Objective Cytochrome P450 2E1 （CYP2E1） is an important metabolizing enzyme involved in oxidative stress responses to benzene, a chemical associated with bone marrow toxicity and leukemia, We aimed to identify the CY...Objective Cytochrome P450 2E1 （CYP2E1） is an important metabolizing enzyme involved in oxidative stress responses to benzene, a chemical associated with bone marrow toxicity and leukemia, We aimed to identify the CYP2E1 genetic biomarkers of susceptibility to benzene toxicity in support of environmental and occupational exposure prevention, and to test whether a model using immortal human lymphocytes might be an efficient tool for detecting genetic biomarkers. Methods Immortalized human lymphocyte cell lines with independent genotypes on four CYP2E1 SNP sites were induced with 0.01% phenol, a metabolite of benzene. CYP2E1 gene function was evaluated by mRNA expression and enzyme activity. DNA damage was measured by Single-Cell Gel Electrophoresis （SCGE）. Results Among the four SNPs, cells with rs2070673TT and rs2030920CC showed higher levels of ~YP2E1 transcription and enzymatic activity than the other genotypes in the same SNP site. Cells with higher gene expression genotypes also showed higher comet rates compared with lower gene expression genotypes. Conclusion These results suggest that CYP2E1 rs2070673 and rs2030920 might be the genetic biomarkers of susceptibility to benzene toxicity and that the immortalized human lymphocytes model might be an efficient tool for the detection of genetic biomarkers of susceptibility to chemicals.展开更多
Objective To investigate the role of TNF receptor-associated factor 2 (TRAF-2) and TRAF6 in CD40-induced nuclear factor-κB (NF-κB) signaling pathway and whether CD40 signaling requires TRAF2. Methods Human B cell li...Objective To investigate the role of TNF receptor-associated factor 2 (TRAF-2) and TRAF6 in CD40-induced nuclear factor-κB (NF-κB) signaling pathway and whether CD40 signaling requires TRAF2. Methods Human B cell lines were transfected with plasmids expressing wild type TRAF2 or dominant negative TRAF2,TRAF2-shRNA,or TRAF6-shRNA. The activation of NF-κB was detected by Western blot,kinase assay,transfactor enzyme-linked immunosorbent assay (ELISA),and fluorescence resonance energy transfer (FRET). Analysis of the role of TRAF-2 and TRAF-6 in CD40-mediated NF-κB activity was examined following stimulation with recombinant CD154. Results TRAF2 induced activity of IκB-kinases (IKKα,IKKi/ε),phosphorylation of IκBα,as well as nuclear translocation and phosphorylation of p65/RelA. In contrast,TRAF6 strongly induced NF-κB activation and nuclear translocation of p65 as well as p50 and c-Rel. Engagement of CD154-induced nuclear translocation of p65 was inhibited by a TRAF6-shRNA,but conversely was enhanced by a TRAF2-shRNA. Examination of direct interactions between CD40 and TRAFs by FRET documented that both TRAF2 and TRAF6 directly interacted with CD40. However,the two TRAFs competed for CD40 binding. Conclusions These results indicate that TRAF2 can signal in human B cells,but it is not essential for CD40-mediated NF-κB activation. Moreover,TRAF2 can compete with TRAF6 for CD40 binding,and thereby limit the capacity of CD40 engagement to induce NF-κB activation.展开更多
目的:探讨合并乙型肝炎病毒(hepatitis B virus,HBV)感染对弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)患者血清细胞因子表达水平的影响及其临床意义。方法:选择2013年9月至2014年4月于复旦大学附属中山医院血液科住院接...目的:探讨合并乙型肝炎病毒(hepatitis B virus,HBV)感染对弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)患者血清细胞因子表达水平的影响及其临床意义。方法:选择2013年9月至2014年4月于复旦大学附属中山医院血液科住院接受治疗的DLBCL患者共77例,测定初治及完全缓解后DLBCL患者血清中肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)、可溶性白细胞介素2受体(soluble interleukin-2receptor,sIL-2r)、白细胞介素-6(interleukin-6,IL-6)及白细胞介素-10(IL-10)的表达水平;以20例年龄匹配的同期健康体检者作为对照。进一步分析乙型肝炎病毒表面抗原(HBsAg)阳性及阴性DLBCL患者血清中细胞因子水平的差异。结果:初治DLBCL患者血清TNF-α、sIL-2r及IL-6水平均明显高于健康对照组(均P<0.05);缓解期DLBCL患者血清TNF-α和sIL-2r水平明显低于初治患者(均P<0.05),但仍明显高于健康对照组(均P<0.05)。初治DLBCL患者中,HBsAg阳性者的上述血清细胞因子水平与HBsAg阴性者差异无统计学意义;缓解期DLBCL患者中,HBsAg阳性者的血清sIL-2r水平显著高于HBsAg阴性者(P<0.05)。结论:DLBCL患者血清TNF-α、sIL-2r和IL-6水平升高,其中TNF-α及sIL-2r水平与疾病缓解有关。HBsAg阳性DLBCL患者在疾病缓解后的sIL-2r水平明显高于HBsAg阴性者,sIL-2r水平与HBsAg阳性DLBCL患者预后不良之间的相关性还有待进一步研究。展开更多
