目的:探讨bi kuni n对前列腺癌细胞侵袭能力的影响及其相关的分子机制。方法:利用bi kuni n处理前列腺癌细胞系PC3,采用t r ans wel l实验检测其对细胞侵袭能力的影响。同时,利用west ern印迹技术检测PI3K/AKT信号通路在此过程中的作用...目的:探讨bi kuni n对前列腺癌细胞侵袭能力的影响及其相关的分子机制。方法:利用bi kuni n处理前列腺癌细胞系PC3,采用t r ans wel l实验检测其对细胞侵袭能力的影响。同时,利用west ern印迹技术检测PI3K/AKT信号通路在此过程中的作用。结果:Bi kuni n能够显著降低PC3细胞的侵袭能力;Bi kuni n能够抑制PI3K和AKT的活性;PI3K和AKT的抑制剂能够抑制PC3细胞的侵袭;组成型活化的PI3K能够阻断bi kuni n的抑制作用。结论:Bi kuni n可能通过调节PI3K/AKT信号通路来抑制前列腺癌细胞的侵袭;Bi kuni n具有一定的临床应用前景。展开更多
The light chain of inter-α inhibitor, also known as bikunin or urinary trypsin inhibitor, is composed of two tandemly arranged Kunitz-type protease inhibitor domains. The second domain of bikunin has factor Xa inhibi...The light chain of inter-α inhibitor, also known as bikunin or urinary trypsin inhibitor, is composed of two tandemly arranged Kunitz-type protease inhibitor domains. The second domain of bikunin has factor Xa inhibitory activity which previously was enhanced by mutating two amino acids, glutamine 19 and tyrosine 46 to lysine and aspartate, respectively. In this study, we tried to potentiate its inhibitory activity against leukocyte elastase. A molecular docking model of the second domain of bikunin with leukocyte elastase revealed that P5 arginine 11 was a candidate residue for a third substitution. We generated six triple point mutants using site-directed mutagenesis, compared their leukocyte elastase-inhibitory activities, and selected the most potent variant with arginine 11 substituted to serine. The IC50 values for factor XIa, factor Xa, and leukocyte elastase were 182, 302, and 273 nM, respectively. Moreover, this triple point mutant prolonged the activated partial thromboplastin time and moderately reduced leukocyte elastase-induced endothelial injury. Additionally, favorable conformations created by these mutations were speculated using the structure of the Kunitz protease inhibitor domain of protease nexin 2 complexed with factor XIa as a reference. We discovered a novel triple point mutant of the second domain of bikunin that has potent inhibitory activities against factor XIa, factor Xa, and leukocyte elastase. This variant exhibited anticoagulant activity in plasma and suppressed endothelial cell injury.展开更多
目的从蛋白水平探讨Bikunin,又称尿胰蛋白酶抑制剂(urinary trypsin inhibitor,UTI)在上皮性卵巢癌浸润、转移中的作用,及其在组织中的分布情况、与预后的关系。方法应用免疫组化法检测80份上皮性卵巢癌、20份良性卵巢肿瘤标本中Bikuni...目的从蛋白水平探讨Bikunin,又称尿胰蛋白酶抑制剂(urinary trypsin inhibitor,UTI)在上皮性卵巢癌浸润、转移中的作用,及其在组织中的分布情况、与预后的关系。方法应用免疫组化法检测80份上皮性卵巢癌、20份良性卵巢肿瘤标本中Bikunin蛋白表达,并结合临床病理因素、预后进行分析。结果上皮性卵巢癌、良性卵巢肿瘤标本中Bikunin阳性率分别为55.0%、20.0%,两者差异有统计学意义(χ2=7.853,P=0.005)。上皮性卵巢癌中,Bikunin阳性与国际妇产科联盟(International Federation of Gynecology and Obstetrics,FIGO)分期早有显著性相关,差异有统计学意义(χ2=5.241,P=0.022),与上述其他临床病理因素差异无统计学意义(P>0.05)。多因素Cox回归模型显示,Bikunin表达是总生存的独立危险因素。结论上皮性卵巢癌中Bikunin表达上调,Bikunin有可能作为预测上皮性卵巢癌预后的参考指标。展开更多
文摘目的:探讨bi kuni n对前列腺癌细胞侵袭能力的影响及其相关的分子机制。方法:利用bi kuni n处理前列腺癌细胞系PC3,采用t r ans wel l实验检测其对细胞侵袭能力的影响。同时,利用west ern印迹技术检测PI3K/AKT信号通路在此过程中的作用。结果:Bi kuni n能够显著降低PC3细胞的侵袭能力;Bi kuni n能够抑制PI3K和AKT的活性;PI3K和AKT的抑制剂能够抑制PC3细胞的侵袭;组成型活化的PI3K能够阻断bi kuni n的抑制作用。结论:Bi kuni n可能通过调节PI3K/AKT信号通路来抑制前列腺癌细胞的侵袭;Bi kuni n具有一定的临床应用前景。
文摘The light chain of inter-α inhibitor, also known as bikunin or urinary trypsin inhibitor, is composed of two tandemly arranged Kunitz-type protease inhibitor domains. The second domain of bikunin has factor Xa inhibitory activity which previously was enhanced by mutating two amino acids, glutamine 19 and tyrosine 46 to lysine and aspartate, respectively. In this study, we tried to potentiate its inhibitory activity against leukocyte elastase. A molecular docking model of the second domain of bikunin with leukocyte elastase revealed that P5 arginine 11 was a candidate residue for a third substitution. We generated six triple point mutants using site-directed mutagenesis, compared their leukocyte elastase-inhibitory activities, and selected the most potent variant with arginine 11 substituted to serine. The IC50 values for factor XIa, factor Xa, and leukocyte elastase were 182, 302, and 273 nM, respectively. Moreover, this triple point mutant prolonged the activated partial thromboplastin time and moderately reduced leukocyte elastase-induced endothelial injury. Additionally, favorable conformations created by these mutations were speculated using the structure of the Kunitz protease inhibitor domain of protease nexin 2 complexed with factor XIa as a reference. We discovered a novel triple point mutant of the second domain of bikunin that has potent inhibitory activities against factor XIa, factor Xa, and leukocyte elastase. This variant exhibited anticoagulant activity in plasma and suppressed endothelial cell injury.
文摘目的从蛋白水平探讨Bikunin,又称尿胰蛋白酶抑制剂(urinary trypsin inhibitor,UTI)在上皮性卵巢癌浸润、转移中的作用,及其在组织中的分布情况、与预后的关系。方法应用免疫组化法检测80份上皮性卵巢癌、20份良性卵巢肿瘤标本中Bikunin蛋白表达,并结合临床病理因素、预后进行分析。结果上皮性卵巢癌、良性卵巢肿瘤标本中Bikunin阳性率分别为55.0%、20.0%,两者差异有统计学意义(χ2=7.853,P=0.005)。上皮性卵巢癌中,Bikunin阳性与国际妇产科联盟(International Federation of Gynecology and Obstetrics,FIGO)分期早有显著性相关,差异有统计学意义(χ2=5.241,P=0.022),与上述其他临床病理因素差异无统计学意义(P>0.05)。多因素Cox回归模型显示,Bikunin表达是总生存的独立危险因素。结论上皮性卵巢癌中Bikunin表达上调,Bikunin有可能作为预测上皮性卵巢癌预后的参考指标。
