为探究一种包含泛素调节性X结构域的蛋白(ubiquitin regulatory X domain-containing protein,UBXN1)在大黄鱼抗盾纤毛虫感染中的作用,以及可能涉及的免疫信号通路。本实验克隆鉴定了大黄鱼UBXN1基因,并利用在线软件对其序列特征进行生...为探究一种包含泛素调节性X结构域的蛋白(ubiquitin regulatory X domain-containing protein,UBXN1)在大黄鱼抗盾纤毛虫感染中的作用,以及可能涉及的免疫信号通路。本实验克隆鉴定了大黄鱼UBXN1基因,并利用在线软件对其序列特征进行生物信息学分析;采用实时荧光定量PCR(qRT-PCR)检测UBXN1在健康大黄鱼各组织中的表达,及盾纤毛虫感染后的诱导表达变化;并进行了UBXN1的亚细胞定位;转录组测序分析了UBXN1过表达前后的差异表达基因。结果显示,UBXN1基因cDNA全长为915 bp,编码304个氨基酸。蛋白多重序列比对和结构预测表明UBXN1是一个进化保守的蛋白,包含UBA和UBX结构域。qRT-PCR分析表明UBXN1在所检测的11种组织中均有表达,脑中表达量最高,其次是肝脏、心脏和肾脏,在肌肉中表达量最低;盾纤毛虫感染大黄鱼后,UBXN1在脾脏、脑、肝脏和肾脏中表达量早期显著升高,后期逐步恢复至正常水平。亚细胞定位分析表明,UBXN1在大黄鱼肾脏细胞质和细胞核中均有表达。在293T细胞过表达UBXN1,转录组差异表达分析筛选到12个上调基因,4个下调基因,其中RPL41/RPL39/XIST/RNA45SN4表达量显著增加,而ATP8/ND4L表达量显著减少。研究表明UBXN1在大黄鱼抗寄生虫免疫应答中发挥重要作用。本实验为进一步研究UBXN1的免疫信号通路奠定基础。展开更多
Both hepatitis B virus X protein(HBx)and microRNA-221(miR-221)have been implicated in the development of hepatitis B virus(HBV)-related hepatocellular carcinoma(HCC).The present study demonstrates that HBx promotes HC...Both hepatitis B virus X protein(HBx)and microRNA-221(miR-221)have been implicated in the development of hepatitis B virus(HBV)-related hepatocellular carcinoma(HCC).The present study demonstrates that HBx promotes HCC cell proliferation via the C-X-C motif chemokine ligand 12-C-X-C chemokine receptor type 4(CXCL12-CXCR4)axis.We predict that HBx/miR-221-mediated CXCL12/CXCR4 signaling induces NKT cells to promote HBV-related HCC.Methods:After miR-221 mimic,miR-221 mimic negative control,miR-221 inhibitor,miR-221 inhibitor negative control were transfected into cells,the expression of CXCL12 and miR-221 was detected by qPCR and western blot.Then we constructed a stable HBV-HCC cell line.HBV-HCC cells were injected into the nude mice,thus a HBV-HCC mouse model was constructed.Q-PCR and western blot were used to detect the expression of HBx,miR-221,CXCL12 and CXCR4 in tumor tissues.The expression of CXCL12 was detected by immunohistochemistry,and the expression of CXCR4,CD3 and CD56 was detected by immunofluorescence.The levels of CXCL12,IL-2 and TNF-αin serum of mice were detected by ELISA.Sixty-one patients with HBV-related HCC,61 patients with HBV-related cirrhosis,61 patients with chronic hepatitis B(CHB)and 30 healthy people were enrolled.CXCL12,cytokine levels,and clinicopathological parameters were tested.Results:Hepatitis B virus X protein upregulates the expression of miR-221 and CXCL12 in lentivirus(LV5)-HBx-transfected HepG2 cells.HBx protein promotes HepG2 cell proliferation in vitro.HBx protein promoted tumor growth via the miR-221/CXCL12/CXCR4 pathway in a mouse tumor model.HBx protein upregulated natural