目的:探讨大豆异黄酮对小鼠肝癌移植瘤细胞凋亡的诱导机制。方法:建立小鼠H22肝癌皮下移植瘤模型,分为模型组、大豆异黄酮组和5-氟尿嘧啶组。采用DNA ladder法检测移植瘤细胞凋亡情况,蛋白印迹法检测移植瘤组织含半胱氨酸的天冬氨酸蛋...目的:探讨大豆异黄酮对小鼠肝癌移植瘤细胞凋亡的诱导机制。方法:建立小鼠H22肝癌皮下移植瘤模型,分为模型组、大豆异黄酮组和5-氟尿嘧啶组。采用DNA ladder法检测移植瘤细胞凋亡情况,蛋白印迹法检测移植瘤组织含半胱氨酸的天冬氨酸蛋白水解酶(cysteinyl aspartate specific proteinase,Caspase)-3、Caspase-8、Caspase-9、B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2-associated X protein,Bax)、细胞色素c(cytochrome c,Cyt c)、细胞凋亡诱导因子(apoptosis inducing factor,AIF)、p53、Survivin和信号传导子及转录激活子(signal transducer and activator of transcription,STAT)3表达情况。结果:与模型组比较,大豆异黄酮组移植瘤质量减轻;肿瘤细胞凋亡特征性DNA片段增多;移植瘤细胞Caspase-3、Caspase-8和Caspase-9的活性片段增多;Bcl-2蛋白相对表达量降低,Bax蛋白相对表达量升高;胞浆Cyt c和AIF水平升高。同时,大豆异黄酮上调移植瘤组织p53蛋白表达,下调STAT3活化和Survivin蛋白表达。结论:大豆异黄酮诱导H22移植瘤细胞凋亡,其促凋亡机制可能与p53和STAT3有关。展开更多
老龄化可引起铁元素在脑内的蓄积。随着年龄的增加,脉络丛上皮可能是脑铁毒性、炎症反应和氧化应激损伤的重要结构。本文旨在研究铁代谢调节中的关键激素——铁调素在脉络丛上皮表达的年龄变化,及其与白细胞介素-6（interleukin-6,IL-6...老龄化可引起铁元素在脑内的蓄积。随着年龄的增加,脉络丛上皮可能是脑铁毒性、炎症反应和氧化应激损伤的重要结构。本文旨在研究铁代谢调节中的关键激素——铁调素在脉络丛上皮表达的年龄变化,及其与白细胞介素-6（interleukin-6,IL-6）和信号转导子及转录激活子3（signal transducer and activator of transcription 3,Stat3）的关系。本文以不同年龄段（3、6、9、12、15、18、21、24、27、30、33和36月龄）大鼠为研究对象,用实时定量RT-PCR和Western blot法检测铁调素及IL-6/Stat3信号途径相关蛋白在脉络丛上皮的表达变化。结果显示,3~18月龄年轻大鼠脉络丛上皮铁调素m RNA没有明显变化,21~36月龄大鼠的铁调素m RNA水平显著增加。脉络丛上皮IL-6和p-Stat3 m RNA和蛋白表达的年龄变化趋势与铁调素m RNA一致。以上结果提示,随着年龄的增加,炎症因子IL-6可能作用于相应的受体,通过Stat3信号转导途径,参与调节铁调素在脉络丛上皮的表达变化。展开更多
The thioredoxin system plays a role in a variety of physiological functions, including cell growth, differenti- ation, apoptosis, tumorigenesis, and immunity. We previously confirmed that butaselen （BS）, a novel thi...The thioredoxin system plays a role in a variety of physiological functions, including cell growth, differenti- ation, apoptosis, tumorigenesis, and immunity. We previously confirmed that butaselen （BS）, a novel thioredoxin reductase inhibitor, can inhibit the growth of various human cancer cell lines, yet the underlying mechanism remains elusive. In this study, we investigated the anti-tumor effect of BS in vivo through regulating the immune system of KM mice. We found that BS inhibits tumor proliferation by promoting the activation of splenic lymphocytes in mice. BS can elevate the percentage of CD^4-CD8^＋ T lymphocytes and the secretion of downstream cytokines in mice via downregulating the expression of programmed death-ligand 1 （PD-L1） on the tumor cells＇ surface in vivo. Further study in HepG2 and BEL-7402 cells showed that decrease of PD-L1 level after BS treatment was achieved by inhibiting signal transducer and activator of transcription 3 （STAT3） phosphorylation. Taken together, our results suggest that BS has a role in promoting the immune response by reducing PD-L1 expression via the STAT3 pathway, and subsequently suppresses tumorigenesis.展开更多
目的构建并鉴定靶向大鼠血管平滑肌细胞信号传导子及转录激活子3(STAT3)基因的慢病毒干扰载体。方法针对STAT3基因的不同部位设计4对短发卡RNA(shRNA)的寡核苷酸片段,克隆到慢病毒干扰载体PLKO.1中,构建shRNA慢病毒干扰载体PLKO.1-STAT3...目的构建并鉴定靶向大鼠血管平滑肌细胞信号传导子及转录激活子3(STAT3)基因的慢病毒干扰载体。方法针对STAT3基因的不同部位设计4对短发卡RNA(shRNA)的寡核苷酸片段,克隆到慢病毒干扰载体PLKO.1中,构建shRNA慢病毒干扰载体PLKO.1-STAT3-shRNA-1、2、3、4,并感染大鼠胸主动脉血管平滑肌细胞。Western blot法检测并筛选最佳抑制效率的shRNA干扰载体。结果 Western blot法证实PLKO.1-STAT3-shRNA-1、2、3、4均能抑制细胞内STAT3基因的表达,以PLKO.1-STAT3-S1最为明显。用PLKO.1-STAT3-S1转染血管平滑肌细胞后可以促进细胞凋亡。结论成功构建了靶向STAT3基因的shRNA慢病毒干扰载体。展开更多
