PIWI蛋白相互作用RNA-47851在胃腺癌中的表达及其对增殖的影响  

Expression of PIWI-interacting RNA-47851 in gastric adenocarcinoma and its influence on proliferation

作　　者：朱锦丽 乔欣悦 严雪冰 王成海[2] ZHU Jinli;QIAO Xinyue;YAN Xuebing;WANG Chenghai(Department of Medical Record Statistics,the Affiliated Hospital of Yangzhou University,Yangzhou,Jiangsu,225009;Pathological Teaching and Research Office,Medical College of Yangzhou University,Yangzhou,Jiangsu,225009;Department of Oncology,the Affiliated Hospital of Yangzhou University,Yangzhou,Jiangsu,225009)

机构地区：[1]扬州大学附属医院病案统计科,江苏扬州225009 [2]扬州大学医学院病理教研室,江苏扬州225009 [3]扬州大学附属医院肿瘤科,江苏扬州225009

出　　处：《实用临床医药杂志》2024年第1期20-27,36,共9页Journal of Clinical Medicine in Practice

基　　金：江苏省卫生健康委科研项目(M2020024)。

摘　　要：目的探讨PIWI蛋白相互作用RNA-47851(piR-47851)在胃腺癌中的表达、临床病理意义及其对增殖的影响。方法通过实时荧光定量聚合酶链反应(qRT-PCR)检查79例胃腺癌组织中piR-47851的表达情况,分析piR-47851的表达水平与临床特征和生存预后的关系。通过细胞增殖实验观察piR-47851对胃癌细胞增殖活性的影响。应用信息学网站预测piR-47851的下游靶基因。建立MAPK1基因3′非翻译区(UTR)的野生型和突变型质粒,应用双荧光素酶报告系统验证piR-47851与MAPK1基因3′UTR结合从而抑制MAPK1蛋白合成。应用Rescue实验确认piR-47851对MAPK1直接调控作用,并探讨piR-47851/MAPK1对胃癌细胞增殖的影响。通过实验性裸鼠皮下荷瘤实验观察降低piR-47851表达对移植瘤体积和质量的影响;通过增殖指标Ki-67染色观察降低piR-47851表达对移植瘤细胞增殖活力的影响。结果qRT-PCR实验结果提示,79例胃腺癌组织中piR-47851的表达水平显著高于癌旁正常胃组织(P<0.05)。piR-47851表达水平与肿瘤大小、分化程度和生存预后密切相关(P<0.05)。生物信息学提示piR-47851与MAPK1基因3′UTR有互补的结合位点。双荧光素酶报告基因实验得出在MAPK1野生型转染组中,piR-47851能显著抑制荧光酶活性,而在MAPK1突变组中无抑制作用。CCK-8增殖活性实验表明,过表达piR-47851后,胃癌细胞增殖活性升高;降低piR-47851后,细胞增殖活性下降,差异均有统计学意义(P<0.05)。Rescue实验提示,在改变piR-47851和(或)MAPK1的表达后再通过qRT-PCR检测MAPK1的表达水平,MAPK1都能受到piR-47851的调控。在功能上过度表达MAPK1可以降低piR-47851对胃癌细胞的增殖促进作用,而减低MAPK1表达可进一步提升胃癌细胞的增殖活性。在裸鼠荷瘤实验中,降低piR-47851表达后肿瘤生长体积和质量显著减小(P<0.05)。增殖指数Ki-67染色表明减低piR-47851表达后增殖活性细胞数目显著低于对照组(P<0.05),�Objective To investigate the expression and clinical pathological significance of PIWI-interacting RNA-47851(piR-47851)in gastric adenocarcinoma and its influence on proliferation.Methods The expression of piR-47851 was detected in 79 gastric adenocarcinoma tissues by real time fluorescence quantitative polymerase chain reaction(qRT-PCR),and the correlation of piR-47851 expression level and clinical features with survival and prognosis were analyzed.The effect of piR-47851 on proliferation activity of gastric cancer cells was observed by cell proliferation experiments.Informatics websites were used to predict the downstream target genes of piR-47851.The wild-type and mutant plasmids for the 3'untranslated region(UTR)of MAPK1 gene were established,and a dual luciferase reporting system was used to verify that piR-47851 binded to the 3'UTR of MAPK1 gene,thereby inhibiting MAPK1 protein synthesis.The effect of overexpression/silencing of piR-47851 on MAPK1 expression level was examined through qRT-PCR experiment.Rescue experiment was used to confirm the direct regulatory effect of piR-47851 on MAPK1 and explore the effect of piR-47851/MAPK1 on the proliferation of gastric cancer cells.The effect of reduced piR-47851 expression on the volume and weight of transplanted tumors was observed in nude mice by xenograft tumor experiments;the effect of reduced piR-47851 expression on the cell proliferation activity of xenograft tumors was observed by Ki-67 staining.Results The qRT-PCR experiment result showed that the expression level of piR-47851 in 79 gastric adenocarcinoma tissues was significantly higher than that in normal gastric tissues adjacent to the cancer(P<0.05).The expression level of piR-47851 was significantly correlated with tumor size,degree of differentiation,and survival prognosis(P<0.05).Bioinformatics suggested that there was a complementary binding site between piR-47851 and MAPK1 gene 3'UTR.The dual luciferase reporter gene experiment showed that piR-47851 was able to significantly inhibit fluorescence

关 键 词：胃腺癌 PIWI蛋白相互作用RNA-47851 细胞增殖 增殖指数 生存预后 

分 类 号：R735.2[医药卫生—肿瘤] R446.11[医药卫生—临床医学] Q251[生物学—细胞生物学]