killer T cell expression via the CXCR4/CXCL12 pathway to promote tumor growth.The data demonstrated a positive correlation between CXCL12 concentration with Cre levels and Child-Pugh scores.CXCL12 had an inferior diagnostic efficiency compared to IL-2 and IL-6 for HBV-related HCC.Conclusions:We present evidence that HBx/miR-221-mediated CXCL12/CXCR4 signaling induces NKT cells to promote HBV-related HCC.展开更多
脆性X综合征(fragile X syndrome,FXS)是FMR1基因CGG异常重复扩增导致的疾病。本文报告1对经基因检测诊断为FXS的兄弟,2例患者分别为15岁和14岁,均存在语言障碍、智力障碍、注意力缺陷障碍、孤独症谱系障碍和FXS特征性面容等临床表现,...脆性X综合征(fragile X syndrome,FXS)是FMR1基因CGG异常重复扩增导致的疾病。本文报告1对经基因检测诊断为FXS的兄弟,2例患者分别为15岁和14岁,均存在语言障碍、智力障碍、注意力缺陷障碍、孤独症谱系障碍和FXS特征性面容等临床表现,其中先证者伴有罕见的晚发性癫痫发作,经左乙拉西坦治疗效果良好,而其弟弟经反复随访未见脑电图异常。该对病例提示FXS临床表型具有多样性和异质性。展开更多
目的·分析癌-睾丸抗原(cancer-testis antigen,CTA)家族成员SPANXB(sperm protein associated with the nucleus on the X chromosome B)在肝癌中的表达及其与肝癌患者预后之间的相关性,并探究SPANXB对肝癌细胞增殖的影响及其潜在...目的·分析癌-睾丸抗原(cancer-testis antigen,CTA)家族成员SPANXB(sperm protein associated with the nucleus on the X chromosome B)在肝癌中的表达及其与肝癌患者预后之间的相关性,并探究SPANXB对肝癌细胞增殖的影响及其潜在机制。方法·利用癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中的肝癌样本数据,分析SPANXB在肝癌组织中的表达及其与患者生存期的相关性。构建稳定敲低SPANXB与稳定过表达SPANXB的肝癌细胞系,利用活细胞成像实验、EdU细胞增殖实验和平板克隆形成实验评估SPANXB对肝癌细胞增殖的影响。通过RNA测序(RNA-sequence,RNA-seq)探究SPANXB调控肝癌细胞增殖的相关通路,并利用细胞周期实验验证SPANXB对肝癌细胞周期的影响。采用免疫沉淀-质谱联用技术(immunoprecipitation-mass spectrometry,IP-MS)探索与SPANXB相互作用的蛋白,并使用免疫共沉淀(co-immunoprecipitation,Co-IP)进行验证。结果·SPANXB mRNA在肝癌组织中的表达高于正常组织(P=0.003),且与肝癌患者的生存期呈负相关。稳定敲低SPANXB可降低肝癌细胞的增殖能力、克隆形成能力,而稳定过表达SPANXB则可促进这些过程。RNA-seq的结果显示,SPANXB的敲低可下调DNA复制与G1/S细胞周期转换相关通路,细胞周期实验的结果显示SPANXB的敲低可导致肝癌细胞周期发生改变。IP-MS和Co-IP结果显示,SPAXNB与有丝分裂停滞缺陷2样蛋白1(mitotic arrest deficient 2-like protein 1,MAD2L1)、WD重复域蛋白5(WD repeat domain 5,WDR5)等细胞周期相关蛋白存在相互作用。结论·SPANXB的高表达与肝癌的预后呈负相关,其可能通过与MAD2L1、WDR5相互作用调控细胞周期并增强肝癌细胞的增殖活性。展开更多
基金supported by the National Natural Science Foundation of Chongqing,China(No.cstc2019jcyj-msxm0314 of Yishu Tang)the National Key R&D Program of China(No.2017YFC0909902 of Yun Xia)the Natural Science Foundation of China(No.81501818 of Yishu Tang)。
文摘Both hepatitis B virus X protein(HBx)and microRNA-221(miR-221)have been implicated in the development of hepatitis B virus(HBV)-related hepatocellular carcinoma(HCC).The present study demonstrates that HBx promotes HCC cell proliferation via the C-X-C motif chemokine ligand 12-C-X-C chemokine receptor type 4(CXCL12-CXCR4)axis.We predict that HBx/miR-221-mediated CXCL12/CXCR4 signaling induces NKT cells to promote HBV-related HCC.Methods:After miR-221 mimic,miR-221 mimic negative control,miR-221 inhibitor,miR-221 inhibitor negative control