SPINK1慢病毒过表达载体和其稳转AsPC-1细胞株的构建及其对人胰腺癌细胞增殖的影响

来源 :第六届中国细胞生物学学会HIPPO年会暨大连医科大学2016年生命科学学术交流会 | 被引量 : 0次 | 上传用户:propelling
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  目的:本课题的目的是设计并制作人丝氨酸蛋白酶抑制因子Kazal 1型(SPINK1)基因的慢病毒过表达载体,并建立SPINK1过表达的稳转AsPC-1细胞株,探索其肿瘤生物学特性。方法:应用PCR方法扩增SPINK1基因序列,将其与慢病毒载体连接、转化,经电泳和基因测序进行鉴定,共转染293T细胞产生病毒颗粒,感染胰腺癌AsPC-1细胞,药筛感染后的阳性克隆并扩增,培养成SPINK1过表达稳转AsPC-1细胞株。通过Real-time PCR和Western blot的方法检测mRNA与蛋白的表达变化。用相关方法检测SPINK1基因对细胞增殖和克隆能力的影响。
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