摘要
目的:探讨糖醛酸代谢相关酶二羰基/L-木酮糖还原酶(dicarbonyl and L-xylulose reductase,DCXR)对人肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)细胞增殖、迁移的影响及其潜在机制,并检测DCXR表达水平与肾癌患者的临床相关性。方法:利用生物信息学检测DCXR在泛癌及肾癌中的表达情况。应用定量逆转录聚合酶链反应(quantitative real-time PCR,qRT-PCR)和蛋白质印迹法方法,检测2022年6月30日—2023年1月30日期间解放军总医院第一医学中心收集的30例新鲜ccRCC组织与配对的癌旁组织,同时检测人ccRCC细胞株786-O、ACHN及人胚肾细胞株293T中DCXR的表达情况。将DCXR过表达载体分别转染入786-O、ACHN细胞,利用qRT-PCR与蛋白印迹法检测过表达效率,CCK-8(cell counting kit-8)实验、克隆形成实验检测细胞增殖;划痕实验、Transwell实验检测细胞迁移。对肾癌组织芯片进行免疫组织化学染色及临床相关性分析。最后通过GSEA富集分析预测DCXR可能参与调节的信号通路。结果:生物信息学显示肾癌中DCXR显著下调。在mRNA及蛋白水平上,肿瘤组织DCXR表达低于配对的癌旁肾组织,差异有统计学意义(P<0.05)。肾癌细胞786-O、ACHN中DCXR表达较人胚肾细胞低。过表达DCXR组786-O、ACHN细胞增殖活力及细胞克隆率均低于空载对照组,差异有统计学意义(P<0.05)。过表达DCXR组786-O及ACHN细胞划痕愈合面积及Transwell细胞迁移数均低于空载对照组,差异有统计学意义(P<0.05)。通过对本中心ccRCC组织芯片染色及相关临床数据分析显示,DCXR表达情况与ccRCC预后相关(P<0.05)。结论:DCXR在ccRCC中表达下降,过表达DCXR可抑制ccRCC的细胞增殖、迁移能力,并可能作为预测患者预后判断的生物学标记物。
Objective:To investigate the role and potential mechanism of glucuronate metabolism-related enzyme dicarbonyl and L-xylulose reductase(DCXR)in the proliferation and migration of human clear cell renal cell carcinoma(ccRCC)cells,and to detect the clinical relevance of DCXR expression level in patients with renal cell carcinoma.Methods:Bioinformatics was used to detect the expression of DCXR in pan-cancer and renal cell carcinoma.Quantitative real-time PCR(qRT-PCR)and Western blotting were used to detect the expression of DCXR in ccRCC tissues of 30cases and paired adjacent tissues collected from First Medical Center of PLA General Hospital from June 30th,2022to January 30th,2023,as well as in human ccRCC cell lines 786-O,ACHN and human embryonic kidney cell line 293T.The overexpression vector of DCXR was transfected into 786-O and ACHN cells,and the overexpression efficiency was detected by qRT-PCR and Western blotting.Cell Counting Kit-8(CCK-8)experiment and colony formation experiment were used to detect cell proliferation.Wound healing exper-iment and Transwell experiment were used to detect cell migration.Immunohistochemical staining and clinical relevance analysis were performed on renal cell carcinoma tissue chips.Finally,GSEA enrichment analysis was used to predict the signaling pathway in which DCXR may be involved.Results:Bioinformatics showed that DCXR was significantly down-regulated in renal cell carcinoma.In mRNA and protein levels,the expression of DCXR in tumor tissues was lower than that in paired adjacent renal tissues,and the difference was statistically significant(P<0.05).The expressions of DCXR in renal cancer cells 786-O and ACHN were lower than those in human embryonic kidney cells.The proliferation activity and cell clone rate of 786-O and ACHN cells in the overexpression DCXR group were lower than those in the controlf3 empty vector control group,with statistically significant differences(P<0.05).The scratch healing area and Transwell cell migration number of 786-O and ACHN cells in the ov
作者
欧阳清
郑斌
巫胜攀
王集琛
王雷
李修彬
马鑫
OUYANG Qing;ZHENG Bin;WU Shengpan;WANG Jichen;WANG Lei;LI Xiubin;MA Xin(Medical School of Chinese PLA,Beijing,100853,China;Department of Urology,the Third Medical Centre,Chinese PLA General Hospital)
出处
《临床泌尿外科杂志》
CAS
2024年第4期314-320,共7页
Journal of Clinical Urology
基金
国家自然科学基金(No:81802804)
中国人民解放军总医院国家优秀青年科学基金培养基金(No:2020-YQPY-006)
解放军总医院青年自主创新科学基金成长项目(No:22QNCZ029)。
关键词
二羰基/L-木酮糖还原酶
肾透明细胞癌
增殖
迁移
dicarbonyl and L-xylulose reductase
clear cell renal cell carcinoma
proliferation
migration
