摘要
目的通过高通量测序探索不同民族前列腺癌患者生物学差异的候选miRNA,寻找潜在的肿瘤标志物。方法选取2020年10月-2021年8月新疆医科大学第一附属医院收治的病理确诊为汉族(HANC组)、维吾尔族(WEIC组)前列腺癌患者各18例,哈萨克族(HAC组)前列腺癌患者3例,前列腺增生(HANN组)患者18例。使用穿刺组织提取RNA进行测序,筛选差异表达的miRNA并进行靶基因预测及功能分析。采用qRT-PCR技术检测5个候选miRNA表达水平并进行扩大样本量验证。结果通过高通量测序,HANC、WEIC、HAC、HANN各组分别获得18490709条、14440655条、17192254条、17069016条原始测序序列,经过对测序片段碱基的质量检验和长度筛选,各组获得干净序列分别为15814065条、13418490条、16318109条、16399958条。筛选的差异表达miRNA中WEIC组较HAC组上调4个miRNA,下调8个miRNA;WEIC组较HANC组上调15个miRNA,下调21个miRNA;HAC组较HANC组上调20个miRNA,下调16个miRNA;HANC组较HANN组上调69个miRNA,下调76个miRNA。功能预测显示,不同民族间差异miRNA的靶基因参与的KEGG通路具有一定的相似性;qRT-PCR验证了5个候选miRNA,结果显示HANC组和WEIC组中hsa-miR-4485-3p和hsa-miR-619-5p表达水平高于HANN组,hsa-miR-339-3p表达水平则低于HANN组(P<0.05);与HANC组相比,WEIC组中hsa-miR-1973、hsa-miR-4485-3p和hsa-miR-619-5p表达水平下调,hsa-miR-339-3p和hsa-miR-423-5p表达水平则上调,但两民族间各miRNA表达比较,差异无统计学意义(P>0.05)。此外,qRT-PCR验证结果与高通量结果表达趋势一致。结论hsa-miR-619-5p和hsa-miR-339-3p有潜力作为前列腺癌与前列腺增生患者分子诊断标志物。
Objective To explore the candidate miRNAs with biological differences in prostate cancer patients of different nationalities by high-throughput sequencing,and to find potential tumor markers.Methods Eighteen patients with prostate cancer(HANC group),18 patients with Uygur(WEIC group),3 patients with Kazak(HAC group)and 18 patients with benign prostatic hyperplasia(HANN group)were treated in the First Affiliated Hospital of Xinjiang Medical University from October 2020 to August 2021.RNA was extracted from punctured tissues for sequencing,and differentially expressed miRNAs were screened for target gene prediction and functional analysis.The expression levels of 5 candidate miRNA were detected by qRT-PCR technique and verified by enlarged sample size.Results Through high-throughput sequencing,an average of 18490709,14440655,17192254,and 17069016 original sequencing sequences were obtained from each group of HANC,WEIC,HAC,and HANN,respectively.After quality inspection and length screening of the sequenced fragments,15814065,13418490,16318109,and 16399958 clean sequences were obtained from each group,respectively.Among the differentially expressed miRNAs screened,compared with HAC group,4 miRNAs were up-regulated and 8 miRNAs were down-regulated in WEIC group;compared with HANC group,15 miRNAs were up-regulated and 21 miRNAs were down-regulated in WEIC group;compared with HANC group,20 miRNAs were up-regulated and 16 miRNAs were down-regulated in HAC group;compared with the HANN group,the HANC group up-regulated 69 miRNAs and down-regulated 76 miRNAs.Functional prediction showed that the KEGG pathway involved in the target genes of different miRNAs between different ethnic groups had certain similarities.Five candidate miRNAs were verified by qRT-PCR.The results showed that the expression levels of hsa-miR-4485-3p and hsa-miR-619-5p in HANC group and WEIC group were higher than those in HANN group,while the expression level of hsa-miR-339-3p was lower than that in HANN group(P<0.05).Compared with HANC group,the expression levels of hsa-miR-1973,hsa-miR-4485-3p and hsa-miR-619-5p in WEIC group were down-regulated,while the expression levels of hsa-miR-339-3p and hsa-miR-423-5p were up-regulated,but there was no significant difference in the expression of each miRNA between the two nationalities(P>0.05).In addition,the qRT-PCR results were consistent with the high-throughput results.Conclusion hsa-miR-619-5p and hsa-miR-339-3p have potential as molecular diagnostic markers for prostate cancer and benign prostatic hyperplasia.
作者
王文光
王玉杰
刘强
马军
李前进
木拉提·热夏提
阿斯木江·阿不拉
WANG Wen-guang;WANG Yu-jie;LIU Qiang;MA Jun;LI Qian-jin;Mulati Rexiati;Asimujiang Abula(Department of Urology,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang China)
出处
《医学信息》
2023年第1期7-16,共10页
Journal of Medical Information
基金
新疆维吾尔自治区自然科学基金(编号:2019D01C314)。
