Purpose:To study the effects of high-dose （）60Co gamma ray to the cell cycles and cell cyclin D1,B1 and cyclin dependent kinase（CDK）1,4 of nasopharygeal carcinoma cell line CNE-2. Observe consecutively the mechani...Purpose:To study the effects of high-dose （）60Co gamma ray to the cell cycles and cell cyclin D1,B1 and cyclin dependent kinase（CDK）1,4 of nasopharygeal carcinoma cell line CNE-2. Observe consecutively the mechanism of the action of high-dose gamma ray on cell cycle progress, in order So as to provide basic material for hypofractionated radiotherapy schedule for tumors.Methods:Propidium Iodide（PI） simple staining and flow cytometry（FC） were used to detect the cell cycle. The expression of cyclin D1、cyclinB1、CDK4 and CDK1 mRNA were detected by combining RT-PCR with semi-quantitative analysis.Results:Apparent G2M phase arrest was observed first after CNE-2 cells were exposed to 4.0～8.0Gy fractionated radiation with gamma rays. G1 phase arrest was enhanced with increasing absorbed dose. Then all cell phase were arrested, but the dose-dependent relation was not obvious. The expression of cyclin D1、cyclin B1 mRNA was suppressed after CNE-2 cells were exposed to 4.0～8.0Gy gamma rays（P<0.05）. There was no significant difference between the expression of CDK4 mRNA and the control group（P>0.05）. But the CDK1 mRNA was not expressed in any of the irradiated groups.Conclusions:G1 phase, S phase and G2M phase arrest were observed in CNE-2 cells exposed to 4.0～8.0Gy fractionated radiation in gamma rays. But the dose-dependent relation was not obvious. The main effects were that the cyclin D1 activity suppression lead to G1 phase arrest ,and G2M arrest was correlated to the cyclin B1/CDK1 activity after 4.0～8.0 Gy gamma rays.[展开更多
Mot-2 protein is shown to interact with p53 and inhibit its transcriptional activation function. Mot-2 overexpressing stable clones of NIH 3T3 cells were malignantly transformed, however, they had a high level of expr...Mot-2 protein is shown to interact with p53 and inhibit its transcriptional activation function. Mot-2 overexpressing stable clones of NIH 3T3 cells were malignantly transformed, however, they had a high level of expression of a p53 downstream gene, p21WAF1. The present study was undertaken to elucidate possible molecular mechanism(s) of such upregulation. An inCreased level of p21WAF1, expression was detected in sta- ble transfectants although an exogenous reporter gene driven by p21WAF1, promoter exhibited lower activity in these cells suggesting that some post-transcriptional mechanism contributes to upregulation. Western analyses of transient and stable clones revealed that upregulation of p21WAF1, in stable NIH 3T3/mot-2 cells may be mediated by cyclin D1 and cdk-2.展开更多
CDK<span style="white-space:nowrap;"><sub><sub><span style="font-family:Verdana;">2<span style="white-space:nowrap;"></sub></sub><span style=&...CDK<span style="white-space:nowrap;"><sub><sub><span style="font-family:Verdana;">2<span style="white-space:nowrap;"></sub></sub><span style="font-family:Verdana;"> is one of the most important members of Cyclin-dependent kinases. It is a critical modulator of various oncogenic signaling pathways, and its activity is vital for <span style="font-family:Verdana;">loss<span style="font-family:Verdana;"> of proliferative control during oncogenesis. This work has focused on developing a pharmacophore model for CDK<span style="white-space:nowrap;"><sub><sub><span style="font-family:Verdana;">2<span style="white-space:nowrap;"></sub></sub><span style="font-family:Verdana;"> inhibitors by using a dataset of known inhibitors as a pre-filter throughout the virtual screening and docking process. Consequently, the best pharmacophore model was made of one hydrogen bond acceptor, and two aromatic ring features with <span style="font-family:Verdana;">a <span style="font-family:Verdana;">high<span style="font-family:""><span style="font-family:Verdana;"> correlation value of 0.906. The validation findings proved out that the selected model can be used as a filter to screen new molecules like Enamine kinase hinge region directed library against CDK<span style="white-space:nowrap;"><sub><sub><span style="font-family:Verdana;">2<span style="white-space:nowrap;"></sub></sub><span style="font-family:Verdana;">. As a result, 69 hits were subjected to molecular docking studies. Eventually, three compounds<span style="font-family:Verdana;"> (<span style="font-family:""><span style="font-family:Verdana;">5909, 701 <span style="font-family:Verdana;">and<span style="font-family:Verdana;"> 8397<span style="font-family:Verdana;">) <span style="font-family:""><span style="font-family:Verdana;">scored good interaction energy values and strong molecular interactions. Hence, they were identified as leads for novel CDK<span style="white-space:nowrap;"><sub><sub><span style="font-family:Verdana;">2<span style="white-space:nowrap;"></sub></sub><span style="font-family:Verdana;"> inhibitors as anticancer drugs.展开更多
