AP-4(activator protein 4)是一种转录因子,在生物的生长发育中有重要作用。根据家蚕表达序列标签(EST)并利用cDNA末端快速扩增(RACE)方法克隆了家蚕AP-4(Bombyx mori activator protein4)基因,结合生物信息学方法对所获的序列进行开放...AP-4(activator protein 4)是一种转录因子,在生物的生长发育中有重要作用。根据家蚕表达序列标签(EST)并利用cDNA末端快速扩增(RACE)方法克隆了家蚕AP-4(Bombyx mori activator protein4)基因,结合生物信息学方法对所获的序列进行开放阅读框、序列同源性分析,预测了AP-4蛋白的理化性质。获得的家蚕AP-4基因cDNA的全长为1621bp,其开放阅读框为996bp,编码331个氨基酸,基因由3个外显子和2个内含子组成。同源性比对表明该基因推导的氨基酸序列与棉铃虫AP-4和谷蛀虫AP-4推测的蛋白同源性分别为76%和54%,第45-99位氨基酸序列是一个典型的保守结构域。实时定量RT-PCR显示该基因在所检测家蚕的各发育时期中,除幼虫1龄和蛹期第4天无表达外,其它时期均有表达,其中幼虫3龄和4龄期的表达量较高。展开更多
本研究利用生物信息学分析AP-4与胃癌患者临床病理信息的相关性,根据GenBank中人AP-4基因cDNA序列设计并合成特异性引物,以胃癌细胞总RNA逆转录的cDNA为模板,利用高保真酶扩增AP-4基因CDS (Coding DNA sequence)序列并构建入pcDNA3.1+载...本研究利用生物信息学分析AP-4与胃癌患者临床病理信息的相关性,根据GenBank中人AP-4基因cDNA序列设计并合成特异性引物,以胃癌细胞总RNA逆转录的cDNA为模板,利用高保真酶扩增AP-4基因CDS (Coding DNA sequence)序列并构建入pcDNA3.1+载体,并通过限制性内切酶酶切分析和测序法进行进一步验证;脂质体法将AP-4重组表达载体及对照pcDNA3.1+载体转染胃癌细胞,qRT-PCR(Quantitative real time polymerase chain reaction)和Western blotting检测分别检测AP-4在m RNA和蛋白水平的表达。生物信息学分析发现,AP-4的表达与胃癌分期及预后显著相关;酶切及测序分析表明,转录因子AP-4真核表达载体构建成功,并能够在胃癌细胞中实现转录和蛋白水平的高效表达。此研究为深入研究转录因子AP-4在胃癌等肿瘤发生发展中的作用及分子机制奠定了基础。展开更多
Proteolytic processing of the transmembrane amyloid precursor protein (APP) to aggregation-prone amyloid-β (Aβ) peptide underlies the development of Alzheimer’s disease.
Objective: To study the correlation of AP-4 and EZH2 gene expression with apoptosis and epithelial-mesenchymal transition in endometrial cancer lesions. Methods: Patients with endometrial cancer who received surgical ...Objective: To study the correlation of AP-4 and EZH2 gene expression with apoptosis and epithelial-mesenchymal transition in endometrial cancer lesions. Methods: Patients with endometrial cancer who received surgical resection in Xiaogan First People's Hospital between February 2015 and January 2017 were selected, right amount of endometrial cancer lesion and adjacent lesion was collected to extract RNA, and then the expression of AP-4 and EZH2 gene as well as apoptosis genes and epithelial-mesenchymal transition genes were determined. Results: AP-4 and EZH2 gene mRNA expression in endothelial cancer lesion were greatly higher than those in adjacent lesion;SRPX2, RLIP76, ZEB1, SALL4, TET1, RANKL and N-cadherin mRNA expression in endometrial cancer lesion were greatly higher than those in adjacent lesions whereas Fat-1, ITLN-1, Caspase-3 and -catenin mRNA expression were greatly lower than those in adjacent lesions;SRPX2 and RLIP76 mRNA expression in endometrial cancer lesion with high AP-4 expression were greatly higher than those in endometrial cancer lesion with low AP-4 expression whereas Fat-1, ITLN-1 and Caspase-3 mRNA expression were greatly lower than those in endometrial cancer lesion with low AP-4 expression;ZEB1, SALL4, TET1, RANKL and N-cadherin mRNA expression in endometrial cancer lesion with high EZH2 expression were greatly higher than those in endometrial cancer lesion with low EZH2 expression whereas α-catenin mRNA expression was greatly lower than that in endometrial cancer lesion with low EZH2 expression. Conclusion: The AP-4 and EZH2 gene highly expressed in endometrial cancer lesion can inhibit the apoptosis of cancer cells and promote the epithelial-mesenchymal transition of cancer cells.展开更多
