BACKGROUND: Notch signaling is critical to physiologic angiogenesis and has been implicated in tumor angiogenesis and metastasis. Notch signaling was reported to exert either oncogenic or tumor-suppressive function in...BACKGROUND: Notch signaling is critical to physiologic angiogenesis and has been implicated in tumor angiogenesis and metastasis. Notch signaling was reported to exert either oncogenic or tumor-suppressive function in hepatocellular carcinoma (HCC) tumorigenesis. However, the prognostic significance of Notch receptors in HCC remains uncertain. In this study, we investigated the roles of Notch receptors in the prognosis of HCC. METHODS: We investigated the expressions of Notch receptors in tumor tissue microarrays of 288 patients with primary HCC who had undergone curative resection using immunohistochemistry. Additionally, prognostic factors of HCC were examined by univariate and multivariate analyses. The median follow-up period was 97.1 months. Tumor recurrence was detected in 189 patients (65.6%), and 99 (34.4%) died of HCC. RESULTS: Cytoplasmic expression of Notch1, cytoplasmic expression of Notch3, coexistent nuclear expression of Notch3, and cytoplasmic Notch4 overexpression were observed in 145 (50.3%), 60 (20.8%), 17 (5.9%), and 172 (59.7%) of the 288 HCCs, respectively. Multivariate analyses revealed that Notch1 expression (P=0.029), Edmondson grade Ⅲ (P=0.038), and higher BCLC stage (P【0.001) were independent predictors of shorter disease-free survival. Cytoplasmic Notch3 expression tended to be an independent predictor of shorter disease-free survival (P=0.055). Notch1 expression (P=0.039), Notch4 overexpression (P=0.012), and higher BCLC stage (P【0.001) were independent predictors of shorter disease-specific survival. On univariate analysis, Notch1 expression tended to show an unfavorable influence on disease-specific survival (P=0.063) and Notch4 overexpression did not show an unfavorable influence on disease-specific survival (P=0.103). CONCLUSIONS: Notch1 expression might be an independent predictor of both shorter disease-free survival and shorter disease-specific survival in HCC patients after curative resection. Notch4 overexpression might be an independent predictor of shorter disease-s展开更多
Vasculogenic mimicry（VM） is a process by which aggressive tumor cells generate non-endothelial cell-lined channels in malignant tumors including hepatocellular carcinoma（HCC）. It has provided new insights into tum...Vasculogenic mimicry（VM） is a process by which aggressive tumor cells generate non-endothelial cell-lined channels in malignant tumors including hepatocellular carcinoma（HCC）. It has provided new insights into tumor behavior and has surfaced as a potential target for drug therapy. The molecular events underlying the process of VM formation are still poorly understood. In this study, we attempted to elucidate the relationship between Notch4 and VM formation in HCC. An effective si RNA lentiviral vector targeting Notch4 was constructed and transfected into Bel7402, a HCC cell line. VM networks were observed with a microscope in a 3 dimensional cell culture system. Cell migration and invasion were evaluated using wound healing and transwell assays. Matrix metalloproteinases（MMPs） activity was detected by gelatin zymography. Furthermore, the role of Notch4 inhibition in Bel7402 cells in vivo was examined in subcutaneous xenograft tumor model of mice. The results showed that downregulation of Notch4 destroyed VM network formation and inhibited migration and invasion of tumor cells in vitro（P〈0.05）. In vivo, tumor growth was also inhibited in subcutaneous xenograft model（P〈0.05）. The potential mechanisms might be related with down-regulation of MT1-MMP, MMP-2, MMP-9 expression and inhibition of the activation of MMP2 and MMP9. These results indicated that Notch4 may play an important role in VM formation and tumor invasion in HCC. Related molecular pathways may be used as novel therapeutic targets for HCC antiangiogenesis therapy.展开更多
