苦参碱通过抑制microRNA-192调控顺铂耐受肺腺癌A549细胞对顺铂的敏感性

目的探讨苦参碱通过调控microRNA影响顺铂耐受A549(A549/DDP)细胞对顺铂的敏感性。方法采用生物信息学手段对GEO数据库中的相关microRNA芯片数据(访问编号GSE43249)进行数据挖掘,筛选出与A549细胞顺铂敏感性相关的microRNA。体外培养对顺铂敏感的A549细胞及A549/DDP,使用实时荧光定量PCR(qRT-PCR)法在体外培养的细胞中验证生物信息学的结果,并考察苦参碱对A549/DDP细胞中microRNA水平的影响。通过转染micro RNA-192(mi R-192)模拟序列上调A549/DDP细胞的mi R-192水平。CCK-8细胞活力实验考察苦参碱处理及(或)mi R-192上调对A549/DDP细胞顺铂敏感性的影响。结果芯片数据挖掘显示与顺铂敏感的A549细胞相比,A549/DDP细胞中共有11个microRNA存在显著变化(P0.05),其中mi R-192及mi R-194变化在体外培养的细胞中得到了验证。与未经处理的A549/DDP细胞相比,苦参碱处理后的A549/DDP细胞中mi R-192水平显著降低(P0.05)。苦参碱处理可以增加A549/DDP细胞对顺铂的敏感性,miR-192的上调可以消除这种效果。上调miR-192水平可诱导A549细胞的顺铂耐受。结论苦参碱可能通过抑制mi R-192调控A549对顺铂的敏感性。

Effect of matrine on sensitivity of A549/DDP cells to cisplatin by inhibiting microRNA-192

Objective To investigate the effect of matrine on increasing the sensitivity of A549/DDP cells to cisplatin by regulating microRNAs. Methods Data mining was performed on a microarray dataset (accession ID:GSE43249) from GEO database using bioinformatics analysis. Then microRNAs which related to the sensitivity of A549 cells to cisplatin were selected as candidate microRNAs. A549 and A549/DDP cells were cultured in vitro. The expression changes of candidate microRNAs were confirmed by qRT-PCR. The effect of matrine on the level of candidate microRNAs was also determined by qRT-PCR in A549/DDP cells. The level of microRNA-192 (miR-192) in A549/DDP cells was up-regulated by transfecting sequence of miR-192mimic. The CCK-8 assay was used to observe the effect of matrine and (or) miR-192 up-regulation on the sensitivity of A549/DDP cells to cisplatin. Results Data mining results showed that there were 11 microRNAs with significant changes in A549/DDP cells compared with cisplatin-sensitive A549 cells (P <0.05). The changes of miR-192 and miR-194 expression were confirmed in vitro. Compared with untreated A549/DDP cells, matrine treatment reduced the level of miR-192 in A549/DDP cells (P <0.05). Matrine treatment increased the sensitivity of A549/DDP cells to cisplatin. Up-regulation of miR-192 could antagonize this effect and induce the resistance of A549 cells to cisplatin. Conclusion Matrine may increase the sensitivity of A549 cells to cisplatin by inhibiting miR-192.