Background: Hepatocellular carcinoma(HCC) is a common malignant tumor in the world, especially in China. As a member of the inhibitor of differentiation(Id) family, Id4 has been reported to function in many cancer typ...Background: Hepatocellular carcinoma(HCC) is a common malignant tumor in the world, especially in China. As a member of the inhibitor of differentiation(Id) family, Id4 has been reported to function in many cancer types, but relatively little is known about its role in HCC. The purpose of this study was to investigate the potential relationship between Id4 and HCC development and the underlying mechanism involving the function of Id4 in HCC.Methods: We used quantitative real?time polymerase chain reaction and Western blotting to examine the RNA and protein expression of Id4. In addition, we used Cell Counting Kit?8 assay and colony formation assay to identify the function of Id4 in the regulation of cell proliferation in human HCC.Results: We found that the expression of Id4 protein was up?regulated in tumor tissues from HCC patients. Over?expression of Id4 promoted HCC cell proliferation, clonogenicity in vitro, and tumorigenicity in vivo. Id4 knockdown experiments showed that silencing Id4 blocked the proliferation and colony formation ability of HCC cells in vitro. Furthermore, overexpression of CCAAT/enhancer?binding protein β inhibited Id4 expression in HCC cells.Conclusion: Id4 may be developed as a potent therapeutic agent for the treatment of HCC, but more details about the underlying mechanisms of action are needed.展开更多
Objective： The aim of the study was to construct a stable strain of recombined attenuated Salmonella typhimurium expressing NK4 gene, and observe the effect of the strain on the metastatic potentiality of HepG2 cells...Objective： The aim of the study was to construct a stable strain of recombined attenuated Salmonella typhimurium expressing NK4 gene, and observe the effect of the strain on the metastatic potentiality of HepG2 cells. Methods： The NK4 cDNA was isolated from PCAGGS/hNK4 plasmid by PCR, and subcloned into eukaryotic expression vector pcDNA4. The recombinant plasmid was electro-transferred into attenuated Salmonella typhimurium Ty21a to obtain the recombinant strain encoding NK4 gene （TPN）. Simultaneously, the recombinant attenuated Salmonella typhimurium carrying GFP gene （TPG） was also constructed. After the TPG and TPN were transferred into HepG2 cells, the transfection rate and the expression level of NK4 protein were detected by flow cytometry and ELISA, and the effects of expression product on the proliferation and migration of HepG2 and angiogenesis were observed. Results： The TPN and TPG were successfully constructed. Fortyeight hours after transfection with TPG, the infection rate was 82.58% ± 1.74%, and the expression level of NK4 protein in supernatant was （181.5 ± 11.7） ng/6 × 10^5 cells. The supematant had obviously depressant effect on the proliferative activity of HepG2 cells （P 〈 0.05）, and could obviously restrain the hepatocyte growth factor-mediated migration of tumor cells （P 〈 0.01）. The inhibitory effect of the expression product on the tumor angiopoiesis was obviously observed （P 〈 0.05）, without a dosage-effect relation. Conclusion： The TPN could effectively transfer tumor cells in vitro and express interest NK4 protein. The expression product could effectively inhibit the proliferation and migration of hepatocellular carcinoma cells and the tumor angiopoiesis.展开更多
