目的:检测肝细胞肝癌(hepatocellular carcinoma,HCC)癌组织、癌旁组织(adjacent normal liver tissue,ANLT)及不同侵袭能力细胞系中极性蛋白AF-6 m RNA的表达情况,分析其在不同组织及不同细胞系中的表达差异及意义.方法:利用实时定量PC...目的:检测肝细胞肝癌(hepatocellular carcinoma,HCC)癌组织、癌旁组织(adjacent normal liver tissue,ANLT)及不同侵袭能力细胞系中极性蛋白AF-6 m RNA的表达情况,分析其在不同组织及不同细胞系中的表达差异及意义.方法:利用实时定量PCR检测(real-time quantitative,q RT-PCR)检测AF-6 m RNA在3 0对癌组织、A N LT和4种细胞系(L 0 2、Hep G2、MHCC97-H和HCCLM3)中的表达情况.结果:AF-6 m RNA在93.3%(28/30)的HCC中呈低表达;正常肝细胞株L02中AF-6 m RNA含量明显高于肝癌细胞株(P<0.05);高侵袭转移能力的细胞系MHCC97-H及HCCLM3只有极低的AF-6 m RNA表达,且明显低于低侵袭转移能力的细胞系Hep G2(P<0.05).结论:AF-6 m RNA在肝癌中的低表达可能与高侵袭能力相关,提示AF-6 m RNA在将来可能会成为治疗侵袭性HCC的潜在靶点.展开更多
The partitioning-defective 3 （Par3）, a key component in the conserved Par3/Par6/aPKC complex, plays fundamental roles in cell polarity. Herein we report the identification of Ku70 and Ku80 as novel Par3-interacting ...The partitioning-defective 3 （Par3）, a key component in the conserved Par3/Par6/aPKC complex, plays fundamental roles in cell polarity. Herein we report the identification of Ku70 and Ku80 as novel Par3-interacting proteins through an in vitro binding assay followed by liquid chromatography-tandem mass spectrometry. Ku70/Ku80 proteins are two key regulatory subunits of the DNA-dependent protein kinase （DNA-PK）, which plays an essential role in repairing double-strand DNA breaks （DSBs）. We determined that the nuclear association of Par3 with Ku70/Ku80 was enhanced by γ-irradiation （IR）, a potent DSB inducer. Furthermore, DNA-PKcs, the catalytic subunit of DNA-PK, interacted with the Par3/Ku70/Ku80 complex in response to IR. Par3 over-expression or knockdown was capable of up- or downregulating DNA-PK activity, respectively. Moreover, the Par3 knockdown cells were found to be defective in random plasmid integration, defective in DSB repair following IR, and radiosensitive, phenotypes similar to that of Ku70 knockdown cells. These findings identify Par3 as a novel component of the DNA-PK complex and implicate an unexpected link of cell polarity to DSB repair.展开更多
The author affiliations were mixed up in the previous published version. The third fund number of National NaturalScience Foundation of China in the Acknowledgments was wrong, it should be "30270335". The Sh...The author affiliations were mixed up in the previous published version. The third fund number of National NaturalScience Foundation of China in the Acknowledgments was wrong, it should be "30270335". The Shanghai MunicipalCouncil for Science and Technology (No.06DZ22032) was missed in the Acknowledgments. There are some labelingand production errors in Figure 2A, Figure 3B and 3C, Figure 5C, Figure 6B and 6E, Figure 7B and 7D. In Figure 2A,left panel "A431" should be "Par3". In Figure 3B and 3C, "anti-Par3CT" should be "anti-Par3LCT", "GST-Par3CT"should be "GST-Par3LCT". In Figure 5C, the second arrow indicating "Lamin B" should be "[3-tubulin". In Figure 6Bright panel, the molecular weight for ?-actin should be "43" instead of"200". In Figure 6E, "Par3" should be "Par3i". Themolecular weight for the DNA-PKcs panel should be the same as the p-DNA-PKcs. In Figure 7B, the time point "240"in the left panel should be "120"; in the right panel of Figure 7B, the title for the y axis should be "DNA released (%)".In Figure 7D, the title for the y axis should be "Survival (%)", and the scale for the y axis should be "100, 10 and 1". These corrections do not affect the conclusions of the study. We apologize for any inconvenience this may havecaused.展开更多
