p28, a 28kD protein from toad (Bufo bufo gargarizans) oocytes, was identified by using p13sucl-agarose affinity chromatography. Sequence homology analysis of the full-length cDNA of p28 (Gene Bank accession number: AF...p28, a 28kD protein from toad (Bufo bufo gargarizans) oocytes, was identified by using p13sucl-agarose affinity chromatography. Sequence homology analysis of the full-length cDNA of p28 (Gene Bank accession number: AF 314091) indicated that it encodes a protein containing 224 amino-acids with about 55% identities and more than 70% positives to human, rat or mouse UCH-L1, and contains homological functional domains of UCH family. Anti-p28 monoclonal antibody, on injecting into the oocytes, could inhibit the progesterone-induced resumption of meiotic division in a dose-dependent manner. The recombinant protein p28 showed similar SDS/PAGE behaviors to the native one, and promoted ubiquitin ethyl ester hydrolysis, a classical catalytic reaction for ubiquitin carboxyl terminal hydrolases (UCHs). The results in this paper reveal that a novel protein, p28, exists in the toad oocytes, is a UCH L1 homolog, was engaged in the process of progesterone-induced oocyte maturation possibly through an involvement in protein turnover and degradation.展开更多
P28，a 28kD protein from toad (Bufo bufo gargarizans)oocytes,was identified by using P13^suc1-agarose affinity chromatography.Sequence homology analysis of the full-length cDNA of P28(Gene Bank accession number:AF 314...P28，a 28kD protein from toad (Bufo bufo gargarizans)oocytes,was identified by using P13^suc1-agarose affinity chromatography.Sequence homology analysis of the full-length cDNA of P28(Gene Bank accession number:AF 314091)indicated that it encodes a protein containing 224 amino-acids with about 55% iden-tities and more than 70%positives to encodes a protein containing 224 amino-acids with about 55% iden-tities and more than 70% positives to human, rat or mouse UCH-L1,and contains homological functional domains of UCH family.Anti-p28 monoclonal antibody,on injecting into the oocytes,could inhibit the progesterone-induced resumption of meiotic division in a dose-dependent manner.The recombinant protein P28 showed similar SDS/PAGE behaviors to the native one,and promoted ubiquitin ethyl ester hydrolysis,a classical catalytic reaction for ubiquitin carboxyl terminal hydrolases(UCHs).The results in this paper reveal that a novel protein,p28 ,exists in the toad oocytes,is a UCH Ll homolog,was engaged in the process of progesterone-induced oocyte maturation possibly through an involvement in protein turnover and degradation.展开更多
AIM: To investigate the expression of p28/gankyringene and its role in the carcinogenetic process of humanhepatocellular carcinoma (HCC).METHODS: 64 specimens of HCC and para-carcinomatissues, 22 specimens of non-tumo...AIM: To investigate the expression of p28/gankyringene and its role in the carcinogenetic process of humanhepatocellular carcinoma (HCC).METHODS: 64 specimens of HCC and para-carcinomatissues, 22 specimens of non-tumor liver tissues (7normal, 15 cirrhosis), 10 specimens of normal humantissues and 5 hepatoma cell lines were studied for theexpression of p28/gankyrin by Northern blot. Theexpression of p28/gankyrin protein was detectedimmunohistochemically by using the specificpolyclonal antibody. RESULTS: Northern blot analysis indicated that theexpression of p28/gankyrin mRNA was intensivelydistributed in brain and heart, weakly in lung, spleenand muscle, undetectable in digestive system includingliver, pancreas, stomach, small and large intestines.p28/gankyrin mRNA was absent in normal liver, weaklydetected in liver cirrhosis and in 18 of 64 para-carcinoma liver tissues. In contrast, the expression ofp28/gankyrin mRNA was intensively detected in