Background:Restenosis frequently occurs after percutaneous angioplasty in patients with vascular occlusion and seriously threatens their health.Substantial evidence has revealed that preventing vascular smooth muscle ...Background:Restenosis frequently occurs after percutaneous angioplasty in patients with vascular occlusion and seriously threatens their health.Substantial evidence has revealed that preventing vascular smooth muscle cell proliferation using a drug-eluting stent is an effective approach to improve restenosis.Cucurbitacins have been demonstrated to exert an anti-proliferation effect in various tumors and a hypoten-sive effect.This study aims to investigate the role of cucurbitacins extracted from Cucumis melo L.(CuECs)and cucurbitacin B(CuB)on restenosis.Methods:C57BL/6 mice were subjected to left carotid artery ligation and subcu-taneously injected with CuECs or CuB for 4 weeks.Hematoxylin-Eosin,immuno-fluorescence and immunohistochemistry staining were used to evaluate the effect of CuECs and CuB on neointimal hyperplasia.Western blot,real-time PCR,flow cytometry analysis,EdU staining and cellular immunofluorescence assay were em-ployed to measure the effects of CuECs and CuB on cell proliferation and the cell cycle in vitro.The potential interactions of CuECs with cyclin A2 were performed by molecular docking.Results:The results demonstrated that both CuECs and CuB exhibited significant inhibitory effects on neointimal hyperplasia and proliferation of vascular smooth muscle cells.Furthermore,CuECs and CuB mediated cell cycle arrest at the S phase.Autodocking analysis demonstrated that CuB,CuD,CuE and CuI had high binding en-ergy for cyclin A2.Our study also showed that CuECs and CuB dramatically inhibited FBS-induced cyclin A2 expression.Moreover,the expression of cyclin A2 in CuEC-and CuB-treated neointima was downregulated.Conclusions:CuECs,especially CuB,exert an anti-proliferation effect in VSMCs and may be potential drugs to prevent restenosis.展开更多
Increasing evidence suggests that Cyclin A-Cdk2 activity is required in the apoptosis process induced by various stimuli.To determine a specific substrate of Cyclin A-Cdk2 for apoptosis,in this study,we carried out an...Increasing evidence suggests that Cyclin A-Cdk2 activity is required in the apoptosis process induced by various stimuli.To determine a specific substrate of Cyclin A-Cdk2 for apoptosis,in this study,we carried out an in vitro kinase assay using immunoprecipitated complex Cyclin A-Cdk2 as an enzyme source,and recombinant protein GST-Bad as a substrate.Our study showed that Bad was clearly phosphorylated by Cyclin A-Cdk2 in vitro.To examine whether protein Bad can also be phosphorylated by Cyclin A-Cdk2 kinase in vivo,we transiently overexpressed protein Bad with Cyclin A or Cdk2-dn,a dominant negative version of Cdk2,in Hela cells and determined the phosphorylation status of protein Bad.The test showed that protein Bad was clearly phosphorylated in Cyclin A overexpressed cells,but not in Cdk2-dn or mock transfectent.Moreover,etoposide also caused the phosphorylation of endogenetic Bad.In conclusion,here we provide first time evidence that protein Bad can be a substrate of Cyclin A-Cdk2 apoptosis for in vitro and in vivo.展开更多
In order to investigate the relationship between the expression of cyclin A and drug resistance in adult patients with acute leukemia (AL), the mRNA expression of cyclin A, mdr1, TopⅡ α , bcl-2 was detected in 6...In order to investigate the relationship between the expression of cyclin A and drug resistance in adult patients with acute leukemia (AL), the mRNA expression of cyclin A, mdr1, TopⅡ α , bcl-2 was detected in 64 adult patients with AL and 20 normal controls by semi-reverse transcription polymerse chain reaction (semi-RT-PCR). It was found that the cyclin A and TopⅡ α mRNA expression levels in drug resistant group were significantly lower than in sensitive group ( P <0.01). Under the same experimental condition no cyclin A mRNA expression was detectable in all normal controls. The mdr1 and bcl-2 mRNA expression levels in resistant group were significantly higher than in sensitive group ( P <0.01). cyclin A and TopⅡ α gene expression levels were closely correlated ( r s =+0.794, P=0.000, n =64) in all AL patients, but cyclin A was not correlated with mdr1 and bcl-2 gene expression levels. In drug resistant group there was a negative correlation between the gene expression levels of cyclin A and mdr1 ( r s =-0.337, P=0.029 ). The 10 AL patients with positive lower expression of both cyclin A and TopⅡ α were all resistant to drugs. Logistic regression of Binary analysis showed the correlation between the lower expression of cyclin A and drug resistance. It was concluded that lower expression of cyclin A gene might be an unfavorable prognostic factor for patients with AL, and detection of both cyclin A and TopⅡ α gene expression would predict drug resistance in AL patients.展开更多