Previous studies with deletion and sequence analysis of JDV LTR showed that there is a putative AP-4 responsive element in LTR. By antisense transient assay and gel shifting assay, we for the first time demonstrated t...Previous studies with deletion and sequence analysis of JDV LTR showed that there is a putative AP-4 responsive element in LTR. By antisense transient assay and gel shifting assay, we for the first time demonstrated that AP-4 modulated JDV gene expression by binding DNA directly to bovine cells. The results, derived from site-directed mutagenesis experiments, suggest that the six base pairs of AP-4 binding site (CAGCTG) have different effects on JDV gene expression. When the first two base pairs changed to GC, JDV gene expression is severely decreased.展开更多
Previous studies with deletion and sequence analysis of JDV LTR showed that there is a putative AP-4 responsive element in LTR. By antisense transient assay and gel shifting assay, we for the first time demonstrated t...Previous studies with deletion and sequence analysis of JDV LTR showed that there is a putative AP-4 responsive element in LTR. By antisense transient assay and gel shifting assay, we for the first time demonstrated that AP-4 modulated JDV gene expression by binding DNA di-rectly to bovine cells. The results, derived from site-directed mutagenesis experiments, suggest that the six base pairs of AP-4 binding site (CAGCTG) have different effects on JDV gene expression. When the first two base pairs changed to GC, JDV gene expression is severely decreased.展开更多
三叉神经痛(trigeminal neuralgia)是临床上的顽疾,长期以来为医学界所重视,但其发病机制尚不清晰,临床上也无有效的治疗方法。本研究首先通过免疫荧光化学法和动物行为学测定,对经眶下孔达三叉神经节目标注射给药法的效果进行了评价,...三叉神经痛(trigeminal neuralgia)是临床上的顽疾,长期以来为医学界所重视,但其发病机制尚不清晰,临床上也无有效的治疗方法。本研究首先通过免疫荧光化学法和动物行为学测定,对经眶下孔达三叉神经节目标注射给药法的效果进行了评价,然后行眶下神经慢性缩窄环术(chronic constriction injury of the infraorbital nerve,ION-CCI)建立三叉神经病理性痛大鼠模型,用经眶下孔达三叉神经节目标注射法分别注射BKCa通道激动剂NS1619和Kv通道拮抗剂4-AP,观察药物对大鼠面部机械痛阈的影响。结果显示,通过经眶下孔达三叉神经节目标注射法,药物可以准确到达三叉神经节,并产生了比一般眶下孔注射给药法更为持久的注射效果。ION-CCI术后第6天大鼠术侧面部触须垫部产生了显著的异常性疼痛(allodynia),并可以维持到至少术后第42天。在ION-CCI术后第15天大鼠,运用经眶下孔达三叉神经节目标注射法注射BKCa通道激动剂NS1619可以剂量依赖性地显著提高ION-CCI组大鼠的面部机械痛阈,逆转面部痛觉过敏;而在ION-CCI术后第35天痛阈部分恢复的大鼠,同样方法注射Kv通道拮抗剂4-AP又可以显著降低面部痛阈。以上结果表明,BKCa通道激动剂NS1619和Kv通道拮抗剂4-AP可以显著影响ION-CCI大鼠的面部机械痛阈,提示激活BKCa通道可以抑制由ION-CCI引起的三叉神经病理性痛,而激活Kv通道可能对ION-CCI引起的三叉神经病理痛有紧张性的抑制作用。展开更多
文摘AP-4(activator protein 4)是一种转录因子,在生物的生长发育中有重要作用。根据家蚕表达序列标签(EST)并利用cDNA末端快速扩增(RACE)方法克隆了家蚕AP-4(Bombyx mori activator protein4)基因,结合生物信息学方法对所获的序列进行开放阅读框、序列同源性分析,预测了AP-4蛋白的理化性质。获得的家蚕AP-4基因cDNA的全长为1621bp,其开放阅读框为996bp,编码331个氨基酸,基因由3个外显子和2个内含子组成。同源性比对表明该基因推导的氨基酸序列与棉铃虫AP-4和谷蛀虫AP-4推测的蛋白同源性分别为76%和54%,第45-99位氨基酸序列是一个典型的保守结构域。实时定量RT-PCR显示该基因在所检测家蚕的各发育时期中,除幼虫1龄和蛹期第4天无表达外,其它时期均有表达,其中幼虫3龄和4龄期的表达量较高。
文摘本研究利用生物信息学分析AP-4与胃癌患者临床病理信息的相关性,根据GenBank中人AP-4基因cDNA序列设计并合成特异性引物,以胃癌细胞总RNA逆转录的cDNA为模板,利用高保真酶扩增AP-4基因CDS (Coding DNA sequence)序列并构建入pcDNA3.1+载体,并通过限制性内切酶酶切分析和测序法进行进一步验证;脂质体法将AP-4重组表达载体及对照pcDNA3.1+载体转染胃癌细胞,qRT-PCR(Quantitative real time polymerase chain reaction)和Western blotting检测分别检测AP-4在m RNA和蛋白水平的表达。生物信息学分析发现,AP-4的表达与胃癌分期及预后显著相关;酶切及测序分析表明,转录因子AP-4真核表达载体构建成功,并能够在胃癌细胞中实现转录和蛋白水平的高效表达。此研究为深入研究转录因子AP-4在胃癌等肿瘤发生发展中的作用及分子机制奠定了基础。
文摘Proteolytic processing of the transmembrane amyloid precursor protein (APP) to aggregation-prone amyloid-β (Aβ) peptide underlies the development of Alzheimer’s disease.
