27-hydroxycholesterol and DNA damage repair: implication in prostate cancer.
27-hydroxycholesterol and DNA damage repair: implication in prostate cancer.
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DOI：
10.3389/fonc.2023.1251297
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发表时间：
2023
影响因子：
4.7
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中科院分区：
文献类型：
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我们之前报道过，前列腺癌（PC）中的胆固醇稳态是由27-羟基胆固醇（27HC）调节的，而将胆固醇转化为27HC的酶CYP27A1在PC中经常丢失。我们观察到恢复CYP27A1/27HC轴抑制PC的生长。在本研究中，我们探讨了27hc介导的抗pc作用的机制。我们采用体外模型和人类转录组学数据来研究27HC在PC中的作用机制。LNCaP （AR+）和DU145 （AR-）细胞分别用27HC或载药处理。使用Affymetrix GeneChip™微阵列系统进行转录组分析。测定差异表达，使用GSEA软件对MSigDB的标志基因集进行基因集富集分析。在mRNA和蛋白水平上验证了关键变化。分析了来自6个数据集的人类PC转录组，以确定CYP27A1与DNA修复基因表达特征之间的相关性。通过彗星检测评估DNA损伤。转录组分析显示，27HC处理下调了LNCaP细胞中与DNA损伤修复相关的Hallmark通路，降低了FEN1和RAD51的表达，并通过下调参与同源重组调控的基因诱导了“BRCAness”。一致地，我们发现在人类PC数据集中，阉割敏感PC （CSPC）中较高的CYP27A1表达（即较高的细胞内27HC）与DNA修复基因特征表达降低之间存在相关性。然而，这种相关性在转移性去势抵抗性PC （mCRPC）中不太明显。27HC增加了PC细胞中DNA损伤修复标记的表达，尤其是在AR+细胞中，但在AR-细胞中没有一致的作用，而在非肿瘤性前列腺上皮细胞中表达降低。在测试临床意义时，我们注意到27HC处理增加了LNCaP细胞的DNA损伤。通过补充胆固醇，而不是雄激素，效果是可逆的。最后，与olaparib（一种PARP抑制剂）联合使用，我们显示了加性DNA损伤效应。这些结果表明，27HC诱导“BRCAness”，一种被认为增加对PARP抑制剂敏感性的功能状态，并导致DNA损伤增加，特别是在CSPC中。鉴于人们逐渐认识到有缺陷的DNA损伤修复可以驱动PC生长，未来的研究需要测试27HC是否对CSPC中的PARP抑制剂和DNA损伤剂产生合成致死性。
We previously reported that cholesterol homeostasis in prostate cancer (PC) is regulated by 27-hydroxycholesterol (27HC) and that CYP27A1, the enzyme that converts cholesterol to 27HC, is frequently lost in PCs. We observed that restoring the CYP27A1/27HC axis inhibited PC growth. In this study, we investigated the mechanism of 27HC-mediated anti-PC effects. We employed in vitro models and human transcriptomics data to investigate 27HC mechanism of action in PC. LNCaP (AR+) and DU145 (AR-) cells were treated with 27HC or vehicle. Transcriptome profiling was performed using the Affymetrix GeneChip™ microarray system. Differential expression was determined, and gene set enrichment analysis was done using the GSEA software with hallmark gene sets from MSigDB. Key changes were validated at mRNA and protein levels. Human PC transcriptomes from six datasets were analyzed to determine the correlation between CYP27A1 and DNA repair gene expression signatures. DNA damage was assessed via comet assays. Transcriptome analysis revealed 27HC treatment downregulated Hallmark pathways related to DNA damage repair, decreased expression of FEN1 and RAD51, and induced “BRCAness” by downregulating genes involved in homologous recombination regulation in LNCaP cells. Consistently, we found a correlation between higher CYP27A1 expression (i.e., higher intracellular 27HC) and decreased expression of DNA repair gene signatures in castration-sensitive PC (CSPC) in human PC datasets. However, such correlation was less clear in metastatic castration-resistant PC (mCRPC). 27HC increased expression of DNA damage repair markers in PC cells, notably in AR+ cells, but no consistent effects in AR- cells and decreased expression in non-neoplastic prostate epithelial cells. While testing the clinical implications of this, we noted that 27HC treatment increased DNA damage in LNCaP cells via comet assays. Effects were reversible by adding back cholesterol, but not androgens. Finally, in combination with olaparib, a PARP inhibitor, we showed additive DNA damage effects. These results suggest 27HC induces “BRCAness”, a functional state thought to increase sensitivity to PARP inhibitors, and leads to increased DNA damage, especially in CSPC. Given the emerging appreciation that defective DNA damage repair can drive PC growth, future studies are needed to test whether 27HC creates a synthetic lethality to PARP inhibitors and DNA damaging agents in CSPC.
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影响因子：
8.2
作者：
de la Maza, Maria Dolores Fenor;Chandran, Khobe;Bono, Johann S. de
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影响因子：
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通讯作者：
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作者：
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作者：
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通讯作者：
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