ali 5hepatoma cell lines tested, markedly increased in 57of 64 and moderately increased in 5 of 64 HCC samples.In comparison with liver cirrhosis and para-carcinomaliver tissues, the average expression of p28/gankyrin mRNA in HCC was increased 3.6- (2.901+0.507 vs 0.805 + 0.252, P＜0.05) and 5.2-fold (2.901 +0.507 vs 0.557+0.203, p＜0.01), respectively, Inaddition, p28/gankyrin mRNA expression level washigher in HCC with portal vein tumor thrombus andmicroscopic hepatic vein involvement (P--0.021 andP=-0.047, respectively). The overexpression of p28/gankyrin protein in HCC was targeted in hepatic tumorcells, not in bile duct cells and other interstitial cells.CONCLUSION: Overexpression of p28/gankyrin in HCCplays an important role and contributes to themetastasis potential in the process of carcinogenesis.p28/gankyrin may become a specific biological tissuemarker for the pathological diagnosis of HCC.展开更多
It has been shown that oncoprotein p28GANK, which is consistently overexpressed in human hepatocellular carcinoma （HCC）, plays a critical role in tumorigenesis of HCC. However, the underlying mechanism remains uncle...It has been shown that oncoprotein p28GANK, which is consistently overexpressed in human hepatocellular carcinoma （HCC）, plays a critical role in tumorigenesis of HCC. However, the underlying mechanism remains unclear. Here, we demonstrated that p28GANK inhibits apoptosis in HCC cells induced by the endoplasmic reticulum （ER） stress. During ER stress, p28GANK enhances the unfolded protein response, promotes ER recovery from translational repression, and thereby facilitates cell＇s ability to cope with the stress conditions. Furthermore, p28GANK upregulates glucose-regulated protein 78 （GRP78）, a key ER chaperone protein, which subsequently enhances the ER folding capacity and promotes recovery from ER stress. We also demonstrated that p28GANK increases p38 mitogen-activated protein kinase and Akt phosphorylation, and inhibits nuclear factor kappa B （NF-κB） activation under ER stress, which in turn contributes to GRP78 upregulation. Taken together, our results indicate that p28GANK inhibits ER stress-induced apoptosis in HCC cells, at least in part, by enhancing the adaptive response and GRP78 expression. We propose that p28GANK has potential implications for HCC progression under the ER stress conditions.展开更多
AIM: To observe the gene and protein expression changes of p28GANK in regenerating liver tissues, and to reveal the biological function of p28GANK on the regulation of liver regeneration.METHODS: One hundred and thirt...AIM: To observe the gene and protein expression changes of p28GANK in regenerating liver tissues, and to reveal the biological function of p28GANK on the regulation of liver regeneration.METHODS: One hundred and thirty two adult male Sprague-Dawley rats were selected, weighing 200-250 g,and divided randomly into sham operation (SO) group and partial hepatectomy (PH) group. Each group had eleven time points: 0, 2, 6, 12, 24, 30, 48, 72, 120, 168 and 240 h,six rats were in each time point. The rats were undergone 70 % PH under methoxyflurane anesthesia by resection of the anterior and left lateral lobes of the liver. SO was conducted by laparotomy plus slight mobilization of the liver without resection. Liver specimens were collected at the indicated time points after PH or SO. The expression level of p28GANK mRNA was determined by Northern blot as well as at protein level via immunohistochemical staining.The expressions of p28GANK mRNA in these tissues were analyzed by imaging analysis system of FLA-2000 FUJIFILM and one way analysis of variance. The protein expressions of p28GANK in these tissues were analyzed with Fromowitz'method and Rank sum test.RESULTS: The expression of p28GANK mRNA in bhe regenerating