文摘Objective: To study the correlation of AP-4 and EZH2 gene expression with apoptosis and epithelial-mesenchymal transition in endometrial cancer lesions. Methods: Patients with endometrial cancer who received surgical resection in Xiaogan First People's Hospital between February 2015 and January 2017 were selected, right amount of endometrial cancer lesion and adjacent lesion was collected to extract RNA, and then the expression of AP-4 and EZH2 gene as well as apoptosis genes and epithelial-mesenchymal transition genes were determined. Results: AP-4 and EZH2 gene mRNA expression in endothelial cancer lesion were greatly higher than those in adjacent lesion;SRPX2, RLIP76, ZEB1, SALL4, TET1, RANKL and N-cadherin mRNA expression in endometrial cancer lesion were greatly higher than those in adjacent lesions whereas Fat-1, ITLN-1, Caspase-3 and -catenin mRNA expression were greatly lower than those in adjacent lesions;SRPX2 and RLIP76 mRNA expression in endometrial cancer lesion with high AP-4 expression were greatly higher than those in endometrial cancer lesion with low AP-4 expression whereas Fat-1, ITLN-1 and Caspase-3 mRNA expression were greatly lower than those in endometrial cancer lesion with low AP-4 expression;ZEB1, SALL4, TET1, RANKL and N-cadherin mRNA expression in endometrial cancer lesion with high EZH2 expression were greatly higher than those in endometrial cancer lesion with low EZH2 expression whereas α-catenin mRNA expression was greatly lower than that in endometrial cancer lesion with low EZH2 expression. Conclusion: The AP-4 and EZH2 gene highly expressed in endometrial cancer lesion can inhibit the apoptosis of cancer cells and promote the epithelial-mesenchymal transition of cancer cells.
基金国家自然科学基金，the High Educational Outstanding Teachers Help Plan
文摘Previous studies with deletion and sequence analysis of JDV LTR showed that there is a putative AP-4 responsive element in LTR. By antisense transient assay and gel shifting assay, we for the first time demonstrated that AP-4 modulated JDV gene expression by binding DNA directly to bovine cells. The results, derived from site-directed mutagenesis experiments, suggest that the six base pairs of AP-4 binding site (CAGCTG) have different effects on JDV gene expression. When the first two base pairs changed to GC, JDV gene expression is severely decreased.
文摘Previous studies with deletion and sequence analysis of JDV LTR showed that there is a putative AP-4 responsive element in LTR. By antisense transient assay and gel shifting assay, we for the first time demonstrated that AP-4 modulated JDV gene expression by binding DNA di-rectly to bovine cells. The results, derived from site-directed mutagenesis experiments, suggest that the six base pairs of AP-4 binding site (CAGCTG) have different effects on JDV gene expression. When the first two base pairs changed to GC, JDV gene expression is severely decreased.
基金supported by the Basic Research Program of Science and Technology Commission of Shanghai MunicipalityChina(No.08JC1405400)
文摘三叉神经痛(trigeminal neuralgia)是临床上的顽疾,长期以来为医学界所重视,但其发病机制尚不清晰,临床上也无有效的治疗方法。本研究首先通过免疫荧光化学法和动物行为学测定,对经眶下孔达三叉神经节目标注射给药法的效果进行了评价,然后行眶下神经慢性缩窄环术(chronic constriction injury of the infraorbital nerve,ION-CCI)建立三叉神经病理性痛大鼠模型,用经眶下孔达三叉神经节目标注射法分别注射BKCa通道激动剂NS1619和Kv通道拮抗剂4-AP,观察药物对大鼠面部机械痛阈的影响。结果显示,通过经眶下孔达三叉神经节目标注射法,药物可以准确到达三叉神经节,并产生了比一般眶下孔注射给药法更为持久的注射效果。ION-CCI术后第6天大鼠术侧面部触须垫部产生了显著的异常性疼痛(allodynia),并可以维持到至少术后第42天。在ION-CCI术后第15天大鼠,运用经眶下孔达三叉神经节目标注射法注射BKCa通道激动剂NS1619可以剂量依赖性地显著提高ION-CCI组大鼠的面部机械痛阈,逆转面部痛觉过敏;而在ION-CCI术后第35天痛阈部分恢复的大鼠,同样方法注射Kv通道拮抗剂4-AP又可以显著降低面部痛阈。以上结果表明,BKCa通道激动剂NS1619和Kv通道拮抗剂4-AP可以显著影响ION-CCI大鼠的面部机械痛阈,提示激活BKCa通道可以抑制由ION-CCI引起的三叉神经病理性痛,而激活Kv通道可能对ION-CCI引起的三叉神经病理痛有紧张性的抑制作用。
