Objective:To investigate the changes of Numb and TFR1 expression in ovarian cancer tissue and their clinical pathological value.Methods:90 patients with ovarian cancer underwent radical operation in Pidu District Peop...Objective:To investigate the changes of Numb and TFR1 expression in ovarian cancer tissue and their clinical pathological value.Methods:90 patients with ovarian cancer underwent radical operation in Pidu District People's Hospital,the First Affiliated Hospital of Chengdu Medical College and Mianyang Central Hospital between August 2015 and April 2018,115 patients with ovarian chocolate cyst underwent surgery during the same period,and intraoperative lesion tissue samples of the two groups were collected and enrolled in ovarian cancer group and ovarian chocolate cyst group respectively.The expressions levels of Numb,TFR1 gene as well as ovarian cancer-related proliferation,invasion and autophagy genes in ovarian lesions were detected.Pearson test was used to evaluate the correlation of Numb and TFR1 gene expression in ovarian cancer tissues with proliferation,invasion and autophagy activity of cancer cells.Results:Numb mRNA expression in lesions of ovarian cancer group was lower than that of ovarian chocolate cyst group whereas TFR1 mRNA expression was higher than that of ovarian chocolate cyst group;proliferation genes C3G,Clusterin,FUNDC1,PIWIL4 and TNFAIP8 mRNA expression were higher than those of ovarian chocolate cyst group whereas Let-7i mRNA expression was lower than that of ovarian chocolate cyst group;invasion genes BRMS1,XAV939 and OVCA1 mRMA expression were lower than those of ovarian chocolate cyst group whereas CUL4A and MTA1 mRNA expression were higher than those of ovarian chocolate cyst group;autophagy genes FTY720 and Beclin1 mRNA expression were lower than those of ovarian chocolate cyst group whereas NAC-1 mRNA expression was higher than that of ovarian chocolate cyst group.Correlation analysis showed that the Numb and TFR1 gene expression in ovarian cancer tissues were directly correlated with the proliferation,invasion and autophagy activity of cancer cells.Conclusion:The abnormal decrease of Numb gene expression and the abnormal increase of TFR1 gene expression in ovarian cancer tissues are directly correlated with the malignant biological behaviors of cancer cells.展开更多
目的开发靶向转铁蛋白受体1(transferrin receptor 1,TfR1)且具有跨血脑屏障转运功能的鲨鱼纳米抗体。方法提取未经靶抗原免疫的条纹斑竹鲨外周血单个核细胞总RNA,通过PCR技术扩增获得鲨鱼抗体的重链可变区(variable domain of immunogl...目的开发靶向转铁蛋白受体1(transferrin receptor 1,TfR1)且具有跨血脑屏障转运功能的鲨鱼纳米抗体。方法提取未经靶抗原免疫的条纹斑竹鲨外周血单个核细胞总RNA,通过PCR技术扩增获得鲨鱼抗体的重链可变区(variable domain of immunoglobulin new antigen receptor,VNAR)基因,将VNAR构建至噬菌体展示载体中,完成鲨鱼纳米抗体天然文库的构建。扩增天然噬菌体展示文库,采用抗原固相淘选策略,通过三轮“结合-洗脱-扩增”的淘选流程完成噬菌体展示文库的富集,通过噬菌体ELISA鉴定阳性克隆,并进行抗体序列比对,以确定靶向TfR1的鲨鱼纳米抗体序列。构建鲨鱼纳米抗体重组表达载体,利用大肠杆菌原核表达系统进行鲨鱼纳米抗体的诱导表达,ELISA检测鲨鱼纳米抗体与TfR1重组蛋白的结合能力,竞争性ELISA检测鲨鱼纳米抗体与转铁蛋白(transferrin,Tf)竞争受体结合表位的情况,免疫荧光技术检测鲨鱼纳米抗体与脑微血管内皮细胞bEnd.3的结合能力,血脑屏障体外实验模型检测鲨鱼纳米抗体的跨血脑屏障转运能力。结果成功构建了库容量为2.35×10^(9) CFU的鲨鱼纳米抗体天然文库,该文库具备良好的多样性。经淘选获得2株靶向TfR1的鲨鱼纳米抗体,均具有亲和活性,其中D1具有跨血脑屏障转运功能。结论本研究成功筛选出靶向TfR1且具有跨血脑屏障转运功能的鲨鱼纳米抗体,为中枢神经系统疾病治疗提供了新的跨血脑屏障药物递送载体。展开更多
通过将体内稳定同位素标记细胞培养技术(AACT/SILAC)与LC-MS/MS联用,对哺乳动物细胞中TNF-α/NF-κB信号通路相关蛋白在TNF-α及TSA存在情况下乙酰化变化情况进行了研究.在TSA协同处理的、TNF-α刺激的细胞中鉴定到3个乙酰化蛋白,其中...通过将体内稳定同位素标记细胞培养技术(AACT/SILAC)与LC-MS/MS联用,对哺乳动物细胞中TNF-α/NF-κB信号通路相关蛋白在TNF-α及TSA存在情况下乙酰化变化情况进行了研究.在TSA协同处理的、TNF-α刺激的细胞中鉴定到3个乙酰化蛋白,其中核糖体蛋白L4(60SRibosomal Protein L4,RPL4)已被报道存在乙酰化的后修饰,转铁蛋白受体1(Transferrin Receptor Protein 1,TfR1)和凋亡诱导因子3(ApoptosisInducing Factor,Mitochondrion-associated,3,AIFM3)尚未见报道.展开更多
基金Research Projects of Health and Family Planning Commission of Sichuan Province.Project No:110530.
