子宫颈鳞状细胞癌中超保守核糖核酸339的表达及其临床病理意义  

Expression and clinicopathological significance of ultra-conserved RNA-339 in cervical squamous cell carcinoma

作　　者：仇姝 陆红梅[2] 周洁 QIU Shu;LU Hongmei;ZHOU Jie(Department of Gynecology and Obstetrics,Yangzhou City Maternal and Child Health Care Hospital Affiliated to Yangzhou University,Yangzhou,Jiangsu,225000;Department of Pathology,Yangzhou City Maternal and Child Health Care Hospital Affiliated to Yangzhou University,Yangzhou,Jiangsu,225000;Department of Pathology,the Affiliated Hospital of Yangzhou University,Yangzhou,Jiangsu,225000)

机构地区：[1]扬州大学附属医院扬州市妇幼保健院妇产科,江苏扬州225000 [2]扬州大学附属医院扬州市妇幼保健院病理科,江苏扬州225000 [3]扬州大学附属医院病理科,江苏扬州225000

出　　处：《实用临床医药杂志》2023年第11期32-37,42,共7页Journal of Clinical Medicine in Practice

基　　金：江苏省卫生健康委科研项目(Z2021046);江苏省扬州市2021年市级计划-社会发展项目(YZ2021087)。

摘　　要：目的分析子宫颈鳞状细胞癌(CSCC)中超保守核糖核酸339(uc.339)的表达水平及其临床病理意义,探讨uc.339对宫颈癌细胞增殖的影响和相关机制。方法收集102例CSCC标本的临床特征。通过实时荧光定量聚合酶链反应(qRT-PCR)检测uc.339在CSCC组织中的表达水平,分析uc.339与临床病理特征的关系。通过MTT实验和克隆形成实验分析uc.339对细胞增殖的影响。采用生物信息学预测uc.339下游靶基因;采用qRT-PCR验证uc.339对靶基因微小RNA-339-5P(miR-339-5P)的影响;采用双荧光素酶报告实验验证uc.339对miR-339-5P的调控作用;采用回复实验证实uc.339对miR-339-5P调控及对宫颈癌细胞增殖的影响。结果qRT-PCR实验结果表明,CSCC组织中uc.339的表达水平高于癌旁正常宫颈组织,差异有统计学意义(P<0.05)。uc.339的表达与肿瘤大小(Pearson′s R=7.909)和病理分级(Pearson′s R=-4.859)显著相关(P<0.05),与患者年龄、人乳头瘤病毒(HPV)感染、淋巴结转移和TNM分期无相关性(P>0.05)。MTT实验提示过表达uc.339可促进宫颈癌细胞增殖,而降低uc.339表达则抑制了细胞的增殖,差异有统计学意义(P<0.05)。细胞克隆形成实验表明,过表达uc.339宫颈癌细胞克隆形成数目多于Scramble对照,降低uc.339表达宫颈癌细胞克隆形成数目低于si-对照,差异均有统计学意义(P<0.05)。生物信息学预测显示miR-339-5P是uc.339的靶基因;在宫颈癌细胞中过表达uc.339后,miR-339-5P表达水平下降,而降低uc.339表达后,miR-339-5P表达水平则升高,差异有统计学意义(P<0.05)。双荧光素酶报告实验证实uc.339与miR-339-5P特异性结合,并可负性调控miR-339-5P的表达。回复实验提示过表达miR-339-5P能消除uc.339对宫颈癌增殖的促进作用,而降低miR-339-5P的表达则能进一步激发宫颈癌细胞的增殖活性。结论CSCC中uc.339表达水平上调,可能是一个新的致癌基因。uc.339通过抑制miR-339-5P表达促进CSCC细胞的增殖�Objective To analyze the expression level and clinicopathological significance of ultra-conserved RNA-339(uc.339)in cervical squamous cell carcinoma(CSCC),and to explore the effect and related mechanism of uc.339 on proliferation of cervical canceRcells.Methods Clinical features of 102 CSCC specimens were collected.The expression level of uc.339 in the CSCC tissues was detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR),and the correlations between the uc.339 and the pathological characteristics were analyzed.MTT assay and clone formation assay were used to analyze the effect of uc.339 on proliferation of cells.Bioinformatics was use to predict the downstream target gene of uc.339;the qRT-PCRwas used to verify the effect of uc.339 on the target gene microRNA-339-5P(miR-339-5P);the double luciferase report experiment was used to verify the regulatory effect of uc.339 on miR-339-5P;the rescue reversion experiment was used to verify the effects of uc.339 on regulation of miR-339-5P and the proliferation of cervical canceRcells.Results The result of qRT-PCRshowed that the expression level of uc.339 in the CSCC tissues was significantly higheRthan that in the normal cervical tissues adjacent to canceR(P<0.05).The expression of uc.339 was significantly related to the tumoRsize(Pearson′s R=7.909)and pathological grading(Pearson′s R=-4.859)(P<0.05),but had no significant correlations with the patient′s age,human papillomavirus(HPV)infection,lymph node metastasis and TNM staging(P>0.05).MTT assay indicated that over-expression of uc.339 was able to promote the proliferation of cervical canceRcells,while decline in the expression of uc.339 was able to inhibit the proliferation of cells(P<0.05).The cell clone formation experiment showed that the numbeRof clone formation in cervical canceRcells overexpressing uc.339 was significantly higheRthan that in the Scramble controls,and the numbeRof clone formation in cervical canceRcells with reduced uc.339 expression was significantly loweRthan

关 键 词：子宫颈鳞状细胞癌 超保守核糖核酸339 临床特征 微小RNA-339-5P 细胞增殖 克隆形成实验 生物信息学 

分 类 号：R365[医药卫生—病理学] R711[医药卫生—基础医学]