Objective To study the relation of the mRNA and protein expression of CyclinA and p21cip1 in different stages hypertrophic scar fibroblast (FB) with its cell cycle,so as to provide theoretical evidence for interventio...Objective To study the relation of the mRNA and protein expression of CyclinA and p21cip1 in different stages hypertrophic scar fibroblast (FB) with its cell cycle,so as to provide theoretical evidence for intervention therapy of展开更多
AIM： To explore the correlation between the expressions of gastrin （GAS）, somatostatin （SS） and cyclin, cyclin- dependent kinase （CDK） in colorectal cancer, and to detect the specific regulatory sites where gas...AIM： To explore the correlation between the expressions of gastrin （GAS）, somatostatin （SS） and cyclin, cyclin- dependent kinase （CDK） in colorectal cancer, and to detect the specific regulatory sites where gastrointestinal hormone regulates cell proliferati6n. METHODS： Seventy-nine resected large intestine carcinomatous specimens were randomly selected. Immunohistochemical staining for GAS, SS, cyclin D1, cyclin E, cyclin A, cyclin B1, CDK2 and CDK4 was performed according to the standard streptavidinbiotin-peroxidase （S-P） method. According to the semiquantitative integral evaluation, SS and GAS were divided into high, middle and low groups. Cyclin D1, cyclin E, cyclin A, cydin B1, CDK2, CDK4 expressions in the three GAS and SS groups were assessed. RESULTS： The positive expression rate of cyclin D1 was significantly higher in high （78.6%, 11/14） and middle GAS groups （73.9%, 17/23） than in low GAS group （45.2%, 19/42） （P〈0.05, X^2 high vs low = 4.691; P〈0.05, X^2 middle vs low = 4.945）. The positive expression rate of cyclin A was significantly higher in high （100%, 14/14） and middle GAS groups （82.6%, 19/23） than in low GAS group （54.8%, 23/42） （P〈0.01, X2high vs low = 9.586; P〈0.05, X^2 middle vs low = 5.040）. The positive expression rate of CDK2 was significantly higher in high （92.9%, 13/14） and middle GAS groups （87.0%, 20/23） than in low GAS group （50.0%, 21142） （P〈0.01, X^2 high vs low = 8.086; P〈0.01,X^2 middle low = 8.715）. The positive expression rate of CDK4 was significantly higher in high （78.6%, 11/14）and middle GAS groups （78.3%, 18/23） than in low GAS group （42.9%, 18/42） （P〈0.05, X^2 high vs low= 5.364; P〈0.01, X^2 middle vs low = 7.539）. The positive expression rate of cyclin E was prominently higher in low SS group （53.3%, 24/45） than in high （9.1%, 1/11） and middle （21.7%, 5/23） SS groups （P〈0.05, X^2 high vs low = 5.325; P〈0.05, X^2 middle vs low = 6.212）. The positive expression rate of CDK2 was significantly higher in low SS group （77.8%, 35/45） than in high SS group （27.3%, 3/11） （P〈0.01, X^2 high vs low = 8.151）. There was a significant positive correlation between the integral ratio of GAS to SS and the semi-quantitative integral of cyclin D1, cyclin E, cyclin A, CDK2, CDK4 （P〈0.05, 0% = 0.252; P〈0.01, E^rs = 0.387; P〈0.01,A^rs = 0.466; P〈0.01, K2^rs = 0.519; P〈0.01, K4^rs = 0.434）. CONCLUSION： The regulation and control of gastrin, SS in colorectal cancer cell growth may be directly related to the abnormal expressions of cyclins D1, A, E, and CDK2, CDK4. The regulatory site of GAS in the cell cycle of colorectal carcinoma may be at the G2, S and G2 phases. The regulatory site of SS may be at the entrance of S phase.展开更多
Cyclin A2 is an essential regulator of the cell division cycle through the activation of kinases that participate to the regulation of S phase as well as the mitotic entry. However,whereas its degradation by the prote...Cyclin A2 is an essential regulator of the cell division cycle through the activation of kinases that participate to the regulation of S phase as well as the mitotic entry. However,whereas its degradation by the proteasome in mid mitosis was thought to be essential for mitosis to proceed,recent observations show that a small fraction of cyclin A2 persists beyond metaphase and is degraded by autophagy. Its implication in the control of cytoskeletal dynamics and cell movement has unveiled its role in the modulation of Rho A activity. Since this GTPase is involved in both cell rounding early in mitosis and later,in the formation of the cleavage furrow,this suggests that cyclin A2 is a novel actor in cytokinesis. Taken together,these data point to this cyclin as a potential mediator of cell-niche interactions whose dysregulation could be taken as a hallmark of metastasis.展开更多
