miR-21/RASA1/RAS轴在结肠癌中的作用及预测EGFR单抗疗效的研究

Biochip technology will be used to detect the profiles of RAS-GAPs in colorectal cancerous tissues ,normal colon tissues and cancerous cell lines of colon; colon cancer cell lines with KRAS wild-type will be selected as a cell model, and the expression levels of miR-21 and RASA1 will be upregulated/downregulated respectively;the changes of RASA1,RAS-GTP,t/pERK and t/pAKT expression levels in colon cancer cell lines with overexpressed or silenced miR-21 and RASA1 will be detected by qRT-PCR、Western-blotting and ICC, and the influences on the changes of proliferation, apoptosis,clonization in colon cell lines and tumorigenesis in vivo will be observed;furthermore drug resistance to cetubximab will be detected, and drug-resistant and non drug-resistant colon cancer cell lines and clinical tissue samples with KRAS wild-type will be collected,besides the changes of miR-21,RASA1 expression levels will be measured.the result will clarify that the axis of miR-21/RASA1/RAS regulates the signal pathway of RAS/RAF/ERK & RAS/PI3K/AKT,and it gets involved in the pathogenesis of colon cancer with KRAS wild-type and the mechanisms of drug-resistance to cetubximab.it may play some roles in finding new biomarkers for the personalized therapies of colon cancer and its efficacy prediction of EGFR momoclonal antibodies.

应用组织芯片技术检测RAS-GAPs在结肠癌组织和正常结肠组织、结肠癌细胞株中的表达谱；选择KRAS 基因野生型结肠癌细胞株为细胞模型，分别上调/下调细胞中miR-21、RASA1的表达；采用qRT-PCR、Western-blot及ICC等技术检测miR-21、RASA1过表达或沉默状态下结肠癌细胞中RASA1及RAS-GTP、t/pERK、t/pAKT表达水平变化，并观察对结肠癌细胞增殖、凋亡、克隆形成及体内成瘤性变化的影响；再进行EGFR单抗耐药试验，选择耐药和非耐药的KRAS野生型结肠癌细胞株和临床组织标本，检测miR-21、RASA1的表达水平变化。结果将阐明mi-R21/RASA1/RAS轴调控RAS/RAF/ERK和RAS/PI3K/AKT信号通路，参与KRAS野生型结肠癌的发病机制和EGFR单抗耐药机制，为临床结肠癌的个体化治疗及EGFR单抗疗效预测寻找到新的标志物。

应用组织芯片技术检测RAS-GAPs在结肠癌组织和正常结肠组织、结肠癌细胞株中的表达谱；选择KRAS 基因野生型结肠癌细胞株为细胞模型，分别上调/下调细胞中miR-21、RASA1的表达；采用qRT-PCR、Western-blot及ICC等技术检测miR-21、RASA1过表达或沉默状态下结肠癌细胞中RASA1及RAS-GTP、t/pERK、t/pAKT表达水平变化，并观察对结肠癌细胞增殖、凋亡、克隆形成及体内成瘤性变化的影响；再进行EGFR单抗耐药试验，选择耐药和非耐药的KRAS野生型结肠癌细胞株和临床组织标本，检测miR-21、RASA1的表达水平变化。结果将阐明miR-21/RASA1/RAS轴调控RAS/RAF/ERK和RAS/PI3K/AKT信号通路，参与KRAS野生型结肠癌的发病机制和EGFR单抗耐药机制，为临床结肠癌的个体化治疗及EGFR单抗疗效预测寻找到新的标志物。