单细胞测序分析显示 NEAT1 是前列腺癌的一个潜在生物标志物和治疗靶点。

Single-cell sequencing analysis revealed that NEAT1 was a potential biomarker and therapeutic target of prostate cancer.

机构信息

Department of Urology, Ningbo Medical Center LiHuiLi Hospital, Ningbo, Zhejiang Province, 315100, China.

出版信息

BMC Cancer. 2024 Oct 8;24(1):1242. doi: 10.1186/s12885-024-12926-y.

DOI:10.1186/s12885-024-12926-y

PMID:39379919

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11462789/

Abstract

BACKGROUND

Prostate cancer (PCa) usually manifests atypical symptoms in the early stage, and once symptoms appear, most PCa patients have developed to the advanced stage, failing to undergo radical surgery. In this study, PCa occurrence-related biomarkers were explored based on single-cell RNA sequencing (scRNA-seq) data.

METHODS

scRNA-seq data of prostate normal (Normal), benign prostatic hyperplasia (BPH), and PCa (Tumor) samples were acquired from the Gene Expression Omnibus (GEO). Cellular subsets associated with PCa occurrence were obtained using cell annotation. Additionally, the mRNA expression of nuclear enriched abundant transcript 1 (NEAT1) was detected by quantitative real-time PCR (qRT-PCR). The effects of NEAT1 on cell proliferation and apoptosis were analyzed by 5-ethynyl-2-deoxyuridine (EdU) and flow cytometry. Subsequently, cell-derived xenograft (CDX) models were constructed and divided into the LV-NC and LV-shNEAT1 groups. After the tumor tissues of CDX model mice in each group were extracted, the cell growth and Ki67 expression were observed separately using hematoxylin-eosin (H&E) staining and immunohistochemistry (IHC).

RESULTS

Ten cellular subsets were obtained via cell annotation, and significantly differential changes were observed between Basal intermediate and Luminal during the course of BPH to PCa. NEAT1Luminal was highly recruited in the Tumor group with low stemness and high malignancy scores. Matrix metallopeptidase 7 (MMP7) keratin 17 (KRT17)Basal intermediate had high ratios in the Tumor group with low stemness and high malignancy scores. The results of pseudotime analysis revealed that NEAT1Luminal in the Tumor group were consistently distributed with tumor stage cells. In vitro assays showed that NEAT1 expression was elevated in PCa cells, and NEAT1 knockdown could inhibit cell proliferation and induce apoptosis. CDX assays indicated that silencing NEAT1 could reduce the growth rate of PCa tumor volume in CDX model mice. H&E staining results showed that nuclei of tumor cells were reduced and exhibited lighter color in the LV-shNEAT1 group compared with the LV-NC group. IHC results showed that Ki67 positivity was significantly lower in the LV-shNEAT1 group than in the LV-NC group.

CONCLUSION

NEAT1 expression is increased in PCa, and NEAT1 can be a potential biomarker and therapeutic target for PCa.

摘要

背景

前列腺癌（PCa）在早期通常表现出非典型症状，一旦出现症状，大多数 PCa 患者已经发展到晚期，无法进行根治性手术。本研究基于单细胞 RNA 测序（scRNA-seq）数据探讨了与 PCa 发生相关的生物标志物。

方法

从基因表达综合数据库（GEO）中获取前列腺正常（Normal）、良性前列腺增生（BPH）和 PCa（Tumor）样本的 scRNA-seq 数据。使用细胞注释获得与 PCa 发生相关的细胞亚群。此外，通过实时定量 PCR（qRT-PCR）检测核富集丰富转录物 1（NEAT1）的 mRNA 表达。通过 5-乙炔基-2-脱氧尿苷（EdU）和流式细胞术分析 NEAT1 对细胞增殖和凋亡的影响。随后，构建细胞衍生异种移植（CDX）模型，并将其分为 LV-NC 和 LV-shNEAT1 组。提取每组 CDX 模型小鼠的肿瘤组织后，分别通过苏木精-伊红（H&E）染色和免疫组织化学（IHC）观察细胞生长和 Ki67 表达。

结果

通过细胞注释获得了 10 个细胞亚群，在 BPH 到 PCa 的过程中观察到 Basal intermediate 和 Luminal 之间存在显著的差异变化。在 Tumor 组中，NEAT1Luminal 招募程度较高，干性和恶性评分较低。基质金属蛋白酶 7（MMP7）/角蛋白 17（KRT17）Basal intermediate 在 Tumor 组中的比例较高，干性和恶性评分较低。假时间分析结果表明，Tumor 组中的 NEAT1Luminal 与肿瘤阶段细胞一致分布。体外实验表明，PCa 细胞中 NEAT1 表达升高，敲低 NEAT1 可抑制细胞增殖并诱导细胞凋亡。CDX 实验表明，沉默 NEAT1 可降低 CDX 模型小鼠中 PCa 肿瘤体积的生长速度。H&E 染色结果显示，与 LV-NC 组相比，LV-shNEAT1 组肿瘤细胞的细胞核减少，颜色变浅。免疫组化结果表明，LV-shNEAT1 组 Ki67 阳性率明显低于 LV-NC 组。