BACKGROUND Cell division cyclin 25C(CDC25C)is a protein that plays a critical role in the cell cycle,specifically in the transition from the G2 phase to the M phase.Recent research has shown that CDC25C could be a pot...BACKGROUND Cell division cyclin 25C(CDC25C)is a protein that plays a critical role in the cell cycle,specifically in the transition from the G2 phase to the M phase.Recent research has shown that CDC25C could be a potential therapeutic target for cancers,particularly for hepatocellular carcinoma(HCC).However,the specific regulatory mechanisms underlying the role of CDC25C in HCC tumorigenesis and development remain incompletely understood.AIM To explore the impact of CDC25C on cell proliferation and apoptosis,as well as its regulatory mechanisms in HCC development.METHODS Hepa1-6 and B16 cells were transduced with a lentiviral vector containing shRNA interference sequences(LV-CDC25C shRNA)to knock down CDC25C.Subsequently,a xenograft mouse model was established by subcutaneously injecting transduced Hepa1-6 cells into C57BL/6 mice to assess the effects of CDC25C knockdown on HCC development in vivo.Cell proliferation and migration were evaluated using a Cell Counting Kit-8 cell proliferation assays and wound healing assays,respectively.The expression of endoplasmic reticulum(ER)stress-related molecules(glucose-regulated protein 78,X-box binding protein-1,and C/EBP homologous protein)was measured in both cells and subcutaneous xenografts using quantitative real-time PCR(qRT-PCR)and western blotting.Additionally,apoptosis was investigated using flow cytometry,qRT-PCR,and western blotting.RESULTS CDC25C was stably suppressed in Hepa1-6 and B16 cells through LV-CDC25C shRNA transduction.A xenograft model with CDC25C knockdown was successfully established and that downregulation of CDC25C expression significantly inhibited HCC growth in mice.CDC25C knockdown not only inhibited cell proliferation and migration but also significantly increased the ER stress response,ultimately promoting ER stress-induced apoptosis in HCC cells.CONCLUSION The regulatory mechanism of CDC25C in HCC development may involve the activation of ER stress and the ER stress-induced apoptosis signaling pathway.展开更多
基金Scientific Research Fund Project of Liaoning Provincial Department of Education,Grant/Award Number:LJKMZ20221267,LJKZ0840 and LJKZ0847National Natural Science Foundation of China Grants,Grant/Award Number:81900267。
文摘Background:Restenosis frequently occurs after percutaneous angioplasty in patients with vascular occlusion and seriously threatens their health.Substantial evidence has revealed that preventing vascular smooth muscle cell proliferation using a drug-eluting stent is an effective approach to improve restenosis.Cucurbitacins have been demonstrated to exert an anti-proliferation effect in various tumors and a hypoten-sive effect.This study aims to investigate the role of cucurbitacins extracted from Cucumis melo L.(CuECs)and cucurbitacin B(CuB)on restenosis.Methods:C57BL/6 mice were subjected to left carotid artery ligation and subcu-taneously injected with CuECs or CuB for 4 weeks.Hematoxylin-Eosin,immuno-fluorescence and immunohistochemistry staining were used to evaluate the effect of CuECs and CuB on neointimal hyperplasia.Western blot,real-time PCR,flow cytometry analysis,EdU staining and cellular immunofluorescence assay were em-ployed to measure the effects of CuECs and CuB on cell proliferation and the cell cycle in vitro.The potential interactions of CuECs with cyclin A2 were performed by molecular docking.Results:The results demonstrated that both CuECs and CuB exhibited significant inhibitory effects on neointimal hyperplasia and proliferation of vascular smooth muscle cells.Furthermore,CuECs and CuB mediated cell cycle arrest at the S phase.Autodocking analysis demonstrated that CuB,CuD,CuE and CuI had high binding en-ergy for cyclin A2.Our study also showed that CuECs and CuB dramatically inhibited FBS-induced cyclin A2 expression.Moreover,the expression of cyclin A2 in CuEC-and CuB-treated neointima was downregulated.Conclusions:CuECs,especially CuB,exert an anti-proliferation effect in VSMCs and may be potential drugs to prevent restenosis.
基金Supported by the National Natural Science Foundation of China(Nos.30370294and30640064).
