Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Chromones ; pharmacology ; therapeutic use ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia ; metabolism ; pathology ; therapy ; Leupeptins ; pharmacology ; therapeutic use ; Morpholines ; pharmacology ; therapeutic use ; NF-kappa B ; metabolism ; Neoplastic Stem Cells ; drug effects ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors

The purpose of this study was to develop a quantitative and objective method for evaluating neurological deficits in mice with focal cerebral ischemia. After middle cerebral artery occlusion (MCAO), the neurological deficits were evaluated 24 h later. We measured the mean angles, dominant angles and turns in a hanged test in which the mice were sticked on the wall, and the holding angles in an inclined plane test as well, Then we determined the cerebral infarct volumes, neuron density in hippocampus, cortex and subcortical areas 24 h after MCAO. The correlations among infarct volume, neuron density and neurological deficits were analyzed. We also compared the quantitative method with two typical complex methods of behavioral assessment. The effect of [pranlukast, 4-oxo-8-[p-(4-phenylbutyloxy) benzoylamino]-2-(tetrazol-5-yl)-4H-1-benzopyran hemihydrate] (ONO-1078), a neuroprotective agent, on ischemic injury was observed using this method. We found that the variables measured by both quantitative and typical behavioral methods significantly changed in the ischemic mice, and correlated with the infarct volumes and neuron densities. The quantitative variables well correlated with those of typical behavioral assessment, too. ONO-1078 inhibited ischemic injury and reduced the total scores of quantitative assessment. Thus, the quantitative method we developed is useful in evaluating neurological deficits of focal cerebral ischemia with the advantages of objectivity, quantification, simplicity and non-invasion, and can be used in the evaluation of neuroprotective effects of drugs.
Animals ; Behavior, Animal ; Brain ; pathology ; physiopathology ; Brain Ischemia ; etiology ; pathology ; physiopathology ; Chromones ; pharmacology ; therapeutic use ; Female ; Hippocampus ; pathology ; Infarction, Middle Cerebral Artery ; complications ; pathology ; physiopathology ; Leukotriene Antagonists ; pharmacology ; therapeutic use ; Male ; Mice ; Mice, Inbred ICR ; Neurologic Examination ; Neuroprotective Agents ; pharmacology ; therapeutic use

Cancer stem cells (CSCs) have tumor initiation, self-renewal, metastasis and chemo-resistance properties in various tumors including colorectal cancer. Targeting of CSCs may be essential to prevent relapse of tumors after chemotherapy. Phosphatidylinositol-3-kinase (PI3K) and mammalian target of rapamycin (mTOR) signals are central regulators of cell growth, proliferation, differentiation, and apoptosis. These pathways are related to colorectal tumorigenesis. This study focused on PI3K and mTOR pathways by inhibition which initiate differentiation of SW620 derived CSCs and investigated its effect on tumor progression. By using rapamycin, LY294002, and NVP-BEZ235, respectively, PI3K and mTOR signals were blocked independently or dually in colorectal CSCs. Colorectal CSCs gained their differentiation property and lost their stemness properties most significantly in dual-blocked CSCs. After treated with anti-cancer drug (paclitaxel) on the differentiated CSCs cell viability, self-renewal ability and differentiation status were analyzed. As a result dual-blocking group has most enhanced sensitivity for anti-cancer drug. Xenograft tumorigenesis assay by using immunodeficiency mice also shows that dual-inhibited group more effectively increased drug sensitivity and suppressed tumor growth compared to single-inhibited groups. Therefore it could have potent anti-cancer effects that dual-blocking of PI3K and mTOR induces differentiation and improves chemotherapeutic effects on SW620 human colorectal CSCs.
AC133 Antigen/genetics/metabolism ; Animals ; Antineoplastic Agents/pharmacology/therapeutic use ; Cell Differentiation/*drug effects ; Cell Line, Tumor ; Cell Survival/drug effects ; Chromones/pharmacology/therapeutic use ; Colorectal Neoplasms/drug therapy/metabolism/pathology ; Humans ; Imidazoles/pharmacology/therapeutic use ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Morpholines/pharmacology/therapeutic use ; Neoplastic Stem Cells/cytology/drug effects/metabolism ; Paclitaxel/pharmacology/therapeutic use ; Phosphatidylinositol 3-Kinases/*antagonists & inhibitors/metabolism ; Quinolines/pharmacology/therapeutic use ; SOXB1 Transcription Factors/genetics/metabolism ; Signal Transduction/*drug effects ; Sirolimus/pharmacology/therapeutic use ; TOR Serine-Threonine Kinases/*antagonists & inhibitors/metabolism ; Xenograft Model Antitumor Assays