目的探讨HAI-1表达水平与子宫内膜癌细胞株侵袭迁移能力的相关性。方法体外培养人子宫内膜癌细胞株,采用Real-time q PCR技术检测HEC-1A、HEC-1B和RL952三株子宫子宫内膜癌细胞中HAI-1 m RNA表达水平,应用细胞划痕实验及穿膜小室实验,...目的探讨HAI-1表达水平与子宫内膜癌细胞株侵袭迁移能力的相关性。方法体外培养人子宫内膜癌细胞株,采用Real-time q PCR技术检测HEC-1A、HEC-1B和RL952三株子宫子宫内膜癌细胞中HAI-1 m RNA表达水平,应用细胞划痕实验及穿膜小室实验,观察三株子宫内膜癌细胞体外的侵袭及迁移能力。结果 (1)HAI-1 m RNA在三株细胞中的相对表达量分别为(0.3042±0.0101)、(0.0032±0.0001)和(0.2580±0.0096),HAI-1蛋白在HEC-1A、HEC-1B和RL952中均呈阳性表达,但在HEC-1B中表达低于HEC-1A及RL952。(2)细胞划痕实验结果显示,HEC-1A、HEC-1B和RL952三组迁移距离分别为(173.30±7.33)μm、(437.00±8.72)μm和(135.30±6.89)μm。(3)侵袭实验结果显示,HEC-1A、HEC-1B和RL952三组穿膜细胞数分别为(52.67±2.73)、(117.30±3.48)和(62.00±3.06)。结论 HAI-1 m RNA表达水平与子宫内膜癌细胞侵袭迁移能力呈负相关。对HAI-1抑癌机制的进一步研究,将有望使HAI-1成为今后子宫内膜癌诊断及治疗中的新靶点。展开更多
The integral membrane,Kunitz-type serine protease inhibitors HAI-1 and HAI-2,can suppress the proteolytic activity of the type 2 transmembrane serine protease matriptase with high specificity and potency.High levels o...The integral membrane,Kunitz-type serine protease inhibitors HAI-1 and HAI-2,can suppress the proteolytic activity of the type 2 transmembrane serine protease matriptase with high specificity and potency.High levels of extracellular matriptase proteolytic activity have,however,been observed in some neoplastic B-cells with high levels of endogenous HAI-2,indicating that HAI-2 may be an ineffective matriptase inhibitor at the cellular level.The different effectiveness of the HAIs in the control of extracellular matriptase proteolytic activity is examined here.Upon inducing matriptase zymogen activation in the HAI Teton Daudi Burkitt lymphoma cells,which naturally express matriptase with very low levels of HAI-2 and no HAI-1,nascent active matriptase was rapidly inhibited or shed as an enzymatically active enzyme.With increasing HAI-1 expression,cellular matriptase-HAI-1 complex increased,and extracellular active matriptase decreased proportionally.Increasing HAI-2 expression,however,resulted in cellular matriptase-HAI-2 complex levels reaching a plateau,while extracellular active matriptase remained high.In contrast to this differential effect,both HAI-1 and HAI-2,even at very low levels,were shown to promote the expression and cell-surface translocation of endogenous matriptase.The difference in the suppression of extracellular active matriptase by the two closely related serine protease inhibitors could result from the primarily cell surface expression of HAI-1 compared to the mainly intracellular localization of HAI-2.The HAIs,therefore,resemble one another with respect to promoting matriptase expression and surface translocation but differ in their effectiveness in the control of extracellular matriptase enzymatic activity.展开更多
文摘目的探讨HAI-1表达水平与子宫内膜癌细胞株侵袭迁移能力的相关性。方法体外培养人子宫内膜癌细胞株,采用Real-time q PCR技术检测HEC-1A、HEC-1B和RL952三株子宫子宫内膜癌细胞中HAI-1 m RNA表达水平,应用细胞划痕实验及穿膜小室实验,观察三株子宫内膜癌细胞体外的侵袭及迁移能力。结果 (1)HAI-1 m RNA在三株细胞中的相对表达量分别为(0.3042±0.0101)、(0.0032±0.0001)和(0.2580±0.0096),HAI-1蛋白在HEC-1A、HEC-1B和RL952中均呈阳性表达,但在HEC-1B中表达低于HEC-1A及RL952。(2)细胞划痕实验结果显示,HEC-1A、HEC-1B和RL952三组迁移距离分别为(173.30±7.33)μm、(437.00±8.72)μm和(135.30±6.89)μm。(3)侵袭实验结果显示,HEC-1A、HEC-1B和RL952三组穿膜细胞数分别为(52.67±2.73)、(117.30±3.48)和(62.00±3.06)。结论 HAI-1 m RNA表达水平与子宫内膜癌细胞侵袭迁移能力呈负相关。对HAI-1抑癌机制的进一步研究,将有望使HAI-1成为今后子宫内膜癌诊断及治疗中的新靶点。
基金This study was supported by Cancer Institute Grant R01 CA 123223(to MDJ and CYL)Grant(MAB-108-079)from the Ministry of Defense Medical Affairs Bureau+2 种基金Grants(CMNDMC10705,CMNDMC10813)from Chi-Mei Medical Center(to J.-K.Wang)Grant(TSGH-E-109213-003)from Tri-Service General Hospital(to Y.-Y.Wu)Grants(108-2311-B-016-001-,109-2320-B-016-004-)from Ministry of Science and Technology(to Y.-L.Chiu).
文摘The integral membrane,Kunitz-type serine protease inhibitors HAI-1 and HAI-2,can suppress the proteolytic activity of the type 2 transmembrane serine protease matriptase with high specificity and potency.High levels of extracellular matriptase proteolytic activity have,however,been observed in some neoplastic B-cells with high levels of endogenous HAI-2,indicating that HAI-2 may be an ineffective matriptase inhibitor at the cellular level.The different effectiveness of the HAIs in the control of extracellular matriptase proteolytic activity is examined here.Upon inducing matriptase zymogen activation in the HAI Teton Daudi Burkitt lymphoma cells,which naturally express matriptase with very low levels of HAI-2 and no HAI-1,nascent active matriptase was rapidly inhibited or shed as an enzymatically active enzyme.With increasing HAI-1 expression,cellular matriptase-HAI-1 complex increased,and extracellular active matriptase decreased proportionally.Increasing HAI-2 expression,however,resulted in cellular matriptase-HAI-2 complex levels reaching a plateau,while extracellular active matriptase remained high.In contrast to this differential effect,both HAI-1 and HAI-2,even at very low levels,were shown to promote the expression and cell-surface translocation of endogenous matriptase.The difference in the suppression of extracellular active matriptase by the two closely related serine protease inhibitors could result from the primarily cell surface expression of HAI-1 compared to the mainly intracellular localization of HAI-2.The HAIs,therefore,resemble one another with respect to promoting matriptase expression and surface translocation but differ in their effectiveness in the control of extracellular matriptase enzymatic activity.