文摘Increasing evidence suggests that Cyclin A-Cdk2 activity is required in the apoptosis process induced by various stimuli.To determine a specific substrate of Cyclin A-Cdk2 for apoptosis,in this study,we carried out an in vitro kinase assay using immunoprecipitated complex Cyclin A-Cdk2 as an enzyme source,and recombinant protein GST-Bad as a substrate.Our study showed that Bad was clearly phosphorylated by Cyclin A-Cdk2 in vitro.To examine whether protein Bad can also be phosphorylated by Cyclin A-Cdk2 kinase in vivo,we transiently overexpressed protein Bad with Cyclin A or Cdk2-dn,a dominant negative version of Cdk2,in Hela cells and determined the phosphorylation status of protein Bad.The test showed that protein Bad was clearly phosphorylated in Cyclin A overexpressed cells,but not in Cdk2-dn or mock transfectent.Moreover,etoposide also caused the phosphorylation of endogenetic Bad.In conclusion,here we provide first time evidence that protein Bad can be a substrate of Cyclin A-Cdk2 apoptosis for in vitro and in vivo.
文摘In order to investigate the relationship between the expression of cyclin A and drug resistance in adult patients with acute leukemia (AL), the mRNA expression of cyclin A, mdr1, TopⅡ α , bcl-2 was detected in 64 adult patients with AL and 20 normal controls by semi-reverse transcription polymerse chain reaction (semi-RT-PCR). It was found that the cyclin A and TopⅡ α mRNA expression levels in drug resistant group were significantly lower than in sensitive group ( P <0.01). Under the same experimental condition no cyclin A mRNA expression was detectable in all normal controls. The mdr1 and bcl-2 mRNA expression levels in resistant group were significantly higher than in sensitive group ( P <0.01). cyclin A and TopⅡ α gene expression levels were closely correlated ( r s =+0.794, P=0.000, n =64) in all AL patients, but cyclin A was not correlated with mdr1 and bcl-2 gene expression levels. In drug resistant group there was a negative correlation between the gene expression levels of cyclin A and mdr1 ( r s =-0.337, P=0.029 ). The 10 AL patients with positive lower expression of both cyclin A and TopⅡ α were all resistant to drugs. Logistic regression of Binary analysis showed the correlation between the lower expression of cyclin A and drug resistance. It was concluded that lower expression of cyclin A gene might be an unfavorable prognostic factor for patients with AL, and detection of both cyclin A and TopⅡ α gene expression would predict drug resistance in AL patients.
文摘Objective To study the relation of the mRNA and protein expression of CyclinA and p21cip1 in different stages hypertrophic scar fibroblast (FB) with its cell cycle,so as to provide theoretical evidence for intervention therapy of
基金Supported by the Natural Science Foundation of Anhui Province,No.03043704
文摘AIM： To explore the correlation between the expressions of gastrin （GAS）, somatostatin （SS） and cyclin, cyclin- dependent kinase （CDK） in colorectal cancer, and to detect the specific regulatory sites where gastrointestinal hormone regulates cell proliferati6n. METHODS： Seventy-nine resected large intestine carcinomatous specimens were randomly selected. Immunohistochemical staining for GAS, SS, cyclin D1, cyclin E, cyclin A, cyclin B1, CDK2 and CDK4 was performed according to the standard streptavidinbiotin-peroxidase （S-P） method. According to the semiquantitative integral evaluation, SS and GAS were divided into high, middle and low groups. Cyclin D1, cyclin E, cyclin A, cydin B1, CDK2, CDK4 expressions in the three GAS and SS groups were assessed. RESULTS： The positive expression rate of cyclin D1 was significantly higher in high （78.6%, 11/14） and middle GAS groups （73.9%, 17/23） than in low GAS group （45.2%, 19/42） （P〈0.05, X^2 high vs low = 4.691; P〈0.05, X^2 middle vs low = 4.945）. The positive expression rate of cyclin A was significantly higher in high （100%, 14/14） and middle GAS groups （82.6%, 19/23） than in low GAS group （54.8%, 23/42） （P〈0.01, X2high vs low = 9.586; P〈0.05, X^2 middle vs low = 5.040）. The positive expression rate of CDK2 was significantly higher in high （92.9%, 13/14） and middle GAS groups （87.0%, 20/23） than in low GAS group （50.0%, 21142） （P〈0.01, X^2 high vs low = 8.086; P〈0.01,X^2 middle low = 8.715）. The positive expression rate of CDK4 was significantly higher in high （78.6%, 11/14）and middle GAS groups （78.3%, 18/23） than in low GAS group （42.9%, 18/42） （P〈0.05, X^2 high vs low= 5.364; P〈0.01, X^2 middle vs low = 7.539）. The positive expression rate of cyclin E was prominently higher in low SS group （53.3%, 24/45） than in high （9.1%, 1/11） and middle （21.7%, 5/23） SS groups （P〈0.05, X^2 high vs low = 5.325; P〈0.05, X^2 middle vs low = 6.212）. The positive expression rate of CDK2 was significantly higher in low SS group （77.8%, 35/45） than in high SS group （27.3%, 3/11） （P〈0.01, X^2 high vs low = 8.151）. There was a significant positive correlation between the integral ratio of GAS to SS and the semi-quantitative integral of cyclin D1, cyclin E, cyclin A, CDK2, CDK4 （P〈0.05, 0% = 0.252; P〈0.01, E^rs = 0.387; P〈0.01,A^rs = 0.466; P〈0.01, K2^rs = 0.519; P〈0.01, K4^rs = 0.434）. CONCLUSION： The regulation and control of gastrin, SS in colorectal cancer cell growth may be directly related to the abnormal expressions of cyclins D1, A, E, and CDK2, CDK4. The regulatory site of GAS in the cell cycle of colorectal carcinoma may be at the G2, S and G2 phases. The regulatory site of SS may be at the entrance of S phase.
