ABSTRACT
OBJECTIVE: Glioma is one of the most common central nervous system malignant tumors, accounting for 45%-60% of adult intracranial tumors. However, the clinical treatment of glioma is limited. It is of great significance to seek new therapeutic methods for glioma via gene therapy. MATERIALS AND METHODS: Microarray is used to identify the lncRNAs that are differentially expressed in glioma. The expression of long non-coding RNA (lncRNA) ROR1-AS1 and miR-4686 was detected by qRT-PCR. Exosomes were isolated from the supernatant of normal and cancerous cells, and TEM was used for exosomes identification. MTT assay, wound healing assay, transwell assay, and colony formation assay were used to detect the exo-ROR1-AS1 function on proliferation, migration, and invasion in glioma cells. Luciferase assay and RIP assay were used to identify the relationship between lncRNA ROR1-AS1 and miR-4686. The effect of exo-ROR1-AS1 on tumorigenesis of glioma was confirmed by the xenograft nude mice model. RESULTS: ROR1-AS1 was up-regulated in glioma tissues, and the high expression of ROR1-AS1 indicated a poor prognosis in glioma patients. Interestingly, ROR1-AS1 was packaged into exosomes and derived from tumor cells. Functional analysis showed exo-ROR1-AS1 promoted the progression of glioma cell lines SHG44 and U251. Furthermore, ROR1-AS1 acted as a sponge of miR-4686 and inhibited its expression. Functionally, forced expression of miR-4686 removed the promoted effects of lncRNA ROR1-AS1 on glioma development. In vivo tumorigenesis experiments showed that exo-ROR1-AS1 promoted glioma development via miR-4686 axis. CONCLUSION: Our study suggested tumor cells derived exo-ROR1-AS1 promoted glioma progression by inhibiting miR-4686, which might be a potential therapeutic target for glioma clinical treatment.
Subject(s)
Disease Progression , Glioma/pathology , RNA, Long Noncoding/genetics , Adult , Animals , Brain Neoplasms/genetics , Brain Neoplasms/pathology , Carcinogenesis , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Cell Transformation, Neoplastic , Glioma/genetics , Humans , Mice , Mice, Nude , Up-Regulation/geneticsABSTRACT
OBJECTIVE: To investigate the dynamic changes of a-AR, b1-AR and b2-AR expression in hepatic fibrosis. METHODS: Rat hepatic fibrosis model was established by bile duct ligation (BDL). HE and Masson staining were used to determine hepatic fibrosis levels. Immunohistochemistry was applied to detect alpha -smooth muscle actin (alpha -SMA), a marker of hepatic stellate cell (HSC) activation; Western blot and real-time RT-PCR were used to measure the dynamic changes of alpha -AR, beta(1)-AR, beta(2)-AR expression on protein and mRNA levels, respectively, during the development of hepatic fibrosis. RESULTS: (1) HE and Masson trichrome staining showed that the liver fibrosis models were established successfully. (2) At 1, 2, 3, 4 wk after BDL, alpha -SMA positive area density of the model group (10.58% +/- 1.75%, 24.14% +/- 2.02%, 29.74% +/- 2.59%, 34.28% +/- 2.01%) was significantly higher than that of the sham operation group (4.12% +/- 1.51%), P less than 0.01. (3) The expression of alpha -AR, beta(1)-AR, beta(2)-AR protein and mRNA was increased with the development of the hepatic fibrosis (P less than 0.05). (4) alpha -SMA expression was positively associated with alpha -AR, beta(1)-AR, beta(2)-AR, r values were 0.564, 0.753 and 0.606, respectively. CONCLUSION: The expression of alpha -SMA is increased dramatically during the fibrosis, and is positively associated with the expression of alpha -AR, beta(1)-AR and beta(2)-AR.
Subject(s)
Actins/metabolism , Liver Cirrhosis, Biliary/metabolism , Liver Cirrhosis, Experimental/metabolism , Receptors, Adrenergic, alpha/metabolism , Receptors, Adrenergic, beta/metabolism , Animals , Hepatic Stellate Cells/metabolism , Hepatic Stellate Cells/pathology , Immunohistochemistry , Liver/metabolism , Liver/pathology , Liver Cirrhosis, Biliary/pathology , Liver Cirrhosis, Experimental/pathology , Male , Polymerase Chain Reaction , RNA, Messenger/genetics , RNA, Messenger/metabolism , Random Allocation , Rats , Rats, Sprague-Dawley , Receptors, Adrenergic, alpha/genetics , Receptors, Adrenergic, beta/genetics , Sympathetic Nervous System/metabolism , Time FactorsABSTRACT
OBJECTIVE: To examine the effects of gamma knife surgery (GKS) on the expression of N-methel-D-asparate receptor (NMDAR) subunits in rat forebrain. MATERIALS AND METHODS: Using stereotactic technique, we performed gamma knife irradiation on the left forebrain of 13 male Wistar rats with a maximum dose of 60 Gy. These animals were raised for 24h, 30 and 60 days before they were killed. Then immunohistochemistry was applied to detect the relative levels of NMDAR subunits (NR1, NR2A, and NR2B) in the target region. RESULTS: The expression of NR1 and NR2A but not NR2B increased significantly in the cortex 30 and 60 days after irradiation. However, no significant differences in the expression of these three subunits were detected in the caudate putamen at all time points. CONCLUSION: gamma knife irradiation induced the upregulation of NMDAR subunits, NR1, and NR2A, which might represent a possible mechanism underlying the therapeutic effects of gamma knife irradiation on many neurological diseases, including drug resistance epilepsy.