长链非编码RNA PTENP1通过miR-3611/PTEN基因途径调控宫颈癌进程的分子机制研究  

Study of molecular mechanism of regulation in cervical cancer progression by long non-coding RNA PTENP1 via miR-3611/PTEN gene pathway

作　　者：包利利 赵达 俞岩 周俏苗 BAO Lili;ZHAO Da;YU Yan;ZHOU Qiaomiao(The First Clinical Medical College,Hainan Medical College,Hainan Province,Haikou 570216,China;Department of Gynecology,Hainan Women and Children’s Medical Center,Hainan Province,Haikou 570100,China)

机构地区：[1]海南医学院第一临床学院,海南海口570216 [2]海南省妇女儿童医学中心妇科,海南海口570100

出　　处：《中国医药导报》2024年第3期19-27,共9页China Medical Herald

基　　金：海南省卫生健康行业科研项目(22A200105)。

摘　　要：目的 探讨长链非编码RNA PTENP1 (以下简称“PTENP1”)在宫颈癌细胞增殖、迁移和侵袭的分子机制。方法 选取2019年1月至2022年12月海南省妇女儿童医学中心诊断的54例的癌组织与癌旁组织,另选取宫颈癌细胞系(HeLa、SiHa、C33A、Caski)和正常宫颈上皮细胞系(H8),采用实时荧光定量PCR、蛋白质印迹法检测PTENP1、miR-3611、PTEN基因、PTEN蛋白水平。选取合适的宫颈癌细胞系,比较PTENP1在细胞质和细胞核中的表达情况,并进一步将其分为空白组(无处理)、pcDNA3.1组(转染pcDNA3.1)、pcDNA3.1-PTENP1组(转染pcDNA3.1-PTENP1)、NC组(阴性对照,转染模拟物或抑制剂NC)、miR-3611抑制剂组(转染miR-3611抑制剂)、pcDNA3.1-PTENP1+NC组(转染pcDNA3.1-PTENP1和NC)和pcDNA3.1-PTENP1+miR-3611 mimics组(转染pcDNA3.1-PTENP1和miR-3611模拟物)。比较空白组、pcDNA3.1组、pcDNA3.1-PTENP1组PTENP1、miR-3611、PTEN基因、PTEN蛋白及上皮间质转化的相关指标,采用细胞活力检测及流式细胞仪检测细胞增殖和凋亡情况;比较空白组、NC组、miR-3611抑制剂组miR-3611、PTEN基因、PTENP1水平;比较pcDNA3.1-PTENP1+NC组和pcDNA3.1-PTENP1+miR-361 1 mimics组的细胞增殖情况及上皮间质转化的相关指标。采用双萤光素酶报告基因及RNA下拉实验验证miR-3611与PTENP1、PTEN基因的靶向关系。结果 癌组织PTENP1、PTEN基因、PTEN蛋白水平低于癌旁组织,miR-3611水平高于癌旁组织(P<0.05)。HeLa、SiHa、C33A、Caski细胞PTENP1、PTEN基因、PTEN蛋白水平低于H8细胞,miR-3611水平高于H8细胞(P<0.05),后续实验选择HeLa、Caski细胞进行,PTENP1在HeLa、Caski细胞质中高表达。pcDNA3.1-PTENP1组PTENP1、凋亡率、上皮钙黏素、PTEN基因高于空白组,细胞增殖活力(培养48、72、96h)及ZEB1、Snail、波性蛋白、miR-3611低于空白组(P<0.05)。miR-3611抑制剂组miR-3611低于空白组,PTEN基因、PTENP1高于空白组(P<0.05)。pcDNA3.1-PTENP1+miR-361 1 mimics组细胞增殖�Objective To investigate the molecular mechanism of long non-coding RNA PTENP1(hereinafter referred to as "PTENP1") in the proliferation,migration,and invasion of cervical cancer cells.Methods Cancer tissues and adjacent tissues of 54 patients with cervical cancer diagnosed in Hainan Women and Children's Medical Center from January 2019 to December 2022 were selected,and cervical cancer cell lines(HeLa,SiHa,C33A,Caski) and normal cervical epithelial cell line(H8) were selected.The levels of PTENP1,miR-3611,PTEN gene,and PTEN protein were detected by real-time fluorescent quantitative PCR or Western blot.Appropriate cervical cancer cell lines were selected to compare the expression of PTENP1 in cytoplasm and nucleus,and further divided into blank group(no treatment),pcDNA3.1 group(transfected with pcDNA3.1),pcDNA3.1-PTENPl group(transfected with pcDNA3.1-PTENP1),NC group(negative control,transfected with eilther mimic or inhibitor,negative control),miR-3611 inhibitor group(transfected with miR-3611 inhibitor),pcDNA3.1-PTENP1+NC group(transfected with pcDNA3.1-PTENP1 and NG),and pcDNA3.1-PTENPl+miR-3611 mimics group(transfected with pcDNA3.1-PTENP1 and miR-3611 mimics).The levels of PTENP1,miR-3611,PTEN gene,PTEN protein,and epithelial mesenchymal transformation related indexes in blank group,pcDNA3.1 group,and pcDNA3.1-PTENP1 group were compared.Cell viability and flow cytometry were used to detect cell proliferation and apoptosis.The levels of miR-3611,PTEN gene,and PTENP1 in blank group,NC group,and miR-3611 inhibitor group were compared.The cell proliferation and epithelial mesenchymal transformation related indexes were compared between pcDNA3.1-PTENP1+NC group and pcDNA3.1-PTENPl+miR-3611 mimics group.Double luciferase reporter gene and RNA pull-down experiments were used to verify the targeting relationship between miR-3611 and PTENP1 and PTEN gene.Results The levels of PTENP1,PTEN gene,and PTEN protein in cancer tissues were lower than those in adjacent tissues,and the levels of miR-3611 were higher than th

关 键 词：长链非编码RNA 微RNA 竞争性结合机制 PTEN基因 宫颈癌 

分 类 号：R73-3[医药卫生—肿瘤]