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本研究利用短发夹RNA(sh RNA)沉默SGC7901/ADM细胞MDR1基因表达,增强人胃癌SGC7901/ADM细胞对姜黄素的敏感性。根据MDR1基因序列设计3对编码sh RNA的DNA模板,克隆到p Silencer 3.1-H1 neo(p3.1)上构建3种sh RNA表达载体,转染SGC7901/ADM细胞,q RT-PCR和Western blotting检测MDR1基因沉默效果,用荧光显微镜观察细胞形态,MTT法检测细胞活力。结果显示,3种sh RNA表达载体转染细胞后均能不同程度沉默MDR1基因的表达,增强了细胞对姜黄素的敏感性。
In this study, short hairpin RNA (sh RNA) was used to silence MDR1 gene expression in SGC7901 / ADM cells and enhance the sensitivity of human gastric cancer cell line SGC7901 / ADM to curcumin. According to the MDR1 gene sequence, three pairs of DNA templates encoding sh RNA were designed and cloned into pSilencer 3.1-H1 neo (p3.1) to construct three kinds of sh RNA expression vectors and transfected into SGC7901 / ADM cells. Q RT-PCR and Western blotting The silencing effect of MDR1 gene was detected, cell morphology was observed by fluorescence microscopy, and cell viability was detected by MTT assay. The results showed that the three kinds of sh RNA expression vector transfected cells were able to silence the expression of MDR1 gene to varying degrees and enhance the sensitivity of cells to curcumin.