To investigate the roles of apoptosis and the Fas system in the process of liver cirrhosis converting into hepatocellular carcinoma , expression of Fas and Fas ligand in 49 LC and 36 HCC samples was detected by i...To investigate the roles of apoptosis and the Fas system in the process of liver cirrhosis converting into hepatocellular carcinoma , expression of Fas and Fas ligand in 49 LC and 36 HCC samples was detected by immunohistochemical method. Apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling method. Serum soluble Fas levels in 28 cases of LC and 27 cases of HCC were measured by enzyme linked immunosorbent assay method. Compared with LC, apoptotic indices in HCC tissues were significantly reduced , expression of Fas was decreased , and that of FasL was increased . Serum sFas levels in HCC patients were significantly higher than those in normal controls. Down regulation of Fas expression, up regulation of FasL expression in hepatocytes and elevation of sFas level in serum might contribute to tumor escape from immune surveillance of the body. Apoptosis and the Fas system are significantly involved in the process of liver cirrhosis converting into hepatocellular carcinoma.展开更多
AIM： To determine the role of Fas/Fas ligand （FasL） in the immune escape of colon cancer cells. METHODS： Immunohistochemistry was used to observe the expression of Fas and FasL in the tissues of colon cancer patie...AIM： To determine the role of Fas/Fas ligand （FasL） in the immune escape of colon cancer cells. METHODS： Immunohistochemistry was used to observe the expression of Fas and FasL in the tissues of colon cancer patients. In situ hybridization was used to detect the localization of FasL mRNA expression in cancer tissues. Terminal deoxynucleotide transferase-mediated dUTP nick end labeling （TUNEL） assay and CD45 staining were performed to detect the apoptosis of tumor-infiltrating lymphocytes （TILs）. Co-culture assays of colon cancer cells （SW480） and Jurkat cells （Fas-sensitive cells） were performed to observe the counterattack of colon cancer cells to lymphocytes. RESULTS： Of 53 cases of colon carcinomas, 23 cases （43.4%） expressed Fas which was significantly lower as compared to the normal colonic mucosa （73.3%, P〈0.01）, and 45 cases （84.9%） of colon carcinomas expressed FasL, whereas only two cases （3.75%） in normal mucosa expressed FasL. FasL expression in the colon cancer cells was found to be associated with increased cell death of TIEs. The apoptotic rate of TIL in the FasL-positive staining regions of tumor cells was significantly higher than that in the FasL-negative staining region （54.84±2.79% vs 25.73±1.98%, P〈0.01）. The co-culture of SW480 cells and Jurkat cells confirmed the function of FasL on the SW480 cells. The apoptotic rates of Jurkat cells were found to be related with the amount of SW480 cells. CONCLUSION： Colon cancer cells can escape the immune surveillance and killing via decreasing Fas expression, and can counterattack the immune system via increasing FasL expression. Fas/FasL can serve as potential targets for effective antitumor therapy.展开更多