OBJECTIVE： To investigate, in terms of Notch signaling pathway, the effect on pancreatic cancer of the extract of an anti-tumor prescription -- Qingyihuaji formula （QYHJ） -- from Traditional Chinese Medicine （TCM...OBJECTIVE： To investigate, in terms of Notch signaling pathway, the effect on pancreatic cancer of the extract of an anti-tumor prescription -- Qingyihuaji formula （QYHJ） -- from Traditional Chinese Medicine （TCM）.METHODS： Nude mice were implanted subcutaneously with human pancreatic cancer cell line SW1990 and then randomly divided into four groups： Control, QYHJ extract, Gemcitabine, and Combination of QYHJ extract and gemcitabine. Treatments were given for 21 days and tumor growth was evaluated simultaneously. Then, expression of Notch receptors （Notch-I, Notch-2, Notch-3, and Notch-4） and their Jagged ligands （Jagged-1 and Jagged-2） in dissected tumor tissue were detected by real-time quantitative reverse transcription-polymerase chain reaction and Western blot. Finally, immunohistochemistry was performed to detect CD133, a marker of pancreatic cancer stem cells （CSCs）, to evaluate the impact of QYHJ extract on pancreatic CSCs.RESULTS： QYHJ extract treatment effectively inhib- ited the tumor growth in nude mice. The expression of both Notch-4 and Jagged-1 were decreased significantly in QYHJ treatment groups （P 〈 0.05）, while gemcitabine alone had no significant effect in down-regulating Jagged-1 （P 〉 0.05）. No significant difference was observed in the ex- pression of Notch-1, Notch-2, Notch-3, and Jagged-2 between three treatment groups and control group （P 〉 0.05）. Moreover, immunohistochemical analysis showed that the number of CD133 positive cells was significantly reduced by QYHJ treatment （P 〈 0.05）, and the combined treatment was more effective than gemcitabine alone （P 〈 0.05）.CONCLUSION： The role of the extract in pancreatic cancer treatment was associated with down-regulation of Notch-4 and Jagged-1 in Notch signaling pathway. The extract could enhance the antitumor activity of gemcitabine and was more effective than gemcitabine in regulating Notch signaling pathway to some extent.展开更多
目的研究Notch4受体激活后对慢性髓系白血病细胞株K562增殖的影响及可能的作用机制。方法用脂质体分别将携带Notch4胞内段(ICN4)的质粒pc DNA 3.1-ICN4及空载体质粒pc DNA 3.1转染入K562细胞,用G418筛选出稳定表达ICN4的细胞株。观察K56...目的研究Notch4受体激活后对慢性髓系白血病细胞株K562增殖的影响及可能的作用机制。方法用脂质体分别将携带Notch4胞内段(ICN4)的质粒pc DNA 3.1-ICN4及空载体质粒pc DNA 3.1转染入K562细胞,用G418筛选出稳定表达ICN4的细胞株。观察K562细胞ICN4转染后的形态变化,MTT法分析细胞增殖水平,RT-PCR法和Western blot法检测Notch4受体、Hes1、Hey1下游靶基因mRNA及蛋白的表达,流式细胞仪检测细胞周期分布。结果筛选出稳定表达ICN4的细胞株K562/ICN4细胞,与对照组及空载体组比较,ICN4转染组K562细胞数量减少,胞膜透亮度减小,核染色质固缩,核浆比例减低;ICN4转染后K562细胞生长受抑(P<0.05),且随时间延长而明显;Notch4受体及Hes1下游靶基因mRNA及蛋白表达水平相似,均较对照组及空载体组表达增强,但Hey1下游靶基因mRNA及蛋白表达水平未见明显变化;细胞周期检测结果示,转染48 h后细胞阻滞于G1期(P<0.05),S期细胞减少(P<0.05)。结论 Notch4受体激活可抑制K562细胞增殖,其机制可能是通过激活下游靶基因Hes1表达,而调控细胞于G1期而实现的。展开更多