were transfected into cells,the expression of CXCL12 and miR-221 was detected by qPCR and western blot.Then we constructed a stable HBV-HCC cell line.HBV-HCC cells were injected into the nude mice,thus a HBV-HCC mouse model was constructed.Q-PCR and western blot were used to detect the expression of HBx,miR-221,CXCL12 and CXCR4 in tumor tissues.The expression of CXCL12 was detected by immunohistochemistry,and the expression of CXCR4,CD3 and CD56 was detected by immunofluorescence.The levels of CXCL12,IL-2 and TNF-αin serum of mice were detected by ELISA.Sixty-one patients with HBV-related HCC,61 patients with HBV-related cirrhosis,61 patients with chronic hepatitis B(CHB)and 30 healthy people were enrolled.CXCL12,cytokine levels,and clinicopathological parameters were tested.Results:Hepatitis B virus X protein upregulates the expression of miR-221 and CXCL12 in lentivirus(LV5)-HBx-transfected HepG2 cells.HBx protein promotes HepG2 cell proliferation in vitro.HBx protein promoted tumor growth via the miR-221/CXCL12/CXCR4 pathway in a mouse tumor model.HBx protein upregulated natural killer T cell expression via the CXCR4/CXCL12 pathway to promote tumor growth.The data demonstrated a positive correlation between CXCL12 concentration with Cre levels and Child-Pugh scores.CXCL12 had an inferior diagnostic efficiency compared to IL-2 and IL-6 for HBV-related HCC.Conclusions:We present evidence that HBx/miR-221-mediated CXCL12/CXCR4 signaling induces NKT cells to promote HBV-related HCC.
文摘脆性X综合征(fragile X syndrome,FXS)是FMR1基因CGG异常重复扩增导致的疾病。本文报告1对经基因检测诊断为FXS的兄弟,2例患者分别为15岁和14岁,均存在语言障碍、智力障碍、注意力缺陷障碍、孤独症谱系障碍和FXS特征性面容等临床表现,其中先证者伴有罕见的晚发性癫痫发作,经左乙拉西坦治疗效果良好,而其弟弟经反复随访未见脑电图异常。该对病例提示FXS临床表型具有多样性和异质性。
文摘目的·分析癌-睾丸抗原(cancer-testis antigen,CTA)家族成员SPANXB(sperm protein associated with the nucleus on the X chromosome B)在肝癌中的表达及其与肝癌患者预后之间的相关性,并探究SPANXB对肝癌细胞增殖的影响及其潜在机制。方法·利用癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中的肝癌样本数据,分析SPANXB在肝癌组织中的表达及其与患者生存期的相关性。构建稳定敲低SPANXB与稳定过表达SPANXB的肝癌细胞系,利用活细胞成像实验、EdU细胞增殖实验和平板克隆形成实验评估SPANXB对肝癌细胞增殖的影响。通过RNA测序(RNA-sequence,RNA-seq)探究SPANXB调控肝癌细胞增殖的相关通路,并利用细胞周期实验验证SPANXB对肝癌细胞周期的影响。采用免疫沉淀-质谱联用技术(immunoprecipitation-mass spectrometry,IP-MS)探索与SPANXB相互作用的蛋白,并使用免疫共沉淀(co-immunoprecipitation,Co-IP)进行验证。结果·SPANXB mRNA在肝癌组织中的表达高于正常组织(P=0.003),且与肝癌患者的生存期呈负相关。稳定敲低SPANXB可降低肝癌细胞的增殖能力、克隆形成能力,而稳定过表达SPANXB则可促进这些过程。RNA-seq的结果显示,SPANXB的敲低可下调DNA复制与G1/S细胞周期转换相关通路,细胞周期实验的结果显示SPANXB的敲低可导致肝癌细胞周期发生改变。IP-MS和Co-IP结果显示,SPAXNB与有丝分裂停滞缺陷2样蛋白1(mitotic arrest deficient 2-like protein 1,MAD2L1)、WD重复域蛋白5(WD repeat domain 5,WDR5)等细胞周期相关蛋白存在相互作用。结论·SPANXB的高表达与肝癌的预后呈负相关,其可能通过与MAD2L1、WDR5相互作用调控细胞周期并增强肝癌细胞的增殖活性。