目的探讨炎调方调过Fas/Caspase-8信号通路减轻脓毒症急性胃肠损伤小鼠炎症的机制。方法取70只BALB/c小鼠随机分为空白组、假手术组和造模小鼠组。通过盲肠结扎穿孔术(cecum ligation and puncture,CLP)构建脓毒症急性胃肠损伤小鼠模型...目的探讨炎调方调过Fas/Caspase-8信号通路减轻脓毒症急性胃肠损伤小鼠炎症的机制。方法取70只BALB/c小鼠随机分为空白组、假手术组和造模小鼠组。通过盲肠结扎穿孔术(cecum ligation and puncture,CLP)构建脓毒症急性胃肠损伤小鼠模型,将造模成功的小鼠随机分为模型组,炎调方低、中、高剂量组,ROCK抑制剂组。苏木素-伊红(HE)染色观察小鼠回肠组织病理学改变;ELISA法检测各组小鼠血清IL-17、IL-23水平;蛋白印迹法检测回肠组织Fas/Caspase-8信号通路蛋白Fas、FADD和Caspase-8的相对表达;TUNEL染色法检测回肠组织细胞凋亡情况。结果与空白组相比,模型组小鼠回肠组织肠黏膜萎缩明显、绒毛排列杂乱,可见断裂、脱落,上皮细胞细胞坏死脱落,炎症细胞浸润明显,小鼠血清中IL-17、IL-23水平升高(P<0.05),回肠组织中Fas、FADD和Caspase-8蛋白的表达升高(P<0.05),肠上皮细胞呈现明显的凋亡现象(P<0.05)。与模型组相比,炎调方组小鼠的回肠组织病理学改变均得到不同程度的改善,血清中IL-17、IL-23水平降低(P<0.05),且回肠组织中Fas、FADD和Caspase-8蛋白的表达降低(P<0.05),肠上皮细胞凋亡减少(P<0.05)。结论炎调方可以减轻肠黏膜组织损伤和肠道组织炎症反应,可能是通过调控Fas/Caspase-8信号通路抑制脓毒症急性胃肠损伤小鼠的肠上皮细胞凋亡来发挥作用的。展开更多
AIM: To study the expression and serum level of HBxAg,Fas and FasL in tissues of HCC patients, and to assess the relationship between HBxAg and Fas/FasL system.METHODS: Tissues from 50 patients with HCC were tested fo...AIM: To study the expression and serum level of HBxAg,Fas and FasL in tissues of HCC patients, and to assess the relationship between HBxAg and Fas/FasL system.METHODS: Tissues from 50 patients with HCC were tested for the expression of HBxAg, Fas and FasL by S-P immunohistochemistry. Serum levels of sFas/sFasL and HBsAg/HBeAg were measured by ELISA assay. HBV X gene was detected by PCR in serum and confirmed by automatic sequencing. Fifty cases of liver cirrhosis and 30 normal controls were involved in serum analysis.RESULTS: The expression of HBxAg, Fas and FasL in carcinoma tissues was 96 %, 84 % and 98 %, respectively.Staining of HBxAg, Fas and FasL was observed predominately in cytoplasms, no significant difference was found in intensity between HBxAg, Fas and FasL (P＞0.05). HBxAg, Fas and FasL might express in the same area of carcinoma tissues and this co-expression could be found in most patients with HCC. The mean levels of sFas in serum from HCC, cirrhosis and normal controls were 762.29±391.56 μg@ L-1 835.36±407.33 μg@L-1 and 238.27±135.29 μg@L-1. The mean levels of sFasL in serum from HCC, cirrhosis and normal controls were 156.36±9.61iμg@ L-1, 173.63±18.74 μg@L-1 and 121.96±7.83 μg@ L-1.Statistical analysis showed that both sFas and sFasL in HCC and cirrhosis patients were significantly higher than those in normal controls (P＜0.01). Serum HBV X gene was found in 32 % of HCC patients and ,46 % of cirrhotic patients.There was no significant relationship between serum level of sFas/sFasL and serum X gene detection (P＞0.05). Eight percent of HCC patients with negative HBsAg and HBeAg in serum might have X gene in serum and HBxAg expression in carcinoma tissues.CONCLUSION: Our data suggest that HBxAg and Fas/FasL system plays an important role in the development of human HCC. Expression of HBxAg can leads to expression of Fas/FasL system which and reverse apoptosis of hepatocellular carcinoma induced by FasL.展开更多
目的·检测Fas相关死亡结构域蛋白(Fas-associated protein with death domain,FADD)在头颈部鳞状细胞癌(head and neck squamous cell carcinoma,HNSCC)中的表达水平,并探究FADD促进HNSCC细胞增殖的分子机制。方法·利用GEPIA ...目的·检测Fas相关死亡结构域蛋白(Fas-associated protein with death domain,FADD)在头颈部鳞状细胞癌(head and neck squamous cell carcinoma,HNSCC)中的表达水平,并探究FADD促进HNSCC细胞增殖的分子机制。方法·利用GEPIA 2数据库分析肿瘤组织中FADD表达水平及其与预后的关系;通过对HNSCC组织进行免疫组织化学染色(immunohistochemistry staining,IHC),探究FADD在正常、不典型增生和肿瘤组织中的表达水平变化;构建稳定低表达FADD的人HNSCC Fadu、HSC3细胞株,并通过蛋白印迹实验和实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)方法进行验证;使用LiveCyte活细胞追踪系统、克隆形成、细胞活力检测等方法探究FADD对HNSCC细胞增殖水平的调控作用;使用免疫共沉淀串联质谱(co-immunoprecipitation mass spectrum,Co-IP/MS)鉴定与FADD发生相互作用的蛋白,并应用CRISPR/Cas9技术、LiveCyte活细胞追踪系统、蛋白印迹实验等方法对与FADD相互作用的蛋白进行进一步机制研究。结果·数据库分析显示FADD在头颈鳞癌中显著高表达,并与患者不良预后相关。免疫组化染色表明FADD在HNSCC患者正常组织、不典型增生及肿瘤组织中的表达水平呈现递增趋势。在HNSCC细胞中敲低FADD后,与对照组相比,细胞的增殖能力显著降低,形成克隆数减少。Co-IP/MS结果显示,FADD与CUT样同源盒1(CUT-like homeobox 1,CUX1)蛋白存在相互作用,敲低FADD后CUX1表达水平升高。同时,在HNSCC细胞中敲除CUX1能够显著促进肿瘤细胞增殖能力。敲除CUX1可部分逆转FADD低表达引起的增殖抑制。结论·FADD在HNSCC中具有显著促癌作用,并与不良预后相关。FADD可通过与CUX1发生相互作用降低其表达水平进一步调控肿瘤细胞的增殖能力。展开更多
