目的探讨肿瘤坏死因子受体相关因子-2和Nck相互作用蛋白激酶(Traf-2 and Nck-interacting protein kinase,TNIK)在胃癌组织中的表达及其临床意义。方法回顾性收集2014年7月至2017年12月期间在郑州大学附属洛阳中心医院普通外科行手术切...目的探讨肿瘤坏死因子受体相关因子-2和Nck相互作用蛋白激酶(Traf-2 and Nck-interacting protein kinase,TNIK)在胃癌组织中的表达及其临床意义。方法回顾性收集2014年7月至2017年12月期间在郑州大学附属洛阳中心医院普通外科行手术切除并经病理科确诊的胃癌组织及其癌旁组织标本78份,以及同期胃溃疡组织标本42份。采用免疫组织化学染色法及Western blot法检测上述3种组织中TNIK蛋白的表达情况,并探索TNIK蛋白表达与胃癌临床病理学特征及预后的关系。结果免疫组织化学染色结果显示,胃癌组中,TNIK蛋白的阳性表达率为66.67%(52/78),癌旁组织组为11.54%(9/78),胃溃疡组为21.43%(9/42)。胃癌组的TNIK蛋白阳性表达率高于癌旁组织组和胃溃疡组(P<0.05),但癌旁组织组和胃溃疡组比较差异无统计学意义(P>0.05)。Western blot结果显示,在胃癌组中,TNIK蛋白的表达水平高于胃溃疡组及癌旁组织组,差异均有统计学意义(P<0.05);但癌旁组织组与胃溃疡组比较差异无统计学意义(P=0.772)。TNIK蛋白的表达与胃癌患者的性别、CEA值、肿瘤直径及病理学类型均无关(P>0.05),但与胃癌患者的年龄、TNM分期、分化程度、远处转移及淋巴结转移相关(P<0.05);TNIK蛋白阳性表达患者的预后差于阴性表达者(P<0.001)。结论TNIK蛋白在胃癌组织中高表达,且与胃癌预后不良因素及较差的预后相关,可能是潜在的胃癌诊治靶点。展开更多
Epilepsy,one of the most common neurological disorders,is characterized by spon-taneous recurrent seizures.Temporal lobe epilepsy(TLE)is one of the most common medically intractable seizure disorders.Traf2-and NcK-int...Epilepsy,one of the most common neurological disorders,is characterized by spon-taneous recurrent seizures.Temporal lobe epilepsy(TLE)is one of the most common medically intractable seizure disorders.Traf2-and NcK-interacting kinase(TNIK)has recently attracted attention as a critical modulation target of many neurological and psychiatric disorders,but its role in epilepsy remains unclear.In this study,we hypothesized the involvement of TNIK in epilepsy and investigated TNIK expression in patients with intractable TLE and in a pilocarpineinduced rat model of epilepsy by western blotting,immunofluorescence,and immunohistochemistry.A pentylenetetrazole(PTZ)-induced epilepsy rat model was used to determine the effect of the TNIK inhibitor NCB-0846 on behavioral manifestations of epilepsy.Coimmunoprecipitation(Co-IP)/mass spectrometry(MS)was used to identify the potential mechanism.Through Co-IP,we detected and confirmed the main potential TNIK interactors.Subcellular fractionation was used to establish the effect of NCB-0846 on the expression of the main interactors in postsynaptic density(PSD)fractions.We found that TNIK was primarily located in neurons and decreased significantly in epilepsy model rats and TLE patients compared with controls.NCB-0846 delayed kindling progression and decreased seizure severity.Co-IP/MS identified 63 candidate TNIK interactors in rat hippocampi,notably CaMKIl.Co-IP showed that TNIK might correlate with endogenous GRIA1,SYN2,PSD-95,CaMKIV,GABRG1,and GABRG2.In addition,the significant decrease in GRIA1 in hippocampal total lysate and PSDs after NCB-0846 treatment might help modify the progression of PTZ kindling.Our results suggest that TNIK contributes to epileptic pathology and is a potential antiepileptic drug target.展开更多
AIM： To find the relationship between hepatitis B virus （HBV） and hepatocytes during the initial state of infection by cDNA microarray. METHODS： Primary normal human hepatocytes （PNHHs） were isolated and infecte...AIM： To find the relationship between hepatitis B virus （HBV） and hepatocytes during the initial state of infection by cDNA microarray. METHODS： Primary normal human hepatocytes （PNHHs） were isolated and infected with HBV. From the PNHHs, RNA was isolated and inverted into complement DNA （cDNA） with Cy3- or Cy5- labeled dUTP for microarray analysis. The labeled cDNA was hybridized with microarray chip, including 4224 cDNAs. From the image of the microarray, expression profiles were produced and some of them were confirmed by RT-PCR, immunoblot analysis, and NF-κB luciferase reporter assay. RESULTS： From the cDNA microarray, we obtained 98 differentially regulated genes. Of the 98 genes, 53 were up regulated and 45 down regulated. Interestingly, in the up regulated genes, we found the TNF signaling pathway-related genes： LT-α, TRAF2, and NIK. By using RT-PCR, we confirmed the up-regulation of these genes in HepG2, HuhT, and Chang liver cells, which were transfected with pHBV1.2x, a plasmid encoding all HBV messages. Moreover, these three genes participated in HBV- mediated NF-κB activation. CONCLUSION： During the initial state of HBV infection, hepatocytes facilitate the activation of NF-κB through up regulation of LT-α, TRAF2, and NIK.展开更多