基金Supported by Agence Nationale de la Recherche(08-BLAN-0037-02),Fondation ARC pour la Recherche sur le Cancerand Gefluc+2 种基金CNRS/Région Languedoc-Roussillon and Fondation pour la Recherche Médicale(Loukil A)the French Ministry of Education and Research and Fondation pour la Recherche Médicale(Bendris N)the Canadian Institutes of Health Research,La Ligue Contre le Cancer and the Fondation de France(Cheung CT)
文摘Cyclin A2 is an essential regulator of the cell division cycle through the activation of kinases that participate to the regulation of S phase as well as the mitotic entry. However,whereas its degradation by the proteasome in mid mitosis was thought to be essential for mitosis to proceed,recent observations show that a small fraction of cyclin A2 persists beyond metaphase and is degraded by autophagy. Its implication in the control of cytoskeletal dynamics and cell movement has unveiled its role in the modulation of Rho A activity. Since this GTPase is involved in both cell rounding early in mitosis and later,in the formation of the cleavage furrow,this suggests that cyclin A2 is a novel actor in cytokinesis. Taken together,these data point to this cyclin as a potential mediator of cell-niche interactions whose dysregulation could be taken as a hallmark of metastasis.
基金Supported by Natural Science Foundation of Guangxi Zhuang Autonomous Region,China,No.2023GXNSFAA026070 and No.2018GXNSFAA281071.
文摘BACKGROUND Cell division cyclin 25C(CDC25C)is a protein that plays a critical role in the cell cycle,specifically in the transition from the G2 phase to the M phase.Recent research has shown that CDC25C could be a potential therapeutic target for cancers,particularly for hepatocellular carcinoma(HCC).However,the specific regulatory mechanisms underlying the role of CDC25C in HCC tumorigenesis and development remain incompletely understood.AIM To explore the impact of CDC25C on cell proliferation and apoptosis,as well as its regulatory mechanisms in HCC development.METHODS Hepa1-6 and B16 cells were transduced with a lentiviral vector containing shRNA interference sequences(LV-CDC25C shRNA)to knock down CDC25C.Subsequently,a xenograft mouse model was established by subcutaneously injecting transduced Hepa1-6 cells into C57BL/6 mice to assess the effects of CDC25C knockdown on HCC development in vivo.Cell proliferation and migration were evaluated using a Cell Counting Kit-8 cell proliferation assays and wound healing assays,respectively.The expression of endoplasmic reticulum(ER)stress-related molecules(glucose-regulated protein 78,X-box binding protein-1,and C/EBP homologous protein)was measured in both cells and subcutaneous xenografts using quantitative real-time PCR(qRT-PCR)and western blotting.Additionally,apoptosis was investigated using flow cytometry,qRT-PCR,and western blotting.RESULTS CDC25C was stably suppressed in Hepa1-6 and B16 cells through LV-CDC25C shRNA transduction.A xenograft model with CDC25C knockdown was successfully established and that downregulation of CDC25C expression significantly inhibited HCC growth in mice.CDC25C knockdown not only inhibited cell proliferation and migration but also significantly increased the ER stress response,ultimately promoting ER stress-induced apoptosis in HCC cells.CONCLUSION The regulatory mechanism of CDC25C in HCC development may involve the activation of ER stress and the ER stress-induced apoptosis signaling pathway.