目的探讨极性蛋白Par3(partitioning defective protein 3,Par3)和磷酸化的信号传导子和转录激活子3(p-signal transducers and activators of transcription 3,p-STAT3)在宫颈病变组织中的表达及临床意义。方法采用免疫组织化学染色法...目的探讨极性蛋白Par3(partitioning defective protein 3,Par3)和磷酸化的信号传导子和转录激活子3(p-signal transducers and activators of transcription 3,p-STAT3)在宫颈病变组织中的表达及临床意义。方法采用免疫组织化学染色法检测Par3和p-STAT3蛋白在慢性宫颈炎、宫颈上皮内瘤变Ⅱ-Ⅲ(cervical intraepithelial neoplasia Ⅱ-Ⅲ,CIN Ⅱ-Ⅲ)以及宫颈鳞状细胞癌(cervical squamous carcinoma,CSCC)和淋巴结转移灶中的表达情况,分析其与临床病理参数之间的关系。结果Par3蛋白在慢性宫颈炎、CIN Ⅱ-Ⅲ、CSCC和淋巴结转移灶中的蛋白表达量呈现逐渐降低趋势(P<0.001);上述4种组织中p-STAT3蛋白的表达量是随着宫颈病变加重而逐渐增高(P=0.0062)。比较二者与临床病理参数的关系发现,宫颈鳞癌中Par3表达与肿瘤淋巴结转移(P=0.003)以及组织学分化程度(P=0.017)有关,宫颈鳞癌组织p-STAT3阳性表达与肿瘤临床分期、组织学分化程度有关(P=0.001)。CINⅡ-Ⅲ组织中极性蛋白Par3与p-STAT3蛋白水平呈负相关(r=-5.553,P<0.05)。结论宫颈鳞癌中极性蛋白Par3低表达,p-STAT3蛋白高表达,二者表达差异与宫颈鳞癌发生、组织学分化、肿瘤淋巴结转移以及临床分期有关。展开更多
文摘目的:检测肝细胞肝癌(hepatocellular carcinoma,HCC)癌组织、癌旁组织(adjacent normal liver tissue,ANLT)及不同侵袭能力细胞系中极性蛋白AF-6 m RNA的表达情况,分析其在不同组织及不同细胞系中的表达差异及意义.方法:利用实时定量PCR检测(real-time quantitative,q RT-PCR)检测AF-6 m RNA在3 0对癌组织、A N LT和4种细胞系(L 0 2、Hep G2、MHCC97-H和HCCLM3)中的表达情况.结果:AF-6 m RNA在93.3%(28/30)的HCC中呈低表达;正常肝细胞株L02中AF-6 m RNA含量明显高于肝癌细胞株(P<0.05);高侵袭转移能力的细胞系MHCC97-H及HCCLM3只有极低的AF-6 m RNA表达,且明显低于低侵袭转移能力的细胞系Hep G2(P<0.05).结论:AF-6 m RNA在肝癌中的低表达可能与高侵袭能力相关,提示AF-6 m RNA在将来可能会成为治疗侵袭性HCC的潜在靶点.
基金This work was supported by the grants from National Natural Science Foundation of China(Nos.30170208,30623003 and 30170208)from the Ministry of Science and Technology,China(No.2001AA233031 and No.2001CB510205)from US NIH(CA50519)(to DJ Chen).
文摘The partitioning-defective 3 （Par3）, a key component in the conserved Par3/Par6/aPKC complex, plays fundamental roles in cell polarity. Herein we report the identification of Ku70 and Ku80 as novel Par3-interacting proteins through an in vitro binding assay followed by liquid chromatography-tandem mass spectrometry. Ku70/Ku80 proteins are two key regulatory subunits of the DNA-dependent protein kinase （DNA-PK）, which plays an essential role in repairing double-strand DNA breaks （DSBs）. We determined that the nuclear association of Par3 with Ku70/Ku80 was enhanced by γ-irradiation （IR）, a potent DSB inducer. Furthermore, DNA-PKcs, the catalytic subunit of DNA-PK, interacted with the Par3/Ku70/Ku80 complex in response to IR. Par3 over-expression or knockdown was capable of up- or downregulating DNA-PK activity, respectively. Moreover, the Par3 knockdown cells were found to be defective in random plasmid integration, defective in DSB repair following IR, and radiosensitive, phenotypes similar to that of Ku70 knockdown cells. These findings identify Par3 as a novel component of the DNA-PK complex and implicate an unexpected link of cell polarity to DSB repair.
基金supported by the grants from National Natural Science Foundation of China(No.30170208,30623003 and 30270335)from the Ministry of Science and Technology,China(No.2001AA233031 and No.2001CB510205)+1 种基金from the Shanghai Municipal Council for Science and Technology(No.06DZ22032)from US NIH(CA50519)(to DJ Chen).
文摘The author affiliations were mixed up in the previous published version. The third fund number of National NaturalScience Foundation of China in the Acknowledgments was wrong, it should be "30270335". The Shanghai MunicipalCouncil for Science and Technology (No.06DZ22032) was missed in the Acknowledgments. There are some labelingand production errors in Figure 2A, Figure 3B and 3C, Figure 5C, Figure 6B and 6E, Figure 7B and 7D. In Figure 2A,left panel "A431" should be "Par3". In Figure 3B and 3C, "anti-Par3CT" should be "anti-Par3LCT", "GST-Par3CT"should be "GST-Par3LCT". In Figure 5C, the second arrow indicating "Lamin B" should be "[3-tubulin". In Figure 6Bright panel, the molecular weight for ?-actin should be "43" instead of"200". In Figure 6E, "Par3" should be "Par3i". Themolecular weight for the DNA-PKcs panel should be the same as the p-DNA-PKcs. In Figure 7B, the time point "240"in the left panel should be "120"; in the right panel of Figure 7B, the title for the y axis should be "DNA released (%)".In Figure 7D, the title for the y axis should be "Survival (%)", and the scale for the y axis should be "100, 10 and 1". These corrections do not affect the conclusions of the study. We apologize for any inconvenience this may havecaused.
