lncRNA ZNF649-AS1 Induces Trastuzumab Resistance by Promoting ATG5 Expression and Autophagy
lncRNA ZNF649-AS1 Induces Trastuzumab Resistance by Promoting ATG5 Expression and Autophagy
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DOI：
10.1016/j.ymthe.2020.07.019
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发表时间：
2020-11-04
影响因子：
12.4
通讯作者：
Dong, Huaying
中科院分区：
文献类型：
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作者：
Han, Mingli;Qian, Xueke;Dong, Huaying
迄今为止，长链非编码RNA（lncRNA）在曲妥珠单抗耐药中的调控机制尚未完全确定。在本研究中，我们鉴定了差异表达的lncRNA，并研究了其在乳腺癌曲妥珠单抗耐药中的调节作用。进行HiSeq测序和定量实时PCR以鉴定失调的lncRNA。通过质谱、RNA荧光原位杂交（RNA-FISH）和免疫沉淀分析来鉴定ZNF 649-AS 1与其他相关靶点（如多聚嘧啶束结合蛋白1（PTBP 1）和自噬相关蛋白5（ATG 5））之间的直接相互作用。我们的研究结果表明，ZNF 649-AS 1在曲妥珠单抗耐药细胞中的表达高于敏感细胞。ZNF 649-AS 1的表达增加与乳腺癌患者的反应较差和生存时间较短相关。在曲妥珠单抗处理的存在下，ZNF 649-AS 1通过H3 K27 ac修饰上调，并且ZNF 649-AS 1的敲低通过调节ATG 5表达和自噬逆转曲妥珠单抗抗性。在机制上，ZNF 649-AS 1与PTBP 1蛋白结合，进一步促进ATG 5基因的转录活性。总之，我们证明H3 K27 ac修饰诱导的ZNF 649-AS 1上调可能通过与PTBP 1相关并促进ATG 5转录而引起自噬和曲妥珠单抗抗性。
The regulatory mechanism of long non-coding RNAs (lncRNAs) in trastuzumab resistance is not well established to date. In this research, we identified differentially expressed lncRNA and investigated its regulatory role in trastuzumab resistance of breast cancer. HiSeq sequencing and quantitative real-time PCR were performed to identify the dysregulated lncRNAs. Mass spectrometry, RNA fluorescence in situ hybridization (RNA-FISH), and immunoprecipitation assays were performed to identify the direct interactions between ZNF649-AS1 and other associated targets, such as polypyrimidine tract binding protein 1 (PTBP1) and autophagy related 5 (ATG5). Our results showed that ZNF649-AS1 was more highly expressed in trastuzumab-resistant cells compared to sensitive cells. Increased expression of ZNF649-AS1 was associated with a poorer response and shorter survival time of breast cancer patients. ZNF649-AS1 was upregulated by H3K27ac modification at the presence of trastuzumab treatment, and knockdown of ZNF649-AS1 reversed trastuzumab resistance via modulating ATG5 expression and autophagy. Mechanically, ZNF649-AS1 was associated with PTBP1 protein, which further promoted the transcription activity of the ATG5 gene. In conclusion, we demonstrated that H3K27ac modification-induced upregulation of ZNF649-AS1 could cause autophagy and trastuzumab resistance through associating with PTBP1 and promoting ATG5 transcription.