liver tissues possessed two transcripts, which were 1.5 kb and 1.0 kb. There was a significantly different expression patterns of p28GANK mRNA between SO and PH groups (P<0.01). The expression of p28GANK mRNA increased 2 h after PH, the peak time was 72 h (SO group: 163.83±1.4720; PH group: 510.5±17.0499, P<0.01). There was a significant difference in the 1.5 kb transcript, which decreased gradually after 72 hours. The protein expression of p28GANK was mainly in the cytoplasm of regenerating hepatocytes, and increased near the central region 24 h after PH, and became strongly positive at 48 h (+++, vs the other time points P<0.05),but decreased 72 h after PH.CONCLUSION: The expression of p28GANK mRNA increases in the early stage of rat liver regeneration, the protein expression of p28GANK is mainly in the c展开更多
p28^GANK （also known as PSMD 10, p28 and gankyrin） is an ankyrin repeat anti-apoptotic oncoprotein that is commonly overexpressed in hepatocellular carcinomas and increases the degradation of p53 and Rb. NF-IκB （n...p28^GANK （also known as PSMD 10, p28 and gankyrin） is an ankyrin repeat anti-apoptotic oncoprotein that is commonly overexpressed in hepatocellular carcinomas and increases the degradation of p53 and Rb. NF-IκB （nuclear factor-κB） is known to be sequestered in the cytoplasm by IκB （inhibitor of NF-κB） proteins [1, 2], but much less is known about the cytoplasmic retention of NF-κB by other cellular proteins. Here we show that p28^GANK inhibits NF-κB activity. As a nuclear-cytoplasmic shuttling protein, p28^GANK directly binds to NF-κB/RelA and exports RelA from nucleus through a chromosomal region maintenance-1 （CRM-1） dependent pathway, which results in the cytoplasmic retention of NF- κB/RelA. We demonstrate that all the ankyrin repeats of p28^GANK are required for the interaction with RelA and that the N terminus of p28^GANK, which contains the nuclear export sequence （NES）, is responsible for suppressing NF-κB/RelA nuclear translocation. These results suggest that overexpression of p28^GANK prevents the nuclear localization and inhibits the activity of NF-κB/RelA.展开更多
基金This work is supported by National Natural Sci-ence Fundation of China (Grant 39770370), and National Laboratory of Contraceptives and Devices Re-search affiliated with Shanghai lnstitute of Planned Parenthood Research.
文摘p28, a 28kD protein from toad (Bufo bufo gargarizans) oocytes, was identified by using p13sucl-agarose affinity chromatography. Sequence homology analysis of the full-length cDNA of p28 (Gene Bank accession number: AF 314091) indicated that it encodes a protein containing 224 amino-acids with about 55% identities and more than 70% positives to human, rat or mouse UCH-L1, and contains homological functional domains of UCH family. Anti-p28 monoclonal antibody, on injecting into the oocytes, could inhibit the progesterone-induced resumption of meiotic division in a dose-dependent manner. The recombinant protein p28 showed similar SDS/PAGE behaviors to the native one, and promoted ubiquitin ethyl ester hydrolysis, a classical catalytic reaction for ubiquitin carboxyl terminal hydrolases (UCHs). The results in this paper reveal that a novel protein, p28, exists in the toad oocytes, is a UCH L1 homolog, was engaged in the process of progesterone-induced oocyte maturation possibly through an involvement in protein turnover and degradation.
文摘白细胞介素-27(interleukin-27,IL-27)由p28和EB病毒诱导基因3(Epstein-Barr virus induced gene 3,EBB)组成,是IL-6/IL-12细胞因子家族中的成员之一,其受体由gp130和WSX1(也称为T细胞因子受体)组成.IL-27可作用于T细胞、B细胞、巨噬细胞、树突状细胞、自然杀伤细胞和非造血细胞等多种细胞类型.目前许多研究发现不同的T细胞亚群对IL-27有不同的应答.IL-27具有促炎和抗炎双重作用,主要通过Janus激酶(Janus kinase,JAK)/信号转导和转录激活剂(signal transducer and activator of transcription,STAT)通路和丝裂原活化蛋白激酶(mitogen activated protein kinase,MAPK)通路在T细胞不同亚群的免疫调节中发挥重要作用.
