AIM： To explore the effect of silencing of signal transducer and activator of transcription 3 （STAT3） expression by RNA interference （RNAi） on growth of human hepatocellular carcinoma （HCC） in tumorbearing nude...AIM： To explore the effect of silencing of signal transducer and activator of transcription 3 （STAT3） expression by RNA interference （RNAi） on growth of human hepatocellular carcinoma （HCC） in tumorbearing nude mice in vivo.METHODS： To construct the recombinant plasmid of pSilencer 3.0-H1-STAT3-siRNA-GFP （pSHI-siRNA- STAT3） and establish the tumor-bearing nude mouse model of the HCC cell line SMMC7721, we used intratumoral injection together with electroblotting to transfect the recombinant plasmid pSHI-siRNA- STAT3 into the transplanted tumor. The weight of the nude mice and tumor volumes were recorded. STAT3 gene transcription was detected by semi-quantitative reverse transcription polymerase chain reaction （RT- PCR）. Level of protein expression and location of STAT3 were determined by Western blotting and immunohistochemical staining. STAT3-related genes such as survivin, c-myc, VEGF, p53 and caspase3 mRNA and protein expression were detected in tumor tissues at the same time. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay was used to detect apoptosis of tumor cells.RESULTS： The weight of the treated nude mice increased, and the tumor volume decreased markedly compared with those of the mock-treated and negative control groups （P 〈 0.01）. The results of RT-PCR and Western blotting showed that mRNA and protein levels of STAT3 declined markedly in the treated group. The change in STAT3-related gene expression in tumor tissues at the mRNA and protein level also varied, the expression of survivin, VEGF and c-myc were obviously reduced, and expression of p53 and caspase3 increased （P 〈 0.01）. Most of the tumor tissue ceils in the treated group developed apoptosis that was detected by TUNEL assay.CONCLUSION： Silencing of STAT3 expression by RNAi significantly inhibits expression of STAT3 mRNA and protein, and suppresses growth of human HCC in tumor-bearing nude mice. The mechanism may be related to down-regulation of survivin, VEGF and c-myc and up-regulation of p53 and caspase3 expression. Accordingly, the STAT3 gene may act as an important and effective target in gene therapy of HCC.展开更多
[目的]研究鹿红方改善心肌梗死后心肌纤维化的作用机制。[方法]采用冠状动脉结扎法制备心肌梗死后心肌纤维化模型。将60只SD大鼠随机分配至假手术组、模型组、鹿红方组以及培哚普利组,干预4周后用心脏彩色多普勒超声检查测量心脏结构和...[目的]研究鹿红方改善心肌梗死后心肌纤维化的作用机制。[方法]采用冠状动脉结扎法制备心肌梗死后心肌纤维化模型。将60只SD大鼠随机分配至假手术组、模型组、鹿红方组以及培哚普利组,干预4周后用心脏彩色多普勒超声检查测量心脏结构和左室射血分数,苏木精-伊红(hematoxylin-eosin,HE)染色、Masson和天狼星红染色观察心脏组织纤维化病理改变,免疫印迹检测心脏信号转导及转录激活因子3(signal transducer and activator of transcription 3,STAT3)分泌情况,酶联免疫吸附分析(enzymelinked immunosorbent assay,ELISA)检测血清促纤维化因子结缔组织生长因子(connective tissue growth factor,CTGF)、血小板反应蛋白-1(thrombospondin-1,TSP-1)、基质金属蛋白酶抑制剂-1(tissue inhibitor of metalloproteinase-1,TIMP-1)的水平。[结果]心脏彩色多普勒超声检查提示,与模型组比较,鹿红方和培哚普利干预均可抑制左室舒张末容积和左室舒张末内径的增大,而且使左室射血分数明显提升(P<0.05)。与培哚普利组比较,鹿红方组改善心室不良扩大与提高左室射血分数作用相近。HE染色结果显示,与模型组比较,鹿红方组和培哚普利组异常形态的心肌细胞数目明显减少,细胞间质肿胀减轻,炎症细胞浸润减少。Masson和天狼星红染色结果显示,与模型组比较,鹿红方组和培哚普利组干预均可减轻心肌纤维化病变程度。鹿红方组改善上述心肌病理改变和减轻心肌纤维化作用与培哚普利组相近。免疫印迹检测显示,与模型组比较,鹿红方和培哚普利干预均可增加心肌STAT3分泌(P<0.05),鹿红方组促进心肌分泌STAT3的作用与培哚普利组相近,差异无统计学意义(P>0.05)。ELISA结果显示,与模型组比较,鹿红方组与培哚普利组CTGF、TSP-1、TIMP-1水平显著降低(P<0.05),鹿红方组抑制CTGF、TSP-1和TIMP-1分泌作用与培哚普利组相近,差异无统计学意义(P>0.05)。[结论]鹿红方可显著抑制心肌梗死后心肌纤维化,其机制与上调STAT3水平,抑制促纤维化因子CTGF、TSP-1和TIMP-1分泌有关。展开更多