基金supported by the National Natural Science Foundation of China (Grant No: 30671731, 30901168)the Doctoral Program of Higher Education of China (Grant No: 20070286069)
文摘Objective Cytochrome P450 2E1 （CYP2E1） is an important metabolizing enzyme involved in oxidative stress responses to benzene, a chemical associated with bone marrow toxicity and leukemia, We aimed to identify the CYP2E1 genetic biomarkers of susceptibility to benzene toxicity in support of environmental and occupational exposure prevention, and to test whether a model using immortal human lymphocytes might be an efficient tool for detecting genetic biomarkers. Methods Immortalized human lymphocyte cell lines with independent genotypes on four CYP2E1 SNP sites were induced with 0.01% phenol, a metabolite of benzene. CYP2E1 gene function was evaluated by mRNA expression and enzyme activity. DNA damage was measured by Single-Cell Gel Electrophoresis （SCGE）. Results Among the four SNPs, cells with rs2070673TT and rs2030920CC showed higher levels of ~YP2E1 transcription and enzymatic activity than the other genotypes in the same SNP site. Cells with higher gene expression genotypes also showed higher comet rates compared with lower gene expression genotypes. Conclusion These results suggest that CYP2E1 rs2070673 and rs2030920 might be the genetic biomarkers of susceptibility to benzene toxicity and that the immortalized human lymphocytes model might be an efficient tool for the detection of genetic biomarkers of susceptibility to chemicals.
基金Supported by Key Projects of the National Science & Technology Pillar Program in the Eleventh Five-year Plan Period (2008-BAI59B02)
文摘Objective To investigate the role of TNF receptor-associated factor 2 (TRAF-2) and TRAF6 in CD40-induced nuclear factor-κB (NF-κB) signaling pathway and whether CD40 signaling requires TRAF2. Methods Human B cell lines were transfected with plasmids expressing wild type TRAF2 or dominant negative TRAF2,TRAF2-shRNA,or TRAF6-shRNA. The activation of NF-κB was detected by Western blot,kinase assay,transfactor enzyme-linked immunosorbent assay (ELISA),and fluorescence resonance energy transfer (FRET). Analysis of the role of TRAF-2 and TRAF-6 in CD40-mediated NF-κB activity was examined following stimulation with recombinant CD154. Results TRAF2 induced activity of IκB-kinases (IKKα,IKKi/ε),phosphorylation of IκBα,as well as nuclear translocation and phosphorylation of p65/RelA. In contrast,TRAF6 strongly induced NF-κB activation and nuclear translocation of p65 as well as p50 and c-Rel. Engagement of CD154-induced nuclear translocation of p65 was inhibited by a TRAF6-shRNA,but conversely was enhanced by a TRAF2-shRNA. Examination of direct interactions between CD40 and TRAFs by FRET documented that both TRAF2 and TRAF6 directly interacted with CD40. However,the two TRAFs competed for CD40 binding. Conclusions These results indicate that TRAF2 can signal in human B cells,but it is not essential for CD40-mediated NF-κB activation. Moreover,TRAF2 can compete with TRAF6 for CD40 binding,and thereby limit the capacity of CD40 engagement to induce NF-κB activation.
文摘目的:探讨合并乙型肝炎病毒(hepatitis B virus,HBV)感染对弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)患者血清细胞因子表达水平的影响及其临床意义。方法:选择2013年9月至2014年4月于复旦大学附属中山医院血液科住院接受治疗的DLBCL患者共77例,测定初治及完全缓解后DLBCL患者血清中肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)、可溶性白细胞介素2受体(soluble interleukin-2receptor,sIL-2r)、白细胞介素-6(interleukin-6,IL-6)及白细胞介素-10(IL-10)的表达水平;以20例年龄匹配的同期健康体检者作为对照。进一步分析乙型肝炎病毒表面抗原(HBsAg)阳性及阴性DLBCL患者血清中细胞因子水平的差异。结果:初治DLBCL患者血清TNF-α、sIL-2r及IL-6水平均明显高于健康对照组(均P<0.05);缓解期DLBCL患者血清TNF-α和sIL-2r水平明显低于初治患者(均P<0.05),但仍明显高于健康对照组(均P<0.05)。初治DLBCL患者中,HBsAg阳性者的上述血清细胞因子水平与HBsAg阴性者差异无统计学意义;缓解期DLBCL患者中,HBsAg阳性者的血清sIL-2r水平显著高于HBsAg阴性者(P<0.05)。结论:DLBCL患者血清TNF-α、sIL-2r和IL-6水平升高,其中TNF-α及sIL-2r水平与疾病缓解有关。HBsAg阳性DLBCL患者在疾病缓解后的sIL-2r水平明显高于HBsAg阴性者,sIL-2r水平与HBsAg阳性DLBCL患者预后不良之间的相关性还有待进一步研究。
