Department of Urinary Surgery, The Affiliated Hospital of Chengde Medical University, Chengde, China.
Anticancer Drugs. 2024 Apr 1;35(4):362-370. doi: 10.1097/CAD.0000000000001571. Epub 2024 Feb 23.
To study the diagnostic value of mRNA expression in urinary exocrine body in bladder cancer.
From February 2022 to December 2022, 60 patients diagnosed with bladder cancer by pathology in the Department of Urology, Affiliated Hospital of Chengde Medical University were selected as the case group. In total, 40 healthy subjects receiving physical examinations were selected as the control group. 100 mL of morning urine samples were collected from the subjects in both groups based on the same standard. Three subjects were randomly selected from each group. Urinary exosomes were extracted by differential ultracentrifugation. High-throughput sequencing (RNA-seq) was used to detect mRNA expression profiles in urinary exosomes and identify differentially expressed genes. Bioinformatic analysis was performed to predict major biological functions of differentially expressed genes and related signaling pathways. RT-PCR validated expression levels of differentially expressed genes in urinary exosomes between the two groups. ROC curves evaluated the diagnostic value of differential genes for bladder cancer. Spearman's correlation analysis determined correlations between differentially expressed genes and the occurrence of bladder cancer. ROC curves speculated the diagnostic value of using combined differentially expressed genes.
Compared with normal subjects, there were 189 significantly differentially expressed genes in urinary exosomes of bladder cancer patients, including 33 up-regulated and 156 down-regulated. According to go and kyoto encyclopedia of genes and genomes (KEGG) analysis, the above differentially expressed genes may participate in the occurrence and development of bladder cancer through the MAPK pathway, PPAP signaling pathway, PI3K Akt signaling pathway and Hippo signaling pathway, affect protein and lipid metabolism, RNase activity, polysaccharide synthesis, signal transduction and other biological processes, and participate in cell proliferation, death, movement and adhesion, as well as cell differentiation and signal transduction. RT-PCR verified that the expression of tmeff1, SDPR, ACBD7, SCG2 and COL6A2 in the two groups of samples was statistically significant ( P < 0.05). The ROC curve showed that the area under curve area under the curve of the five differential genes were 0.6934, 0.7746, 0.7239, 0.6396 and 0.6610, respectively. The sensitivity was 42.11%, 64.86%, 47.37%, 73.53% and 76.47%, and the specificity was 90%, 81.36%, 96.36%, 61.02% and 58.18%, respectively. Spearman correlation analysis showed that tmeff1, SDPR and acbd7 were associated with the occurrence of bladder cancer. The ROC curve of the combined diagnosis of the three and the two combined diagnoses suggested that the area under the curve of the combined diagnosis of SDPR and acbd7 was 0.7945, the sensitivity was 89.09%, and the specificity was 60.53%.
The gene expression profile in urinary exosomes of bladder cancer patients has changed significantly, and the differential genes may play an important biological role in the occurrence and development of bladder cancer. The combined detection of urinary exosome SDPR and ACBD7 has a certain diagnostic value for bladder cancer.
研究尿外泌体 mRNA 表达在膀胱癌诊断中的价值。
选取 2022 年 2 月至 2022 年 12 月在承德医学院附属医院泌尿外科经病理诊断为膀胱癌的 60 例患者作为病例组，选择同期体检的 40 例健康者作为对照组。两组均按相同标准采集晨起 100 ml 尿液样本。每组随机抽取 3 例，采用差速超速离心法提取尿外泌体。采用高通量测序（RNA-seq）检测尿外泌体中 mRNA 表达谱，鉴定差异表达基因。生物信息学分析预测差异表达基因的主要生物学功能及相关信号通路。采用 RT-PCR 验证两组尿外泌体中差异表达基因的表达水平。ROC 曲线评估差异基因对膀胱癌的诊断价值。Spearman 相关性分析确定差异基因与膀胱癌发生的相关性。ROC 曲线推测联合差异表达基因的诊断价值。
与正常受试者相比，膀胱癌患者尿外泌体中有 189 个明显差异表达基因，其中上调 33 个，下调 156 个。根据基因本体（GO）和京都基因与基因组百科全书（KEGG）分析，上述差异表达基因可能通过 MAPK 通路、PPAP 信号通路、PI3K-Akt 信号通路和 Hippo 信号通路参与膀胱癌的发生发展，影响蛋白质和脂质代谢、RNase 活性、多糖合成、信号转导等生物学过程，并参与细胞增殖、死亡、运动和黏附以及细胞分化和信号转导。RT-PCR 验证两组样本中 tmeff1、SDPR、ACBD7、SCG2 和 COL6A2 的表达有统计学意义（ P ＜0.05）。ROC 曲线显示，5 个差异基因的曲线下面积 AUC 分别为 0.6934、0.7746、0.7239、0.6396 和 0.6610，灵敏度分别为 42.11%、64.86%、47.37%、73.53%和 76.47%，特异性分别为 90%、81.36%、96.36%、61.02%和 58.18%。Spearman 相关性分析显示，tmeff1、SDPR 和 acbd7 与膀胱癌的发生有关。SDPR 和 acbd7 联合诊断的 ROC 曲线提示，SDPR 和 acbd7 联合诊断的曲线下面积为 0.7945，灵敏度为 89.09%，特异性为 60.53%。
膀胱癌患者尿外泌体中的基因表达谱发生了明显改变，差异基因可能在膀胱癌的发生发展中发挥重要的生物学作用。尿外泌体 SDPR 和 ACBD7 的联合检测对膀胱癌具有一定的诊断价值。