基金This project was supported by Shanghai Science and Tech-nology Development Foundation(No.98XD140 2 2 )
文摘To investigate the roles of apoptosis and the Fas system in the process of liver cirrhosis converting into hepatocellular carcinoma , expression of Fas and Fas ligand in 49 LC and 36 HCC samples was detected by immunohistochemical method. Apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling method. Serum soluble Fas levels in 28 cases of LC and 27 cases of HCC were measured by enzyme linked immunosorbent assay method. Compared with LC, apoptotic indices in HCC tissues were significantly reduced , expression of Fas was decreased , and that of FasL was increased . Serum sFas levels in HCC patients were significantly higher than those in normal controls. Down regulation of Fas expression, up regulation of FasL expression in hepatocytes and elevation of sFas level in serum might contribute to tumor escape from immune surveillance of the body. Apoptosis and the Fas system are significantly involved in the process of liver cirrhosis converting into hepatocellular carcinoma.
文摘AIM： To determine the role of Fas/Fas ligand （FasL） in the immune escape of colon cancer cells. METHODS： Immunohistochemistry was used to observe the expression of Fas and FasL in the tissues of colon cancer patients. In situ hybridization was used to detect the localization of FasL mRNA expression in cancer tissues. Terminal deoxynucleotide transferase-mediated dUTP nick end labeling （TUNEL） assay and CD45 staining were performed to detect the apoptosis of tumor-infiltrating lymphocytes （TILs）. Co-culture assays of colon cancer cells （SW480） and Jurkat cells （Fas-sensitive cells） were performed to observe the counterattack of colon cancer cells to lymphocytes. RESULTS： Of 53 cases of colon carcinomas, 23 cases （43.4%） expressed Fas which was significantly lower as compared to the normal colonic mucosa （73.3%, P〈0.01）, and 45 cases （84.9%） of colon carcinomas expressed FasL, whereas only two cases （3.75%） in normal mucosa expressed FasL. FasL expression in the colon cancer cells was found to be associated with increased cell death of TIEs. The apoptotic rate of TIL in the FasL-positive staining regions of tumor cells was significantly higher than that in the FasL-negative staining region （54.84±2.79% vs 25.73±1.98%, P〈0.01）. The co-culture of SW480 cells and Jurkat cells confirmed the function of FasL on the SW480 cells. The apoptotic rates of Jurkat cells were found to be related with the amount of SW480 cells. CONCLUSION： Colon cancer cells can escape the immune surveillance and killing via decreasing Fas expression, and can counterattack the immune system via increasing FasL expression. Fas/FasL can serve as potential targets for effective antitumor therapy.
