ABSTRACT
OBJECTIVE To investigate the effects of aucubin （Auc） on the malignant biological behavior of breast cancer cells by regulating cyclin-dependent kinase 1（CDK1）/cyclin B1（CCNB1）/Polo-like kinase 1 （PLK1） signaling pathway. METHODS Human breast cancer cells MCF-7 were divided into control group， Auc low-， medium- and high-concentration groups （AUC-L， AUC-M， AUC-H groups， 20， 40 and 80 μmol/L Auc）， Auc-H+pcDNA-NC group （80 μmol/L Auc+transfected pcDNA- NC plasmid）， and Auc-H+pcDNA-CDK1 group （80 μmol/L Auc+transfected pcDNA-CDK1 plasmid）. Cell proliferation， clonal formation， invasion and migration abilities， apoptosis and cycle distribution， and the expressions of related proteins of apoptosis， epithelial-mesenchymal transformation （EMT） and CDK1/CCNB1/PLK1 signaling pathway were detected in each group. The transplanted tumor model of BALB/c nude mice was established by subcutaneous inoculation of MCF-7 cell suspension， and the mice were divided into control group and Auc group （12 mice in each group）. The tumor volume， mass and the expressions of related proteins of CDK1/CCNB1/PLK1 signaling pathway in tumor tissues were detected. RESULTS Compared with control group， the number of clonal formation， proliferation rate， cell invasion number， scar healing rate， G1/G0 phase and S phase cell proportions， and the expressions of B cell lymphoma-2 （Bcl-2）， N-cadherin， fibronectin， CDK1， CCNB1 and PLK1 were decreased significantly （P＜0.05）. The apoptotic rate， G2/M phase cell proportion and the expressions of Bcl-2 associated X protein and E-cadherin were significantly increased， in a dose-dependent manner （P＜0.05）. Compared with the Auc-H+pcDNA-NC group， there was no statistical significance in the above indexes in the Auc-H group （P＞0.05）， while the above indexes in the Auc-H+ pcDNA-CDK1 group were significantly reversed （P＜0.05）. Compared with the control group， the tumor volume and mass， and the expressions of CDK1， CCNB1 and PLK1 in tumor tissue of Auc group were significantly decreased （P＜0.05）. CONCLUSIONS Auc can inhibit the proliferation， invasion and migration of breast cancer cells， induce cell cycle arrest， and inhibit the progression of EMT， which may be related to inhibiting the activation of the CDK1/CCNB1/PLK1 signaling pathway.
ABSTRACT
Objective To detect changes in expression of repressor element silencing transcription factor(REST) in rapid amygdala kindling epilepsy rats. Methods Adult Sprague-Dawley rats were randomly divided into a sham group (group S),epilepsy 2 h group(group EP-2 h),epilepsy 14 d group(group EP-14 d),and epilepsy 35 d group(group EP-35 d). An experimental epilepsy kindling model of the amygdala was established by electrical stimulation. After 2 h, 14 d,and 35 d,rats were perfused and their brains were fixed and frozen for coronal sections. Immunohistochemistry was used to detect expression of REST and the neuron specific nuclear protein, NeuN. Results Expression of REST was significantly increased in the CA1, CA3, and dentate gyrus regions in groups EP compared with group S(P < 0.05), particularly in group EP-2 h. In contrast,NeuN staining was significantly decreased in groups EP compared with group S. Visible neuronal loss was observed in the hippocampus of the EP-35 d group. Conclusions Up-regulation of REST in rat hippocampus may be involved in epileptogenesis.
ABSTRACT
Objective To explore the effect of lentiviral vector-mediated somatostatin (SST) expression on seizures induced by hippocampal kindling in rats.Methods Adult Sprague-Dawley rats were randomly divided into the sham group (Sham),epilepsy group (EP),Lenti-pSyn-EGFP (LV-EGFP) group and Lenti-pSyn-SST-2A-EGFP (LV-SST) group.The rats in LV-EGFP group were subsequently electrically hippocampal kindled and LV-EGFP (5 μl) was injected into dentate gyrus (DG).The rats in LV-SST group were kindled and LV-SST (5 μl) was injected into the dentate gyrus (DG),medial entorhinal cortex (MEC) or amygdala (Amy).Seizure severity was evaluated and immunohistochemical staining was employed to detect the expression of SST,neuron specific nuclear protein (NeuN) and microtubule associated protein 2 (MAP2).Results The current values to the first stage V seizure of LV-SST (DG) group,LV-SST (MEC) group or LV-SST (Amy) group ((143.8±3.8)μA,(142.5±4.1)μA,(142.5±5.3) μA,respectively) were significantly increased compared with that of epilepsy (EP) group ((136.3±5.3)μA),and V stage current values of LV-SST groups in each stimulation day were higher than that of EP group except the fifth stimulation day (P<0.05).After kindling,SST expression and NeuN-positive neurons of EP group and LV-SST groups were less than that of Sham group in CA1,CA3 and DG.SST and NeuN neurons loss in LV-SST groups were less than that of EP group (P<0.05) and MAP2 immunohistochemistry stainings in LV-SST groups were higher than that in EP group.Conclusion Lentiviral vector-mediated somatostatin expression suppresses seizures and can rescue the neuronal damage of seizure induced by kindling in hippocampus,which may provide a new method of gene therapy for temporal lobe epilepsy.