文摘P28，a 28kD protein from toad (Bufo bufo gargarizans)oocytes,was identified by using P13^suc1-agarose affinity chromatography.Sequence homology analysis of the full-length cDNA of P28(Gene Bank accession number:AF 314091)indicated that it encodes a protein containing 224 amino-acids with about 55% iden-tities and more than 70%positives to encodes a protein containing 224 amino-acids with about 55% iden-tities and more than 70% positives to human, rat or mouse UCH-L1,and contains homological functional domains of UCH family.Anti-p28 monoclonal antibody,on injecting into the oocytes,could inhibit the progesterone-induced resumption of meiotic division in a dose-dependent manner.The recombinant protein P28 showed similar SDS/PAGE behaviors to the native one,and promoted ubiquitin ethyl ester hydrolysis,a classical catalytic reaction for ubiquitin carboxyl terminal hydrolases(UCHs).The results in this paper reveal that a novel protein,p28 ,exists in the toad oocytes,is a UCH Ll homolog,was engaged in the process of progesterone-induced oocyte maturation possibly through an involvement in protein turnover and degradation.
基金the Chinese National Distinguished Young Scholar Awards,No.39825114Chinese National Key Project of Basic Research,No.G1998051210the Key Project of the Chinese National Natural Science Foundation,No.39830080.
文摘AIM: To investigate the expression of p28/gankyringene and its role in the carcinogenetic process of humanhepatocellular carcinoma (HCC).METHODS: 64 specimens of HCC and para-carcinomatissues, 22 specimens of non-tumor liver tissues (7normal, 15 cirrhosis), 10 specimens of normal humantissues and 5 hepatoma cell lines were studied for theexpression of p28/gankyrin by Northern blot. Theexpression of p28/gankyrin protein was detectedimmunohistochemically by using the specificpolyclonal antibody. RESULTS: Northern blot analysis indicated that theexpression of p28/gankyrin mRNA was intensivelydistributed in brain and heart, weakly in lung, spleenand muscle, undetectable in digestive system includingliver, pancreas, stomach, small and large intestines.p28/gankyrin mRNA was absent in normal liver, weaklydetected in liver cirrhosis and in 18 of 64 para-carcinoma liver tissues. In contrast, the expression ofp28/gankyrin mRNA was intensively detected in ali 5hepatoma cell lines tested, markedly increased in 57of 64 and moderately increased in 5 of 64 HCC samples.In comparison with liver cirrhosis and para-carcinomaliver tissues, the average expression of p28/gankyrin mRNA in HCC was increased 3.6- (2.901+0.507 vs 0.805 + 0.252, P＜0.05) and 5.2-fold (2.901 +0.507 vs 0.557+0.203, p＜0.01), respectively, Inaddition, p28/gankyrin mRNA expression level washigher in HCC with portal vein tumor thrombus andmicroscopic hepatic vein involvement (P--0.021 andP=-0.047, respectively). The overexpression of p28/gankyrin protein in HCC was targeted in hepatic tumorcells, not in bile duct cells and other interstitial cells.CONCLUSION: Overexpression of p28/gankyrin in HCCplays an important role and contributes to themetastasis potential in the process of carcinogenesis.p28/gankyrin may become a specific biological tissuemarker for the pathological diagnosis of HCC.
文摘It has been shown that oncoprotein p28GANK, which is consistently overexpressed in human hepatocellular carcinoma （HCC）, plays a critical role in tumorigenesis of HCC. However, the underlying mechanism remains unclear. Here, we demonstrated that p28GANK inhibits apoptosis in HCC cells induced by the endoplasmic reticulum （ER） stress. During ER stress, p28GANK enhances the unfolded protein response, promotes ER recovery from translational repression, and thereby facilitates cell＇s ability to cope with the stress conditions. Furthermore, p28GANK upregulates glucose-regulated protein 78 （GRP78）, a key ER chaperone protein, which subsequently enhances the ER folding capacity and promotes recovery from ER stress. We also demonstrated that p28GANK increases p38 mitogen-activated protein kinase and Akt phosphorylation, and inhibits nuclear factor kappa B （NF-κB） activation under ER stress, which in turn contributes to GRP78 upregulation. Taken together, our results indicate that p28GANK inhibits ER stress-induced apoptosis in HCC cells, at least in part, by enhancing the adaptive response and GRP78 expression. We propose that p28GANK has potential implications for HCC progression under the ER stress conditions.