KLFs家族是一类具有锌指结构的转录因子,参与多个生理活动的调控,在细胞增殖、凋亡、分化以及胚胎发育等过程中扮演了关键的角色,与真核基因转录调控密切相关.Kruppel样因子4(Kruppel like factor 4,KLF4)作为KLFs家族中重要的一员,KLF...KLFs家族是一类具有锌指结构的转录因子,参与多个生理活动的调控,在细胞增殖、凋亡、分化以及胚胎发育等过程中扮演了关键的角色,与真核基因转录调控密切相关.Kruppel样因子4(Kruppel like factor 4,KLF4)作为KLFs家族中重要的一员,KLF4在原发性肝癌组织中呈现高表达,但是KLF4在原发性肝细胞癌的作用机制仍需要进一步的探讨,本文就KLF4在原发性肝细胞癌中的生物学功能进行综述.展开更多
目的:探讨溴结构域蛋白4(bromodomaincontaining protein 4,BRD4)抑制剂JQ1对两种不同肝癌细胞系增殖、凋亡的影响.方法:用B R D4抑制剂J Q1处理H e p G2,Bel-7402细胞株,磺酰罗丹明B(sulforhodamine B,SRB)染色检测细胞活力;Ed U(5-eth...目的:探讨溴结构域蛋白4(bromodomaincontaining protein 4,BRD4)抑制剂JQ1对两种不同肝癌细胞系增殖、凋亡的影响.方法:用B R D4抑制剂J Q1处理H e p G2,Bel-7402细胞株,磺酰罗丹明B(sulforhodamine B,SRB)染色检测细胞活力;Ed U(5-ethynyl-2'-deoxyuridine)嵌入和Hoechst 33342染色法检测细胞增殖;以流式细胞术和Annexin V-FITC/PI双染法分别分析细胞周期、细胞凋亡;最后用Western blot法观察JQ1介导抗增殖作用主要下调蛋白C-myc在蛋白水平上的变化,进一步验证JQ1抑制肝癌细胞增殖的作用.同时探讨JQ1与目前肝癌细胞治疗药物甲苯磺酸索拉非尼片对肝癌细胞的比较和联合作用.结果:BRD4抑制剂JQ1可以以剂量依赖的方式显著抑制HCC细胞系:Hep G2细胞和B e l-7402细胞的活力,具体表现为:抑制其细胞增殖;Annexin V-FITC/PI双染法分析发现能促进其凋亡;同时C-myc蛋白表达的也被显著抑制.与JQ1、索拉非尼片单药应用相比,小剂量的JQ1与索拉非尼片联合应用,HCC细胞的生长抑制率增加,凋亡率也增加,提示BRD4抑制剂JQ1联合索拉非尼片对细胞生长的抑制作用要强于JQ1或索拉非尼片单独用药.结论:BRD4抑制剂JQ1可能是一种潜在的可治疗肝细胞癌的新型药物.展开更多
基金supported in part by grants from the National Key Program for Basic Research of China(973)(No.2015CB553905)the National Natural Science Foundation of China(Nos.81272438,81472726,81301859)+1 种基金the Key Discipline and Specialty Foundation of Shanghai Municipal Commission of Health and Family Planning,the National Key Sci-Tech Special Project of China(No.2013ZX10002–011)the SKLORG Research Foundation(Nos.91-12-04,91-13-02,91-14-09)
文摘Background: Hepatocellular carcinoma(HCC) is a common malignant tumor in the world, especially in China. As a member of the inhibitor of differentiation(Id) family, Id4 has been reported to function in many cancer types, but relatively little is known about its role in HCC. The purpose of this study was to investigate the potential relationship between Id4 and HCC development and the underlying mechanism involving the function of Id4 in HCC.Methods: We used quantitative real?time polymerase chain reaction and Western blotting to examine the RNA and protein expression of Id4. In addition, we used Cell Counting Kit?8 assay and colony formation assay to identify the function of Id4 in the regulation of cell proliferation in human HCC.Results: We found that the expression of Id4 protein was up?regulated in tumor tissues from HCC patients. Over?expression of Id4 promoted HCC cell proliferation, clonogenicity in vitro, and tumorigenicity in vivo. Id4 knockdown experiments showed that silencing Id4 blocked the proliferation and colony formation ability of HCC cells in vitro. Furthermore, overexpression of CCAAT/enhancer?binding protein β inhibited Id4 expression in HCC cells.Conclusion: Id4 may be developed as a potent therapeutic agent for the treatment of HCC, but more details about the underlying mechanisms of action are needed.
