ABSTRACT
OBJECTIVE@#To study the expression of miRNA 320a in the brain tissue of epileptic rats and analyze its effect on the expression of aquaporin 4 (AQP4).@*METHODS@#All rats were performed with the intraperitoneal injection of lithium chloride (3 mmol/kg) and then the intraperitoneal injection of pilocarpine (30 mg/kg) 24 h later (injected twice) to prepare the epileptic model of Wistar rats. Rats in the control group were injected with the equal volume of normal saline. According to the Racine scale, rats with over stage 3 of epilepsy were chosen and the brain tissue was separated quickly and then stored at -80 °C. The immunohistochemistry was used to detect the expression of aquaporin in the brain tissue of epileptic model and the Real-time PCR was employed to determine the difference in the expression of miRNA 320a and AQP4 in the brain tissue of rats between the epileptic model group and control group. Five 5-day neonatal Wistar rats were chosen to collect the cerebral cortex and their primary astrocytes were separated and cultured. They were transfected with miRNA mimic and imitated to the endogenous miRNA 320a to up-regulate the expression of miRNA 320a.@*RESULTS@#In the model group, the expression of AQP4 was significantly higher than the control group (P < 0.01). However, the expression of miRNA 320a in the model group was lower than control group (P < 0.05), which was negatively correlated to AQP4. In the primary astrocytes, the transfection of miRNA 320a mimic could significantly reduce the expression of AQP4, while its inhibitor could up-regulate the expression of AQP4, which indicated that miRNA 320a could reduce the expression of AQP4.@*CONCLUSIONS@#In the primary astrocytes of rats, the miRNA 320a could inhibit the expression of AQP4 and after adding the inhibitor of miRNA 320a, the expression of AQP4 was up-regulated.
ABSTRACT
Objective: To study the expression of miRNA 320a in the brain tissue of epileptic rats and analyze its effect on the expression of aquaporin 4 (AQP4). Methods: All rats were performed with the intraperitoneal injection of lithium chloride (3 mmol/kg) and then the intraperitoneal injection of pilocarpine (30 mg/kg) 24 h later (injected twice) to prepare the epileptic model of Wistar rats. Rats in the control group were injected with the equal volume of normal saline. According to the Racine scale, rats with over stage 3 of epilepsy were chosen and the brain tissue was separated quickly and then stored at -80 °C. The immunohistochemistry was used to detect the expression of aquaporin in the brain tissue of epileptic model and the Real-time PCR was employed to determine the difference in the expression of miRNA 320a and AQP4 in the brain tissue of rats between the epileptic model group and control group. Five 5-day neonatal Wistar rats were chosen to collect the cerebral cortex and their primary astrocytes were separated and cultured. They were transfected with miRNA mimic and imitated to the endogenous miRNA 320a to up-regulate the expression of miRNA 320a. Results: In the model group, the expression of AQP4 was significantly higher than the control group (P < 0.01). However, the expression of miRNA 320a in the model group was lower than control group (P < 0.05), which was negatively correlated to AQP4. In the primary astrocytes, the transfection of miRNA 320a mimic could significantly reduce the expression of AQP4, while its inhibitor could up-regulate the expression of AQP4, which indicated that miRNA 320a could reduce the expression of AQP4. Conclusions: In the primary astrocytes of rats, the miRNA 320a could inhibit the expression of AQP4 and after adding the inhibitor of miRNA 320a, the expression of AQP4 was up-regulated.
ABSTRACT
Background and Aim: The strategies of targeting valosin-containing protein (VCP) may have therapeutic potential for treating cancer metastasis. In this study, we aim to investigate the correlation of VCP protein expression in osteosarcoma (OS) tissues with pulmonary metastasis and its possible molecular mechanism. Materials and Methods: Expression of VCP in 60 OS specimens was detected by immunohistochemistry (IHC) and the relationship with metastasis was analyzed. An artifi cial micro ribonucleic acid, targeting VCP, was performed to silence the expression of VCP in U2-OS cells. Cell mobility was detected by wound healing and Transwell assays. Western blot and real-time polymerase chain reaction were performed to investigate the expression of VCP in U2-OS cells. Furthermore, the protein of pAKT (phosphorylated serine/threonine protein kinase) and nuclear factor of kappa B protein65 were measured by western blot to evaluate the effect of silencing VCP on AKT/nuclear factor of kappa B (NF-B) signaling pathway. Results: The results showed that the expression level of VCP protein in cases with pulmonary metastases was signifi cantly higher than that in those without metastasis (P = 0.004). The invasion and migration of U2-OS cells were suppressed by silencing VCP. Furthermore, silencing VCP could down-regulate the phosphorylation of AKT and nuclear transfer of NF-B. Conclusions: Our fi ndings suggested that inhibition of VCP could suppress OS cells invasion and migration through down-regulating AKT/NF-B signaling pathway.
ABSTRACT
Background : Fatty acid synthase (FASN) is overexpressed in a variety of human cancers, and may be involved in cancer metastasis. Hence, the strategies targeted on FASN may have therapeutic potential for treating cancer metastasis. Objectives : The aim of this study is to investigate the correlation of FASN expression with metastasis in human osteosarcoma. Materials and Methods : Human osteosarcoma cell lines U2-OS and osteosarcoma biopsy specimens were employed in this study. The expression of FASN protein in osteosarcoma specimens was detected by IHC (immunohistochemistry) and the relationship with metastasis was analyzed. We performed the cerulenin, an inhibitor of FASN, to inhibit FASN expression in U2-OS cells. Western blot and RT-PCR were performed to investigate the expression of FASN in U2-OS cells. Cells mobility was detected by wound healing and Transwell assays. Results : Results showed that the FASN expression level in the cases with pulmonary metastases was significantly higher than in those without metastasis. In vitro, the invasion and migration of U2-OS cells were suppressed by inhibiting FASN. Our findings suggested that FASN may be involved in osteosarcoma metastasis