文摘BACKGROUND: Notch signaling is critical to physiologic angiogenesis and has been implicated in tumor angiogenesis and metastasis. Notch signaling was reported to exert either oncogenic or tumor-suppressive function in hepatocellular carcinoma (HCC) tumorigenesis. However, the prognostic significance of Notch receptors in HCC remains uncertain. In this study, we investigated the roles of Notch receptors in the prognosis of HCC. METHODS: We investigated the expressions of Notch receptors in tumor tissue microarrays of 288 patients with primary HCC who had undergone curative resection using immunohistochemistry. Additionally, prognostic factors of HCC were examined by univariate and multivariate analyses. The median follow-up period was 97.1 months. Tumor recurrence was detected in 189 patients (65.6%), and 99 (34.4%) died of HCC. RESULTS: Cytoplasmic expression of Notch1, cytoplasmic expression of Notch3, coexistent nuclear expression of Notch3, and cytoplasmic Notch4 overexpression were observed in 145 (50.3%), 60 (20.8%), 17 (5.9%), and 172 (59.7%) of the 288 HCCs, respectively. Multivariate analyses revealed that Notch1 expression (P=0.029), Edmondson grade Ⅲ (P=0.038), and higher BCLC stage (P【0.001) were independent predictors of shorter disease-free survival. Cytoplasmic Notch3 expression tended to be an independent predictor of shorter disease-free survival (P=0.055). Notch1 expression (P=0.039), Notch4 overexpression (P=0.012), and higher BCLC stage (P【0.001) were independent predictors of shorter disease-specific survival. On univariate analysis, Notch1 expression tended to show an unfavorable influence on disease-specific survival (P=0.063) and Notch4 overexpression did not show an unfavorable influence on disease-specific survival (P=0.103). CONCLUSIONS: Notch1 expression might be an independent predictor of both shorter disease-free survival and shorter disease-specific survival in HCC patients after curative resection. Notch4 overexpression might be an independent predictor of shorter disease-s
基金supported by grants from the Sci-Tech Research Foundation of Fujian Province(No.2011J05067)the National Clinical Key Specialty Construction Project(General Surgery)of China(No.[2012]649)
文摘Vasculogenic mimicry（VM） is a process by which aggressive tumor cells generate non-endothelial cell-lined channels in malignant tumors including hepatocellular carcinoma（HCC）. It has provided new insights into tumor behavior and has surfaced as a potential target for drug therapy. The molecular events underlying the process of VM formation are still poorly understood. In this study, we attempted to elucidate the relationship between Notch4 and VM formation in HCC. An effective si RNA lentiviral vector targeting Notch4 was constructed and transfected into Bel7402, a HCC cell line. VM networks were observed with a microscope in a 3 dimensional cell culture system. Cell migration and invasion were evaluated using wound healing and transwell assays. Matrix metalloproteinases（MMPs） activity was detected by gelatin zymography. Furthermore, the role of Notch4 inhibition in Bel7402 cells in vivo was examined in subcutaneous xenograft tumor model of mice. The results showed that downregulation of Notch4 destroyed VM network formation and inhibited migration and invasion of tumor cells in vitro（P〈0.05）. In vivo, tumor growth was also inhibited in subcutaneous xenograft model（P〈0.05）. The potential mechanisms might be related with down-regulation of MT1-MMP, MMP-2, MMP-9 expression and inhibition of the activation of MMP2 and MMP9. These results indicated that Notch4 may play an important role in VM formation and tumor invasion in HCC. Related molecular pathways may be used as novel therapeutic targets for HCC antiangiogenesis therapy.
