piR-128通过DNA甲基化酶3B对结直肠癌的调节作用及诊断价值  

Regulatory effect and diagnostic value of piR-128 on colorectal cancer through DNA methylase 3B

作　　者：高晓斌[1] 武雪亮[1] 赵轶峰[1] 聂双发[1] 梁峰[1] 张迎春[1] GAO Xiaobin;WU Xueliang;ZHAO Yifeng;NIE Shuangfa;LIANG Feng;ZHANG Yingchun(Department of General Surgery,the First Affiliated Hospital of Hebei North University,Hebei Province,Zhangjiakou075000,China)

机构地区：[1]河北北方学院附属第一医院普通外科,河北张家口075000

出　　处：《中国医药导报》2022年第28期11-16,32,共7页China Medical Herald

基　　金：河北省医学科学研究课题计划项目(20200515);河北省张家口市科技和地震局市级科技计划自筹经费项目(1521076D)。

摘　　要：目的检测piR-128通过DNA甲基化酶(DNMT)3B对结直肠癌的调节作用及其诊断价值。方法选取2018年2月至2020年1月于河北北方学院附属第一医院普通外科诊断为结直肠癌的54例患者的手术标本,采用qRT-PCR检测结直肠癌组织及邻近正常组织中piR-128和DNMT1、DNMT3A、DNMT3B的表达水平,通过Spearman相关分析评估piR-128和DNMT3B之间的关联,并进一步分析不同piR-128和DNMT3B mRNA表达水平患者与其临床特征的关系,通过受试者操作特征曲线评估piR-128在结直肠癌中的诊断和预后价值。选取人结直肠癌细胞LIM1215,将其分为对照组(转染空白抑制剂、si-空白)、piR-128抑制剂组(转染piR-128抑制剂)、si-DNMT3B组(转染si-DNMT3B)、抑制剂+Vector组(转染piR-128抑制剂+DNMT3B Vector),通过Western blot、qRT-PCR分析piR-128和DNMT3B的相互调控关系,通过transwell实验、细胞划痕实验和流式细胞术分析piR-128和DNMT3B对结直肠癌细胞侵袭、迁移和凋亡的作用。结果结直肠癌组织中piR-128、DNMT3BmRNA的表达水平高于邻近正常组织,差异有统计学意义(P<0.05)。相关性分析结果显示,结直肠癌组织中piR-128与DNMT3B mRNA表达水平呈正相关(rs>0,P<0.05)。piR-128抑制剂组、抑制剂+Vector组piR-128、DNMT3B mRNA水平低于对照组,si-DNMT3B组DNMT3B mRNA水平低于对照组,抑制剂+Vector组DNMT3B mRNA水平高于si-DNMT3B组,piR-128抑制剂组、si-DNMT3B组、抑制剂+Vector组DNMT3B蛋白水平均低于对照组,抑制剂+Vector组DNMT3B蛋白水平高于si-DNMT3B组,差异有统计学意义(P<0.05)。si-DNMT3B组、piR-128抑制剂组、抑制剂+Vector组在24、36、48、72 h的吸光度均低于对照组,差异有统计学意义(P<0.05)。piR-128抑制剂组、si-DNMT3B组、抑制剂+Vector组细胞侵袭、迁移率均低于对照组,凋亡率高于对照组,抑制剂+Vector组侵袭、迁移率高于si-DNMT3B组、piR-128抑制组,凋亡率低于si-DNMT3B组,差异有统计学意义(P<0.05)。结论pObjective To detect the regulatory effect and diagnostic value of piR-128 on colorectal cancer through DNA methylase(DNMT)3B.Methods A total of 54 patients diagnosed with colorectal cancer in the Department of General Surgery,the First Affiliated Hospital of Hebei North University from February 2018 to January 2020 were selected.The expression levels of piR-128 and DNMT1,DNMT3A,and DNMT3B in colorectal cancer tissues and adjacent normal tissues were detected by qRT-PCR.Spearman correlation analysis was used to evaluate the association between piR-128 and DNMT3B,and the relationship between patients with different levels of piR-128 and DNMT3B mRNA expression and their clinical characteristics was further analyzed.Receiver operation characteristic curve was used to evaluate the diagnostic and prognostic value of piR-128 in colorectal cancer.Human colorectal cancer cell line LIM1215 was selected and divided into control group(transfected with blank inhibitor and si-blank),piR-128 inhibitor group(transfected with piR-128 inhibitor),SI-DNMT3B group(transfected with si-DNMT3B)and inhibitor+Vector group(transfected with piR-128 inhibitor and DNMT3B Vector).Western blot and qRT-PCR were used to analyze the mutual regulatory relationship between piR-128 and DNMT3B.Transwell assay,cell scratch assay,and flow cytometry were used to analyze the effects of piR-128 and DNMT3B on the invasion,migration,and apoptosis of colorectal cancer cells.Results The levels of piR-128 and DNMT3B mRNA expression in colorectal cancer tissues were higher than those in adjacent normal tissues,and the differences were statistically significant(P<0.05).The results of correlation analysis showed that there was a positive correlation between the expression level of piR-128 and DNMT3BmRNA in colorectal cancer tissues(rs>0,P<0.05).The levels of piR-128 and DNMT3B mRNA in the piR-128 inhibitor group and inhibitor+Vector group were lower than those in the control group,the levels of DNMT3B mRNA in the si-DNMT3B group were lower than those in the contro

关 键 词：DNA甲基化酶3B piR-128 结直肠癌 作用机制 

分 类 号：R735.3[医药卫生—肿瘤]