文摘目的探讨炎调方调过Fas/Caspase-8信号通路减轻脓毒症急性胃肠损伤小鼠炎症的机制。方法取70只BALB/c小鼠随机分为空白组、假手术组和造模小鼠组。通过盲肠结扎穿孔术(cecum ligation and puncture,CLP)构建脓毒症急性胃肠损伤小鼠模型,将造模成功的小鼠随机分为模型组,炎调方低、中、高剂量组,ROCK抑制剂组。苏木素-伊红(HE)染色观察小鼠回肠组织病理学改变;ELISA法检测各组小鼠血清IL-17、IL-23水平;蛋白印迹法检测回肠组织Fas/Caspase-8信号通路蛋白Fas、FADD和Caspase-8的相对表达;TUNEL染色法检测回肠组织细胞凋亡情况。结果与空白组相比,模型组小鼠回肠组织肠黏膜萎缩明显、绒毛排列杂乱,可见断裂、脱落,上皮细胞细胞坏死脱落,炎症细胞浸润明显,小鼠血清中IL-17、IL-23水平升高(P<0.05),回肠组织中Fas、FADD和Caspase-8蛋白的表达升高(P<0.05),肠上皮细胞呈现明显的凋亡现象(P<0.05)。与模型组相比,炎调方组小鼠的回肠组织病理学改变均得到不同程度的改善,血清中IL-17、IL-23水平降低(P<0.05),且回肠组织中Fas、FADD和Caspase-8蛋白的表达降低(P<0.05),肠上皮细胞凋亡减少(P<0.05)。结论炎调方可以减轻肠黏膜组织损伤和肠道组织炎症反应,可能是通过调控Fas/Caspase-8信号通路抑制脓毒症急性胃肠损伤小鼠的肠上皮细胞凋亡来发挥作用的。
基金the Science Foundation of Fujian Province,No.99-Z-162
文摘AIM: To study the expression and serum level of HBxAg,Fas and FasL in tissues of HCC patients, and to assess the relationship between HBxAg and Fas/FasL system.METHODS: Tissues from 50 patients with HCC were tested for the expression of HBxAg, Fas and FasL by S-P immunohistochemistry. Serum levels of sFas/sFasL and HBsAg/HBeAg were measured by ELISA assay. HBV X gene was detected by PCR in serum and confirmed by automatic sequencing. Fifty cases of liver cirrhosis and 30 normal controls were involved in serum analysis.RESULTS: The expression of HBxAg, Fas and FasL in carcinoma tissues was 96 %, 84 % and 98 %, respectively.Staining of HBxAg, Fas and FasL was observed predominately in cytoplasms, no significant difference was found in intensity between HBxAg, Fas and FasL (P＞0.05). HBxAg, Fas and FasL might express in the same area of carcinoma tissues and this co-expression could be found in most patients with HCC. The mean levels of sFas in serum from HCC, cirrhosis and normal controls were 762.29±391.56 μg@ L-1 835.36±407.33 μg@L-1 and 238.27±135.29 μg@L-1. The mean levels of sFasL in serum from HCC, cirrhosis and normal controls were 156.36±9.61iμg@ L-1, 173.63±18.74 μg@L-1 and 121.96±7.83 μg@ L-1.Statistical analysis showed that both sFas and sFasL in HCC and cirrhosis patients were significantly higher than those in normal controls (P＜0.01). Serum HBV X gene was found in 32 % of HCC patients and ,46 % of cirrhotic patients.There was no significant relationship between serum level of sFas/sFasL and serum X gene detection (P＞0.05). Eight percent of HCC patients with negative HBsAg and HBeAg in serum might have X gene in serum and HBxAg expression in carcinoma tissues.CONCLUSION: Our data suggest that HBxAg and Fas/FasL system plays an important role in the development of human HCC. Expression of HBxAg can leads to expression of Fas/FasL system which and reverse apoptosis of hepatocellular carcinoma induced by FasL.
文摘目的·检测Fas相关死亡结构域蛋白(Fas-associated protein with death domain,FADD)在头颈部鳞状细胞癌(head and neck squamous cell carcinoma,HNSCC)中的表达水平,并探究FADD促进HNSCC细胞增殖的分子机制。方法·利用GEPIA 2数据库分析肿瘤组织中FADD表达水平及其与预后的关系;通过对HNSCC组织进行免疫组织化学染色(immunohistochemistry staining,IHC),探究FADD在正常、不典型增生和肿瘤组织中的表达水平变化;构建稳定低表达FADD的人HNSCC Fadu、HSC3细胞株,并通过蛋白印迹实验和实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)方法进行验证;使用LiveCyte活细胞追踪系统、克隆形成、细胞活力检测等方法探究FADD对HNSCC细胞增殖水平的调控作用;使用免疫共沉淀串联质谱(co-immunoprecipitation mass spectrum,Co-IP/MS)鉴定与FADD发生相互作用的蛋白,并应用CRISPR/Cas9技术、LiveCyte活细胞追踪系统、蛋白印迹实验等方法对与FADD相互作用的蛋白进行进一步机制研究。结果·数据库分析显示FADD在头颈鳞癌中显著高表达,并与患者不良预后相关。免疫组化染色表明FADD在HNSCC患者正常组织、不典型增生及肿瘤组织中的表达水平呈现递增趋势。在HNSCC细胞中敲低FADD后,与对照组相比,细胞的增殖能力显著降低,形成克隆数减少。Co-IP/MS结果显示,FADD与CUT样同源盒1(CUT-like homeobox 1,CUX1)蛋白存在相互作用,敲低FADD后CUX1表达水平升高。同时,在HNSCC细胞中敲除CUX1能够显著促进肿瘤细胞增殖能力。敲除CUX1可部分逆转FADD低表达引起的增殖抑制。结论·FADD在HNSCC中具有显著促癌作用,并与不良预后相关。FADD可通过与CUX1发生相互作用降低其表达水平进一步调控肿瘤细胞的增殖能力。