文摘AIM: To observe the gene and protein expression changes of p28GANK in regenerating liver tissues, and to reveal the biological function of p28GANK on the regulation of liver regeneration.METHODS: One hundred and thirty two adult male Sprague-Dawley rats were selected, weighing 200-250 g,and divided randomly into sham operation (SO) group and partial hepatectomy (PH) group. Each group had eleven time points: 0, 2, 6, 12, 24, 30, 48, 72, 120, 168 and 240 h,six rats were in each time point. The rats were undergone 70 % PH under methoxyflurane anesthesia by resection of the anterior and left lateral lobes of the liver. SO was conducted by laparotomy plus slight mobilization of the liver without resection. Liver specimens were collected at the indicated time points after PH or SO. The expression level of p28GANK mRNA was determined by Northern blot as well as at protein level via immunohistochemical staining.The expressions of p28GANK mRNA in these tissues were analyzed by imaging analysis system of FLA-2000 FUJIFILM and one way analysis of variance. The protein expressions of p28GANK in these tissues were analyzed with Fromowitz'method and Rank sum test.RESULTS: The expression of p28GANK mRNA in bhe regenerating liver tissues possessed two transcripts, which were 1.5 kb and 1.0 kb. There was a significantly different expression patterns of p28GANK mRNA between SO and PH groups (P<0.01). The expression of p28GANK mRNA increased 2 h after PH, the peak time was 72 h (SO group: 163.83±1.4720; PH group: 510.5±17.0499, P<0.01). There was a significant difference in the 1.5 kb transcript, which decreased gradually after 72 hours. The protein expression of p28GANK was mainly in the cytoplasm of regenerating hepatocytes, and increased near the central region 24 h after PH, and became strongly positive at 48 h (+++, vs the other time points P<0.05),but decreased 72 h after PH.CONCLUSION: The expression of p28GANK mRNA increases in the early stage of rat liver regeneration, the protein expression of p28GANK is mainly in the c
基金Acknowledgments We thank Dr IM Verma （UCSD, USA） and Dr WC Greene （UCSF, USA） for the RelA, p50 and IKBct plasmids. Research was supported by grants from National Natural Science Foundation of China （30530790, 30620130434, 30428006 and 30500275）.
文摘p28^GANK （also known as PSMD 10, p28 and gankyrin） is an ankyrin repeat anti-apoptotic oncoprotein that is commonly overexpressed in hepatocellular carcinomas and increases the degradation of p53 and Rb. NF-IκB （nuclear factor-κB） is known to be sequestered in the cytoplasm by IκB （inhibitor of NF-κB） proteins [1, 2], but much less is known about the cytoplasmic retention of NF-κB by other cellular proteins. Here we show that p28^GANK inhibits NF-κB activity. As a nuclear-cytoplasmic shuttling protein, p28^GANK directly binds to NF-κB/RelA and exports RelA from nucleus through a chromosomal region maintenance-1 （CRM-1） dependent pathway, which results in the cytoplasmic retention of NF- κB/RelA. We demonstrate that all the ankyrin repeats of p28^GANK are required for the interaction with RelA and that the N terminus of p28^GANK, which contains the nuclear export sequence （NES）, is responsible for suppressing NF-κB/RelA nuclear translocation. These results suggest that overexpression of p28^GANK prevents the nuclear localization and inhibits the activity of NF-κB/RelA.
