Biotech Research and Innovation Centre, University of Copenhagen, Denmark.
EMBO J. 2011 Sep 13;30(22):4628-41. doi: 10.1038/emboj.2011.331.
Autophagy is an evolutionarily conserved mechanism of cellular self-digestion in which proteins and organelles are degraded through delivery to lysosomes. Defects in this process are implicated in numerous human diseases including cancer. To further elucidate regulatory mechanisms of autophagy, we performed a functional screen in search of microRNAs (miRNAs), which regulate the autophagic flux in breast cancer cells. In this study, we identified the tumour suppressive miRNA, miR-101, as a potent inhibitor of basal, etoposide- and rapamycin-induced autophagy. Through transcriptome profiling, we identified three novel miR-101 targets, STMN1, RAB5A and ATG4D. siRNA-mediated depletion of these genes phenocopied the effect of miR-101 overexpression, demonstrating their importance in autophagy regulation. Importantly, overexpression of STMN1 could partially rescue cells from miR-101-mediated inhibition of autophagy, indicating a functional importance for this target. Finally, we show that miR-101-mediated inhibition of autophagy can sensitize breast cancer cells to 4-hydroxytamoxifen (4-OHT)-mediated cell death. Collectively, these data establish a novel link between two highly important and rapidly growing research fields and present a new role for miR-101 as a key regulator of autophagy.
自噬是一种细胞自我消化的进化保守机制,其中蛋白质和细胞器通过递送至溶酶体而降解。该过程的缺陷与包括癌症在内的许多人类疾病有关。为了进一步阐明自噬的调节机制,我们进行了功能筛选,以寻找调节乳腺癌细胞自噬通量的 microRNAs(miRNAs)。在这项研究中,我们确定了肿瘤抑制 miRNA,miR-101,作为基础、依托泊苷和雷帕霉素诱导的自噬的有效抑制剂。通过转录组谱分析,我们鉴定了三个新的 miR-101 靶标,STMN1、RAB5A 和 ATG4D。这些基因的 siRNA 介导的耗尽模拟了 miR-101 过表达的效果,表明它们在自噬调节中的重要性。重要的是,STMN1 的过表达可以部分挽救细胞免受 miR-101 介导的自噬抑制,表明该靶标具有功能重要性。最后,我们表明 miR-101 介导的自噬抑制可以使乳腺癌细胞对 4-羟基他莫昔芬(4-OHT)介导的细胞死亡敏感。总之,这些数据在两个非常重要和快速发展的研究领域之间建立了新的联系,并提出了 miR-101 作为自噬关键调节剂的新作用。