基金Supported by The Science and Technology Fund of Jilin Province,No. 200505219
文摘AIM： To explore the effect of silencing of signal transducer and activator of transcription 3 （STAT3） expression by RNA interference （RNAi） on growth of human hepatocellular carcinoma （HCC） in tumorbearing nude mice in vivo.METHODS： To construct the recombinant plasmid of pSilencer 3.0-H1-STAT3-siRNA-GFP （pSHI-siRNA- STAT3） and establish the tumor-bearing nude mouse model of the HCC cell line SMMC7721, we used intratumoral injection together with electroblotting to transfect the recombinant plasmid pSHI-siRNA- STAT3 into the transplanted tumor. The weight of the nude mice and tumor volumes were recorded. STAT3 gene transcription was detected by semi-quantitative reverse transcription polymerase chain reaction （RT- PCR）. Level of protein expression and location of STAT3 were determined by Western blotting and immunohistochemical staining. STAT3-related genes such as survivin, c-myc, VEGF, p53 and caspase3 mRNA and protein expression were detected in tumor tissues at the same time. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay was used to detect apoptosis of tumor cells.RESULTS： The weight of the treated nude mice increased, and the tumor volume decreased markedly compared with those of the mock-treated and negative control groups （P 〈 0.01）. The results of RT-PCR and Western blotting showed that mRNA and protein levels of STAT3 declined markedly in the treated group. The change in STAT3-related gene expression in tumor tissues at the mRNA and protein level also varied, the expression of survivin, VEGF and c-myc were obviously reduced, and expression of p53 and caspase3 increased （P 〈 0.01）. Most of the tumor tissue ceils in the treated group developed apoptosis that was detected by TUNEL assay.CONCLUSION： Silencing of STAT3 expression by RNAi significantly inhibits expression of STAT3 mRNA and protein, and suppresses growth of human HCC in tumor-bearing nude mice. The mechanism may be related to down-regulation of survivin, VEGF and c-myc and up-regulation of p53 and caspase3 expression. Accordingly, the STAT3 gene may act as an important and effective target in gene therapy of HCC.
文摘[目的]研究鹿红方改善心肌梗死后心肌纤维化的作用机制。[方法]采用冠状动脉结扎法制备心肌梗死后心肌纤维化模型。将60只SD大鼠随机分配至假手术组、模型组、鹿红方组以及培哚普利组,干预4周后用心脏彩色多普勒超声检查测量心脏结构和左室射血分数,苏木精-伊红(hematoxylin-eosin,HE)染色、Masson和天狼星红染色观察心脏组织纤维化病理改变,免疫印迹检测心脏信号转导及转录激活因子3(signal transducer and activator of transcription 3,STAT3)分泌情况,酶联免疫吸附分析(enzymelinked immunosorbent assay,ELISA)检测血清促纤维化因子结缔组织生长因子(connective tissue growth factor,CTGF)、血小板反应蛋白-1(thrombospondin-1,TSP-1)、基质金属蛋白酶抑制剂-1(tissue inhibitor of metalloproteinase-1,TIMP-1)的水平。[结果]心脏彩色多普勒超声检查提示,与模型组比较,鹿红方和培哚普利干预均可抑制左室舒张末容积和左室舒张末内径的增大,而且使左室射血分数明显提升(P<0.05)。与培哚普利组比较,鹿红方组改善心室不良扩大与提高左室射血分数作用相近。HE染色结果显示,与模型组比较,鹿红方组和培哚普利组异常形态的心肌细胞数目明显减少,细胞间质肿胀减轻,炎症细胞浸润减少。Masson和天狼星红染色结果显示,与模型组比较,鹿红方组和培哚普利组干预均可减轻心肌纤维化病变程度。鹿红方组改善上述心肌病理改变和减轻心肌纤维化作用与培哚普利组相近。免疫印迹检测显示,与模型组比较,鹿红方和培哚普利干预均可增加心肌STAT3分泌(P<0.05),鹿红方组促进心肌分泌STAT3的作用与培哚普利组相近,差异无统计学意义(P>0.05)。ELISA结果显示,与模型组比较,鹿红方组与培哚普利组CTGF、TSP-1、TIMP-1水平显著降低(P<0.05),鹿红方组抑制CTGF、TSP-1和TIMP-1分泌作用与培哚普利组相近,差异无统计学意义(P>0.05)。[结论]鹿红方可显著抑制心肌梗死后心肌纤维化,其机制与上调STAT3水平,抑制促纤维化因子CTGF、TSP-1和TIMP-1分泌有关。
