沉默MDR1基因逆转SGC7901/ADM细胞对紫杉醇的耐药性

Silencing MDR1 Gene Expression Reverses the Drug-resistance of SGC7901/ADM Cells to Taxol

本研究利用短发夹sh RNA(short hairpin RNA)沉默SGC7901/ADM细胞MDR1基因,从而逆转SGC7901/ADM细胞对紫杉醇的耐药性。根据MDR1基因序列设计并合成编码sh RNA的DNA模板,构建3种靶向MDR1基因重组干扰载体,稳定转染SGC7901/ADM细胞,q RT-PCR检测MDR1 m RNA表达水平,Western blotting检测MDR1蛋白表达水平,MTT法检测细胞对紫杉醇的敏感性。结果表明,成功构建了靶向MDR1基因的3种重组表达载体,分别转染SGC7901/ADM细胞后均能不同程度沉默MDR1基因的表达,其中p2.1-3对MDR1沉默效果最好,m RNA和蛋白的沉默效率分别为78.5%和45%。紫杉醇对细胞的IC50值由(3.147±0.494)μmol/L降至(0.714±0.059)μmol/L,逆转率达到(78.22±1.906)%。结果表明,靶向MDR1的干扰表达载体能够抑制MDR1基因的表达,从而逆转SGC7901/ADM细胞对紫杉醇的耐药性。

In this study, we used shRNA （short hairpin RNA） to silence MDR1 gene to reverse the drug resistance of SGC7901/ADM cells to taxol. We designed and synthesized DNA template encoding shRNA, constructed three targeting MDR1 gene recombinant interference vector, stably transfected SGC7901/ADM cells, used qRT-PCR to detect the mRNA expression level of MDR1, used Western blotting to detect the protein expression level of MDR1, and used MTT to detect the sensitivity of cells to taxol. The results showed that we successfully constructed three recombinant expression vectors which could silence the expression of MDR1 gene in different extent after transfecting SGC7901/ADM cells. Among them, p2.1-3 had the best silencing effect on the expression of MDR1 and the silencing efficiency of mRNA and protein were 78.5% and 45% respectively. The IC50 value of cells of transfected p2.1-3 to taxol reduced from （3.147±0.494） μmol/L to （0.714±0.059） μmol/L and the reversal rate increased to （78.22± 1.906）%. The results showed that the targeting MDR1 of recombinant interference vectors could inhibit the expression of MDR1 gene and then reverse the drug resistance of SGC7901/ADM cells to taxol.