanism of aspirin might be related to the inhibition of tumor angiogenesis via blocking the VEGF/VEGFR-2 signaling pathway.

Construction,identification and biological feature study of human hepatocellular carcinoma cells stably expressing HBeAg

Objective:To construct a HepG2 cell line which stably expressing Hepatitis B e antigen (HBeAg) and investigate mthe effects of HBeAg on the proliferation,migration and invasion of HepG2 cells.Methods:The lentivirus carrying HBeAg gene was constructed and packaged.HepG2 cells were infected with the lentivirus and screened with puromycin to obtain HepG2 cells which stably expressing HBeAg (HepG2-HBeAg cells).The expression levels of HBeAg mRNA and protein were detected by RT-qPCR and Western blot,respectively.The content of HBeAg secretion in cell supernatant in both HepG2-HBeAg cells and control cells (HepG2-NC cells and HepG2 cells) were detected by IFMA assay.Furthermore,CCK-8 proliferation assay,colony formation assay and transwell migration and invasion assays were conducted to compare the abilities of cell proliferation,migration and invasion,respectively.Results:The expression of HBeAg in the HepG2-HBeAg cell was significantly higher than those in HepG2 cells and HepG2-NC cells.Secreted HBeAg in the supernatant of HepG2-HBeAg cells was 26.33 ± 2.13 PEIU/mL but was undetectable in supernatant of control cells.The proliferation,migration and invasion were all significantly lower in HepG2-HBeAg cells compared to control cells (P＜0.01).Conclusion:A HepG2 cell line which stably expressing HBeAg was constructed successfully,and the over-expression of HBeAg could attenuate the proliferation,migration and invasion of hepatoeellular carcinoma cells.

Objective:The aim of this study was to investigate the effects of plumbagin (PL), a naphthoquinone derived from the medicinal plant plumbago zeylanica, on the invasion and migration of human breast cancer cells. Methods:Human breast cancer MDA-MB-231SArfp cells were treated with different concentrations of plum-bagin for 24 h. The effects of plumbagin on the migration and invasion were observed by a transwell method. The expressions of IL-1α, IL-1β, IL-6, IL-8, TGF-β, TNFα, MMP-2 and MMP-9 mRNA in M DA-MB-231SArfp cells were detected using Real-Time PCR. MDA-MB-231SArfp cells were treated with plumbagin at different concentrations for 45 minutes. The activation of STAT3 was detected by western blot. Following this analysis, STAT3 in MDA-MB-231SArfp cells was knocked out using specific siRNA. mRNA levels of IL-1α, TGF-β, MMP-2 and MMP-9 were then detected. Consequently, MDA-MB-231SArfp cells were injected intracardially into BALB/c nude mice to construct a breast cancer bone metastatic model. The mice were injected intra-peritoneally with plumbagin. Non-invasive in vivo monitoring, X-ray imaging and histological staining were performed to investigate the effects of plumbagin on the invasion and migration of breast cancer cells in vivo. Results: The in vitro results showed that plumbagin could suppress the migration and invasion of breast cancer cells and down-regulate mRNA expressions of IL-1α, TGF-β, MMP-2 and MMP-9. Western blotting demonstrated that plumbagin inhibited the activation of STAT3 signaling in MDA-MB-231SArfp cells. The inactivation of STAT3 was found to have an inhibitory effect on the expressions of IL-1α, TGF-β, MMP-2 and MMP-9. In vivo studies showed that plumbagin inhibited the metastasis of breast cancer cells and decreased osteolytic bone metastases, as well as the secretion of MMP-2 and MMP-9 by tumor cells at metastatic lesions. Conclusions:Plumbagin can suppress the invasion and migration of breast cancer cells via the inhibition of STAT3 signaling and by downregulation of IL-1α, TGF-β, MMP-2 and MMP-9.

Objective:To study the effect of preoperative compound rhizomacurcumaepowder treatment on serum markers and related signaling pathways function in endometrial tissue of patients with endometriosis.Methods:Endometriosis patients in our hospital were chosen for study and randomly divided into the treatment group (n=35) and the control group (n=35). The treatment group received preoperative compound rhizomacurcumaepowder treatment; the control group received preoperative conventional treatment. Then serum markers and function of related signaling pathways in endometrial tissues were compared.Results:After drug treatment, serum SCF, SDC-1, HLA-G, VEGF and Ang-2 contents of treatment group showed a significantly decreasing trend; mRNA contents and protein contents of Wnt, Frizzled, Dsh,β-catenin, cyclinD1, integrinα6β1, ERK1, ERK2, MMP9 and MMP27 in ectopic endometrial tissue of the treatment group all showed a significantly decreasing trend.Conclusion:Preoperative compound rhizomacurcumaepowder treatment is helpful to control the condition of endometriosis and inhibit the activation of cell cycle related signaling pathways and invasion related signaling pathways in ectopic endometrial tissue.