基金Supported by a grant from the National Natural Science Foundation of China (No.30772572)two research grants from Postdoctors in China (No.20060390192,200801243)+1 种基金a grant from the Natural Science Foundation of Gansu Province (No.0803RJZA061)a research grant for science from Gansu Provincial Sci. & Tech. Department (No.0708NKCA128)
文摘Objective： The aim of the study was to construct a stable strain of recombined attenuated Salmonella typhimurium expressing NK4 gene, and observe the effect of the strain on the metastatic potentiality of HepG2 cells. Methods： The NK4 cDNA was isolated from PCAGGS/hNK4 plasmid by PCR, and subcloned into eukaryotic expression vector pcDNA4. The recombinant plasmid was electro-transferred into attenuated Salmonella typhimurium Ty21a to obtain the recombinant strain encoding NK4 gene （TPN）. Simultaneously, the recombinant attenuated Salmonella typhimurium carrying GFP gene （TPG） was also constructed. After the TPG and TPN were transferred into HepG2 cells, the transfection rate and the expression level of NK4 protein were detected by flow cytometry and ELISA, and the effects of expression product on the proliferation and migration of HepG2 and angiogenesis were observed. Results： The TPN and TPG were successfully constructed. Fortyeight hours after transfection with TPG, the infection rate was 82.58% ± 1.74%, and the expression level of NK4 protein in supernatant was （181.5 ± 11.7） ng/6 × 10^5 cells. The supematant had obviously depressant effect on the proliferative activity of HepG2 cells （P 〈 0.05）, and could obviously restrain the hepatocyte growth factor-mediated migration of tumor cells （P 〈 0.01）. The inhibitory effect of the expression product on the tumor angiopoiesis was obviously observed （P 〈 0.05）, without a dosage-effect relation. Conclusion： The TPN could effectively transfer tumor cells in vitro and express interest NK4 protein. The expression product could effectively inhibit the proliferation and migration of hepatocellular carcinoma cells and the tumor angiopoiesis.
文摘KLFs家族是一类具有锌指结构的转录因子,参与多个生理活动的调控,在细胞增殖、凋亡、分化以及胚胎发育等过程中扮演了关键的角色,与真核基因转录调控密切相关.Kruppel样因子4(Kruppel like factor 4,KLF4)作为KLFs家族中重要的一员,KLF4在原发性肝癌组织中呈现高表达,但是KLF4在原发性肝细胞癌的作用机制仍需要进一步的探讨,本文就KLF4在原发性肝细胞癌中的生物学功能进行综述.
文摘目的:探讨溴结构域蛋白4(bromodomaincontaining protein 4,BRD4)抑制剂JQ1对两种不同肝癌细胞系增殖、凋亡的影响.方法:用B R D4抑制剂J Q1处理H e p G2,Bel-7402细胞株,磺酰罗丹明B(sulforhodamine B,SRB)染色检测细胞活力;Ed U(5-ethynyl-2'-deoxyuridine)嵌入和Hoechst 33342染色法检测细胞增殖;以流式细胞术和Annexin V-FITC/PI双染法分别分析细胞周期、细胞凋亡;最后用Western blot法观察JQ1介导抗增殖作用主要下调蛋白C-myc在蛋白水平上的变化,进一步验证JQ1抑制肝癌细胞增殖的作用.同时探讨JQ1与目前肝癌细胞治疗药物甲苯磺酸索拉非尼片对肝癌细胞的比较和联合作用.结果:BRD4抑制剂JQ1可以以剂量依赖的方式显著抑制HCC细胞系:Hep G2细胞和B e l-7402细胞的活力,具体表现为:抑制其细胞增殖;Annexin V-FITC/PI双染法分析发现能促进其凋亡;同时C-myc蛋白表达的也被显著抑制.与JQ1、索拉非尼片单药应用相比,小剂量的JQ1与索拉非尼片联合应用,HCC细胞的生长抑制率增加,凋亡率也增加,提示BRD4抑制剂JQ1联合索拉非尼片对细胞生长的抑制作用要强于JQ1或索拉非尼片单独用药.结论:BRD4抑制剂JQ1可能是一种潜在的可治疗肝细胞癌的新型药物.
