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通过将人肺癌A549细胞分为正常对照组和给药组,采用实时定量PCR对MDR1的表达水平进行测定。运用microRNA芯片对给药组和对照组进行microRNA表达谱分析;采用Targetscan预测上调microRNA中与多药耐药基因MDR1相关的miRNA以及实时定量PCR的方法对筛选出的miRNA进行验证,研究丹酚酸A逆转肺癌多药耐药基因MDR1相关microRNA。实验结果表明,与对照组相比,给药组MDR1的表达水平明显下调;microRNA芯片对人肺癌A549细胞的给药组和对照组进行了miRNA表达谱分析,共筛出426个差异表达的miRNA;然后对差异表达上调的miRNA进行靶基因预测,发现有4个明显上调的miRNA与多药耐药基因MDR1相关。对4个miRNA进行实时定量PCR验证,结果与芯片一致。因此认为丹酚酸A下调肺癌多药耐药基因MDR1可能是通过影响miRNA表达进而调控靶基因,对进一步阐明中药逆转多药耐药作用机制提供了实验依据。
Human lung adenocarcinoma A549 cells were divided into normal control group and administration group, and the expression level of MDR1 was detected by real-time quantitative PCR. MicroRNA chip was used to analyze the microRNA expression profiles of the drug-treated group and the control group. Targetscan predict the up-regulated miRNAs associated with the multidrug resistance gene MDR1 in microRNAs and real-time quantitative PCR to validate the selected miRNAs, A reverses MDR1-associated microRNAs in lung cancer multidrug resistance genes. The results showed that compared with the control group, the expression level of MDR1 in the treated group was significantly decreased; microRNA chip miRNA expression profile analysis of the human lung cancer A549 cells administered group and the control group were screened out 426 differentially expressed miRNA Then, the miRNAs with differentially expressed genes were predicted to target genes and found that four miRNAs were significantly upregulated in multidrug resistance gene MDR1. The four miRNAs were verified by real-time PCR, and the results were consistent with the chip. Therefore, it is concluded that salvianolic acid A down-regulates the multidrug resistance gene MDR1 in lung cancer, which may regulate the target genes through affecting the miRNA expression, which provides experimental evidence for further elucidating the mechanism of multidrug resistance reversal in traditional Chinese medicine.