文摘目的探讨肿瘤坏死因子受体相关因子-2和Nck相互作用蛋白激酶(Traf-2 and Nck-interacting protein kinase,TNIK)在胃癌组织中的表达及其临床意义。方法回顾性收集2014年7月至2017年12月期间在郑州大学附属洛阳中心医院普通外科行手术切除并经病理科确诊的胃癌组织及其癌旁组织标本78份,以及同期胃溃疡组织标本42份。采用免疫组织化学染色法及Western blot法检测上述3种组织中TNIK蛋白的表达情况,并探索TNIK蛋白表达与胃癌临床病理学特征及预后的关系。结果免疫组织化学染色结果显示,胃癌组中,TNIK蛋白的阳性表达率为66.67%(52/78),癌旁组织组为11.54%(9/78),胃溃疡组为21.43%(9/42)。胃癌组的TNIK蛋白阳性表达率高于癌旁组织组和胃溃疡组(P<0.05),但癌旁组织组和胃溃疡组比较差异无统计学意义(P>0.05)。Western blot结果显示,在胃癌组中,TNIK蛋白的表达水平高于胃溃疡组及癌旁组织组,差异均有统计学意义(P<0.05);但癌旁组织组与胃溃疡组比较差异无统计学意义(P=0.772)。TNIK蛋白的表达与胃癌患者的性别、CEA值、肿瘤直径及病理学类型均无关(P>0.05),但与胃癌患者的年龄、TNM分期、分化程度、远处转移及淋巴结转移相关(P<0.05);TNIK蛋白阳性表达患者的预后差于阴性表达者(P<0.001)。结论TNIK蛋白在胃癌组织中高表达,且与胃癌预后不良因素及较差的预后相关,可能是潜在的胃癌诊治靶点。
基金supported by grants from the National Natural Science Foundation of China(No.81901324,82001378,82071395)China Postdoctoral Science Foundation(No.2021M693246)+4 种基金joint project of Chongqing Health Commission and Science and Technology Bureau(Chongqing,China)(No.2023QNXM009)Science and Technology Research Program of Chongqing Education Commission of China(No.KJQN202200435)Chongqing Talents:Exceptional Young Talents Project(Chongqing,China)(No.CQYC202005014)Natural Science Foundation of Chongqing,China(No.cstc2021ycjh-bgzxm0035,CSTB2022NSCQ-LZX0038)Key Research and Development Projects of Jining City,Shandong,China(No.2021YXNS057).
文摘Epilepsy,one of the most common neurological disorders,is characterized by spon-taneous recurrent seizures.Temporal lobe epilepsy(TLE)is one of the most common medically intractable seizure disorders.Traf2-and NcK-interacting kinase(TNIK)has recently attracted attention as a critical modulation target of many neurological and psychiatric disorders,but its role in epilepsy remains unclear.In this study,we hypothesized the involvement of TNIK in epilepsy and investigated TNIK expression in patients with intractable TLE and in a pilocarpineinduced rat model of epilepsy by western blotting,immunofluorescence,and immunohistochemistry.A pentylenetetrazole(PTZ)-induced epilepsy rat model was used to determine the effect of the TNIK inhibitor NCB-0846 on behavioral manifestations of epilepsy.Coimmunoprecipitation(Co-IP)/mass spectrometry(MS)was used to identify the potential mechanism.Through Co-IP,we detected and confirmed the main potential TNIK interactors.Subcellular fractionation was used to establish the effect of NCB-0846 on the expression of the main interactors in postsynaptic density(PSD)fractions.We found that TNIK was primarily located in neurons and decreased significantly in epilepsy model rats and TLE patients compared with controls.NCB-0846 delayed kindling progression and decreased seizure severity.Co-IP/MS identified 63 candidate TNIK interactors in rat hippocampi,notably CaMKIl.Co-IP showed that TNIK might correlate with endogenous GRIA1,SYN2,PSD-95,CaMKIV,GABRG1,and GABRG2.In addition,the significant decrease in GRIA1 in hippocampal total lysate and PSDs after NCB-0846 treatment might help modify the progression of PTZ kindling.Our results suggest that TNIK contributes to epileptic pathology and is a potential antiepileptic drug target.
基金Supported by a grant of the Korea Health 21 R&D Project, Ministry of Health and Welfare, Republic of Korea, No. A050145
文摘AIM： To find the relationship between hepatitis B virus （HBV） and hepatocytes during the initial state of infection by cDNA microarray. METHODS： Primary normal human hepatocytes （PNHHs） were isolated and infected with HBV. From the PNHHs, RNA was isolated and inverted into complement DNA （cDNA） with Cy3- or Cy5- labeled dUTP for microarray analysis. The labeled cDNA was hybridized with microarray chip, including 4224 cDNAs. From the image of the microarray, expression profiles were produced and some of them were confirmed by RT-PCR, immunoblot analysis, and NF-κB luciferase reporter assay. RESULTS： From the cDNA microarray, we obtained 98 differentially regulated genes. Of the 98 genes, 53 were up regulated and 45 down regulated. Interestingly, in the up regulated genes, we found the TNF signaling pathway-related genes： LT-α, TRAF2, and NIK. By using RT-PCR, we confirmed the up-regulation of these genes in HepG2, HuhT, and Chang liver cells, which were transfected with pHBV1.2x, a plasmid encoding all HBV messages. Moreover, these three genes participated in HBV- mediated NF-κB activation. CONCLUSION： During the initial state of HBV infection, hepatocytes facilitate the activation of NF-κB through up regulation of LT-α, TRAF2, and NIK.