文摘目的:探讨大豆异黄酮对小鼠肝癌移植瘤细胞凋亡的诱导机制。方法:建立小鼠H22肝癌皮下移植瘤模型,分为模型组、大豆异黄酮组和5-氟尿嘧啶组。采用DNA ladder法检测移植瘤细胞凋亡情况,蛋白印迹法检测移植瘤组织含半胱氨酸的天冬氨酸蛋白水解酶(cysteinyl aspartate specific proteinase,Caspase)-3、Caspase-8、Caspase-9、B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2-associated X protein,Bax)、细胞色素c(cytochrome c,Cyt c)、细胞凋亡诱导因子(apoptosis inducing factor,AIF)、p53、Survivin和信号传导子及转录激活子(signal transducer and activator of transcription,STAT)3表达情况。结果:与模型组比较,大豆异黄酮组移植瘤质量减轻;肿瘤细胞凋亡特征性DNA片段增多;移植瘤细胞Caspase-3、Caspase-8和Caspase-9的活性片段增多;Bcl-2蛋白相对表达量降低,Bax蛋白相对表达量升高;胞浆Cyt c和AIF水平升高。同时,大豆异黄酮上调移植瘤组织p53蛋白表达,下调STAT3活化和Survivin蛋白表达。结论:大豆异黄酮诱导H22移植瘤细胞凋亡,其促凋亡机制可能与p53和STAT3有关。
基金supported by the Natural Science Foundation of Zhejiang Province,China(No.Y2110388)the Science and Technology Project of Zhejiang Province,China(No.Y2011C37091)
文摘老龄化可引起铁元素在脑内的蓄积。随着年龄的增加,脉络丛上皮可能是脑铁毒性、炎症反应和氧化应激损伤的重要结构。本文旨在研究铁代谢调节中的关键激素——铁调素在脉络丛上皮表达的年龄变化,及其与白细胞介素-6（interleukin-6,IL-6）和信号转导子及转录激活子3（signal transducer and activator of transcription 3,Stat3）的关系。本文以不同年龄段（3、6、9、12、15、18、21、24、27、30、33和36月龄）大鼠为研究对象,用实时定量RT-PCR和Western blot法检测铁调素及IL-6/Stat3信号途径相关蛋白在脉络丛上皮的表达变化。结果显示,3~18月龄年轻大鼠脉络丛上皮铁调素m RNA没有明显变化,21~36月龄大鼠的铁调素m RNA水平显著增加。脉络丛上皮IL-6和p-Stat3 m RNA和蛋白表达的年龄变化趋势与铁调素m RNA一致。以上结果提示,随着年龄的增加,炎症因子IL-6可能作用于相应的受体,通过Stat3信号转导途径,参与调节铁调素在脉络丛上皮的表达变化。
基金Project supported by the National Natural Science Foundation of China(No.81372266)the National Science and Technology Major Project(No.2011zx09101-001-03)China
文摘The thioredoxin system plays a role in a variety of physiological functions, including cell growth, differenti- ation, apoptosis, tumorigenesis, and immunity. We previously confirmed that butaselen （BS）, a novel thioredoxin reductase inhibitor, can inhibit the growth of various human cancer cell lines, yet the underlying mechanism remains elusive. In this study, we investigated the anti-tumor effect of BS in vivo through regulating the immune system of KM mice. We found that BS inhibits tumor proliferation by promoting the activation of splenic lymphocytes in mice. BS can elevate the percentage of CD^4-CD8^＋ T lymphocytes and the secretion of downstream cytokines in mice via downregulating the expression of programmed death-ligand 1 （PD-L1） on the tumor cells＇ surface in vivo. Further study in HepG2 and BEL-7402 cells showed that decrease of PD-L1 level after BS treatment was achieved by inhibiting signal transducer and activator of transcription 3 （STAT3） phosphorylation. Taken together, our results suggest that BS has a role in promoting the immune response by reducing PD-L1 expression via the STAT3 pathway, and subsequently suppresses tumorigenesis.
文摘目的构建并鉴定靶向大鼠血管平滑肌细胞信号传导子及转录激活子3(STAT3)基因的慢病毒干扰载体。方法针对STAT3基因的不同部位设计4对短发卡RNA(shRNA)的寡核苷酸片段,克隆到慢病毒干扰载体PLKO.1中,构建shRNA慢病毒干扰载体PLKO.1-STAT3-shRNA-1、2、3、4,并感染大鼠胸主动脉血管平滑肌细胞。Western blot法检测并筛选最佳抑制效率的shRNA干扰载体。结果 Western blot法证实PLKO.1-STAT3-shRNA-1、2、3、4均能抑制细胞内STAT3基因的表达,以PLKO.1-STAT3-S1最为明显。用PLKO.1-STAT3-S1转染血管平滑肌细胞后可以促进细胞凋亡。结论成功构建了靶向STAT3基因的shRNA慢病毒干扰载体。
