丹酚酸A逆转肺癌多药耐药基因MDR1相关microRNA的筛选与鉴定

发布时间：2019-06-05 09:39

【摘要】：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。
[Abstract]:Human lung cancer cell line A549 was divided into normal control group and administration group. The expression of MDR1 was measured by real-time quantitative PCR. MicroRNA chip was used to analyze the expression profile of microRNA in the administration group and the control group. Targetscan was used to predict the up-regulation of miRNA related to multi-drug resistance gene MDR1 in microRNA and real-time quantitative PCR was used to verify the selected miRNA. To study the reversal of lung cancer multi-drug resistance gene MDR1-related microRNA. by Salvianolic acid A. The results showed that compared with the control group, the expression level of MDR1 in the administration group was significantly down-regulated, and the miRNA expression profile of human lung cancer cell line A549 cells was analyzed by microRNA microarray, and a total of 426differentially expressed miRNA; were screened out. Then the target gene of differentially expressed miRNA was predicted, and it was found that four up-regulated miRNA were associated with multi-drug resistance gene MDR1. Four miRNA were verified by real-time quantitative PCR, and the results were consistent with those of the chip. Therefore, it is suggested that Salvianolic acid A down-regulate the expression of multi-drug resistance gene MDR1 in lung cancer by affecting the expression of miRNA and then regulating the target gene, which provides an experimental basis for further elucidating the mechanism of reversal of multi-drug resistance by traditional Chinese medicine.
【作者单位】： 南京中医药大学基础医学院;
【基金】：国家自然科学基金项目(81503486,81273638) 江苏省中医药局科技重点项目(ZD201502) 江苏省高校自然科学研究项目(14KJD360004)
【分类号】：R285

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