【摘 要】
：
目的：过表达CRKI/Ⅱ及低表达CRKII对Hca-P细胞体外增殖、克隆形成、迁移、侵袭、黏附影响；明确p130cas/CRK/Dock180/Rac1通路中起主导作用CRK成员；研究肿瘤潜在制剂去整合素rAdibitor通过该通路的作用机制.方法：1.构建pcDNA3.1/V5-HisB-CRKI过表达载体,稳转Hca-P,获得CRKI稳定上调细胞株,考察CRKI过表达Hca-P增殖、迁移、侵袭
【机 构】
：
大连医科大学生化教研室 大连医科大学生物技术系,大连,116044
【出 处】
：
中国生物化学与分子生物学会2016年全国学术会议
论文部分内容阅读
目的：过表达CRKI/Ⅱ及低表达CRKII对Hca-P细胞体外增殖、克隆形成、迁移、侵袭、黏附影响；明确p130cas/CRK/Dock180/Rac1通路中起主导作用CRK成员；研究肿瘤潜在制剂去整合素rAdibitor通过该通路的作用机制.方法：1.构建pcDNA3.1/V5-HisB-CRKI过表达载体,稳转Hca-P,获得CRKI稳定上调细胞株,考察CRKI过表达Hca-P增殖、迁移、侵袭、黏附、生存活力影响；CRKI过表达对Hca-P体内足垫成瘤能力和淋巴结转移能力的影响.2.构建pGPU6/GFP/Neo-shRNA-CRKII及-NC表达载体,稳转至Hca-P细,获得CRKII表达稳定下调细胞株,考察CRKII敲降对Hca-P增殖、克隆形成、迁移、侵袭、黏附能力影响.3.构建pcDNA3.1/V5-HisB-CRKII真核过表达载体,瞬转至Hca-P,检测CRKII高表达对Hca-P增殖、克隆形成、迁移、侵袭影响.4.CRKI/Ⅱ过表达及CRKII低表达对p130cas/CRK/Dock180/Rac1信号通路影响.5.rAdinbitor对Hca-P毒性、凋亡、迁移、侵袭、黏附能力的影响及通过CRK/DOCK180/Rac1、MEK/ERK和PI3K/Akt/Mtor调控Hca-P恶性表型研究.结果：1.CRKI过表达促进Hca-P体外增殖、迁移、侵袭、淋巴结黏附能力和生存活力；2.CRKI过表达促进Hca-P体内成瘤和淋巴结转移；3.CRKII低表达抑制Hca-P增殖、克隆形成、迁移、侵袭和淋巴结黏附能力；4.CRKII高表达促Hca-P增殖、迁移及侵袭；5.CRKII表达不影响Hca-P中p130cas/CRK/Dock180/Rac1,CRKI表达变化对p130/casCRK/Dock180/Rac1影响明显；6.rAdinbitor抑制Hca-P增殖,促Hca-P凋亡；rAdinbitor通过CRK/DOCK180/Rac1、MEK/ERK和PI3K/Akt/Mtor通路显著抑制Hca-P迁移、侵袭和淋巴结黏附能力.结论：1.CRKI/Ⅱ表达水平与Hca-P恶性表型正相关；2.CRKI在调控 p130cas/CRK/Dock180/Rac1通路中起主要作用；3.rAdinbitor通过CRKI介导的信号通路来调控Hca-P恶性行为.
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