文摘目的探讨电针结合康复训练治疗对脑卒中大鼠神经功能及大鼠大脑皮层Notch1与Notch4表达的影响。方法 60只SD大鼠随机分为3组:假手术组(S)、模型组(M)与电针+康复训练组(E),采用大脑中动脉阻塞法(MCAO)建立局灶性脑缺血动物模型。采用Longa法评定实验大鼠的神经功能缺损程度,免疫组织化学方法与Western blot和real time PCR方法检测大鼠大脑皮层Notch1与Notch4表达。结果与模型组相比,电针+康复训练组大鼠在实施治疗后,Longa评分明显改善(P<0.01)。与假手术组相比,模型组大鼠大脑皮层Notch1与Notch4蛋白与mRNA表达水平显著升高(P<0.01);而与模型组相比,电针+康复训练组大鼠大脑皮层Notch1与Notch4蛋白与mRNA表达水平升高更为显著(P<0.01)。结论电针结合康复训练改善脑卒中大鼠的神经功能可与上调大鼠大脑皮层Notch1与Notch4的表达相关。
基金Supported by the National Natural Science Foundation of China(No.81173461)China Scholarship Council(No.201306100055)
文摘OBJECTIVE： To investigate, in terms of Notch signaling pathway, the effect on pancreatic cancer of the extract of an anti-tumor prescription -- Qingyihuaji formula （QYHJ） -- from Traditional Chinese Medicine （TCM）.METHODS： Nude mice were implanted subcutaneously with human pancreatic cancer cell line SW1990 and then randomly divided into four groups： Control, QYHJ extract, Gemcitabine, and Combination of QYHJ extract and gemcitabine. Treatments were given for 21 days and tumor growth was evaluated simultaneously. Then, expression of Notch receptors （Notch-I, Notch-2, Notch-3, and Notch-4） and their Jagged ligands （Jagged-1 and Jagged-2） in dissected tumor tissue were detected by real-time quantitative reverse transcription-polymerase chain reaction and Western blot. Finally, immunohistochemistry was performed to detect CD133, a marker of pancreatic cancer stem cells （CSCs）, to evaluate the impact of QYHJ extract on pancreatic CSCs.RESULTS： QYHJ extract treatment effectively inhib- ited the tumor growth in nude mice. The expression of both Notch-4 and Jagged-1 were decreased significantly in QYHJ treatment groups （P 〈 0.05）, while gemcitabine alone had no significant effect in down-regulating Jagged-1 （P 〉 0.05）. No significant difference was observed in the ex- pression of Notch-1, Notch-2, Notch-3, and Jagged-2 between three treatment groups and control group （P 〉 0.05）. Moreover, immunohistochemical analysis showed that the number of CD133 positive cells was significantly reduced by QYHJ treatment （P 〈 0.05）, and the combined treatment was more effective than gemcitabine alone （P 〈 0.05）.CONCLUSION： The role of the extract in pancreatic cancer treatment was associated with down-regulation of Notch-4 and Jagged-1 in Notch signaling pathway. The extract could enhance the antitumor activity of gemcitabine and was more effective than gemcitabine in regulating Notch signaling pathway to some extent.
文摘目的研究Notch4受体激活后对慢性髓系白血病细胞株K562增殖的影响及可能的作用机制。方法用脂质体分别将携带Notch4胞内段(ICN4)的质粒pc DNA 3.1-ICN4及空载体质粒pc DNA 3.1转染入K562细胞,用G418筛选出稳定表达ICN4的细胞株。观察K562细胞ICN4转染后的形态变化,MTT法分析细胞增殖水平,RT-PCR法和Western blot法检测Notch4受体、Hes1、Hey1下游靶基因mRNA及蛋白的表达,流式细胞仪检测细胞周期分布。结果筛选出稳定表达ICN4的细胞株K562/ICN4细胞,与对照组及空载体组比较,ICN4转染组K562细胞数量减少,胞膜透亮度减小,核染色质固缩,核浆比例减低;ICN4转染后K562细胞生长受抑(P<0.05),且随时间延长而明显;Notch4受体及Hes1下游靶基因mRNA及蛋白表达水平相似,均较对照组及空载体组表达增强,但Hey1下游靶基因mRNA及蛋白表达水平未见明显变化;细胞周期检测结果示,转染48 h后细胞阻滞于G1期(P<0.05),S期细胞减少(P<0.05)。结论 Notch4受体激活可抑制K562细胞增殖,其机制可能是通过激活下游靶基因Hes1表达,而调控细胞于G1期而实现的。