文摘Objective:To investigate the changes of Numb and TFR1 expression in ovarian cancer tissue and their clinical pathological value.Methods:90 patients with ovarian cancer underwent radical operation in Pidu District People's Hospital,the First Affiliated Hospital of Chengdu Medical College and Mianyang Central Hospital between August 2015 and April 2018,115 patients with ovarian chocolate cyst underwent surgery during the same period,and intraoperative lesion tissue samples of the two groups were collected and enrolled in ovarian cancer group and ovarian chocolate cyst group respectively.The expressions levels of Numb,TFR1 gene as well as ovarian cancer-related proliferation,invasion and autophagy genes in ovarian lesions were detected.Pearson test was used to evaluate the correlation of Numb and TFR1 gene expression in ovarian cancer tissues with proliferation,invasion and autophagy activity of cancer cells.Results:Numb mRNA expression in lesions of ovarian cancer group was lower than that of ovarian chocolate cyst group whereas TFR1 mRNA expression was higher than that of ovarian chocolate cyst group;proliferation genes C3G,Clusterin,FUNDC1,PIWIL4 and TNFAIP8 mRNA expression were higher than those of ovarian chocolate cyst group whereas Let-7i mRNA expression was lower than that of ovarian chocolate cyst group;invasion genes BRMS1,XAV939 and OVCA1 mRMA expression were lower than those of ovarian chocolate cyst group whereas CUL4A and MTA1 mRNA expression were higher than those of ovarian chocolate cyst group;autophagy genes FTY720 and Beclin1 mRNA expression were lower than those of ovarian chocolate cyst group whereas NAC-1 mRNA expression was higher than that of ovarian chocolate cyst group.Correlation analysis showed that the Numb and TFR1 gene expression in ovarian cancer tissues were directly correlated with the proliferation,invasion and autophagy activity of cancer cells.Conclusion:The abnormal decrease of Numb gene expression and the abnormal increase of TFR1 gene expression in ovarian cancer tissues are directly correlated with the malignant biological behaviors of cancer cells.
文摘目的开发靶向转铁蛋白受体1(transferrin receptor 1,TfR1)且具有跨血脑屏障转运功能的鲨鱼纳米抗体。方法提取未经靶抗原免疫的条纹斑竹鲨外周血单个核细胞总RNA,通过PCR技术扩增获得鲨鱼抗体的重链可变区(variable domain of immunoglobulin new antigen receptor,VNAR)基因,将VNAR构建至噬菌体展示载体中,完成鲨鱼纳米抗体天然文库的构建。扩增天然噬菌体展示文库,采用抗原固相淘选策略,通过三轮“结合-洗脱-扩增”的淘选流程完成噬菌体展示文库的富集,通过噬菌体ELISA鉴定阳性克隆,并进行抗体序列比对,以确定靶向TfR1的鲨鱼纳米抗体序列。构建鲨鱼纳米抗体重组表达载体,利用大肠杆菌原核表达系统进行鲨鱼纳米抗体的诱导表达,ELISA检测鲨鱼纳米抗体与TfR1重组蛋白的结合能力,竞争性ELISA检测鲨鱼纳米抗体与转铁蛋白(transferrin,Tf)竞争受体结合表位的情况,免疫荧光技术检测鲨鱼纳米抗体与脑微血管内皮细胞bEnd.3的结合能力,血脑屏障体外实验模型检测鲨鱼纳米抗体的跨血脑屏障转运能力。结果成功构建了库容量为2.35×10^(9) CFU的鲨鱼纳米抗体天然文库,该文库具备良好的多样性。经淘选获得2株靶向TfR1的鲨鱼纳米抗体,均具有亲和活性,其中D1具有跨血脑屏障转运功能。结论本研究成功筛选出靶向TfR1且具有跨血脑屏障转运功能的鲨鱼纳米抗体,为中枢神经系统疾病治疗提供了新的跨血脑屏障药物递送载体。
文摘通过将体内稳定同位素标记细胞培养技术(AACT/SILAC)与LC-MS/MS联用,对哺乳动物细胞中TNF-α/NF-κB信号通路相关蛋白在TNF-α及TSA存在情况下乙酰化变化情况进行了研究.在TSA协同处理的、TNF-α刺激的细胞中鉴定到3个乙酰化蛋白,其中核糖体蛋白L4(60SRibosomal Protein L4,RPL4)已被报道存在乙酰化的后修饰,转铁蛋白受体1(Transferrin Receptor Protein 1,TfR1)和凋亡诱导因子3(ApoptosisInducing Factor,Mitochondrion-associated,3,AIFM3)尚未见报道.