文摘Purpose:To study the effects of high-dose （）60Co gamma ray to the cell cycles and cell cyclin D1,B1 and cyclin dependent kinase（CDK）1,4 of nasopharygeal carcinoma cell line CNE-2. Observe consecutively the mechanism of the action of high-dose gamma ray on cell cycle progress, in order So as to provide basic material for hypofractionated radiotherapy schedule for tumors.Methods:Propidium Iodide（PI） simple staining and flow cytometry（FC） were used to detect the cell cycle. The expression of cyclin D1、cyclinB1、CDK4 and CDK1 mRNA were detected by combining RT-PCR with semi-quantitative analysis.Results:Apparent G2M phase arrest was observed first after CNE-2 cells were exposed to 4.0～8.0Gy fractionated radiation with gamma rays. G1 phase arrest was enhanced with increasing absorbed dose. Then all cell phase were arrested, but the dose-dependent relation was not obvious. The expression of cyclin D1、cyclin B1 mRNA was suppressed after CNE-2 cells were exposed to 4.0～8.0Gy gamma rays（P<0.05）. There was no significant difference between the expression of CDK4 mRNA and the control group（P>0.05）. But the CDK1 mRNA was not expressed in any of the irradiated groups.Conclusions:G1 phase, S phase and G2M phase arrest were observed in CNE-2 cells exposed to 4.0～8.0Gy fractionated radiation in gamma rays. But the dose-dependent relation was not obvious. The main effects were that the cyclin D1 activity suppression lead to G1 phase arrest ,and G2M arrest was correlated to the cyclin B1/CDK1 activity after 4.0～8.0 Gy gamma rays.[
文摘Mot-2 protein is shown to interact with p53 and inhibit its transcriptional activation function. Mot-2 overexpressing stable clones of NIH 3T3 cells were malignantly transformed, however, they had a high level of expression of a p53 downstream gene, p21WAF1. The present study was undertaken to elucidate possible molecular mechanism(s) of such upregulation. An inCreased level of p21WAF1, expression was detected in sta- ble transfectants although an exogenous reporter gene driven by p21WAF1, promoter exhibited lower activity in these cells suggesting that some post-transcriptional mechanism contributes to upregulation. Western analyses of transient and stable clones revealed that upregulation of p21WAF1, in stable NIH 3T3/mot-2 cells may be mediated by cyclin D1 and cdk-2.
文摘CDK<span style="white-space:nowrap;"><sub><sub><span style="font-family:Verdana;">2<span style="white-space:nowrap;"></sub></sub><span style="font-family:Verdana;"> is one of the most important members of Cyclin-dependent kinases. It is a critical modulator of various oncogenic signaling pathways, and its activity is vital for <span style="font-family:Verdana;">loss<span style="font-family:Verdana;"> of proliferative control during oncogenesis. This work has focused on developing a pharmacophore model for CDK<span style="white-space:nowrap;"><sub><sub><span style="font-family:Verdana;">2<span style="white-space:nowrap;"></sub></sub><span style="font-family:Verdana;"> inhibitors by using a dataset of known inhibitors as a pre-filter throughout the virtual screening and docking process. Consequently, the best pharmacophore model was made of one hydrogen bond acceptor, and two aromatic ring features with <span style="font-family:Verdana;">a <span style="font-family:Verdana;">high<span style="font-family:""><span style="font-family:Verdana;"> correlation value of 0.906. The validation findings proved out that the selected model can be used as a filter to screen new molecules like Enamine kinase hinge region directed library against CDK<span style="white-space:nowrap;"><sub><sub><span style="font-family:Verdana;">2<span style="white-space:nowrap;"></sub></sub><span style="font-family:Verdana;">. As a result, 69 hits were subjected to molecular docking studies. Eventually, three compounds<span style="font-family:Verdana;"> (<span style="font-family:""><span style="font-family:Verdana;">5909, 701 <span style="font-family:Verdana;">and<span style="font-family:Verdana;"> 8397<span style="font-family:Verdana;">) <span style="font-family:""><span style="font-family:Verdana;">scored good interaction energy values and strong molecular interactions. Hence, they were identified as leads for novel CDK<span style="white-space:nowrap;"><sub><sub><span style="font-family:Verdana;">2<span style="white-space:nowrap;"></sub></sub><span style="font-family:Verdana;"> inhibitors as anticancer drugs.
