lncRNA SNHG16靶向PAR1调控肺癌发生、发展的机制研究  

Mechanism of lncRNA SNHG16 targeting PAR1 to regulate the occurrence and development of lung cancer

作　　者：李燕 刘杰 耿良 张璐 王歌 于小林 LI Yan;LIU Jie;GENG Liang;ZHANG Lu;WANG Ge;YU Xiaolin(Department of Integrated Chinese and Western Medicine,Affiliated Cancer Hospital of Zhengzhou University/Henan Provincial Cancer Hospital,Zhengzhou,Henan 450008,China)

机构地区：[1]郑州大学附属肿瘤医院/河南省肿瘤医院中西医结合科,郑州450008

出　　处：《重庆医学》2024年第10期1454-1459,1466,共7页Chongqing medicine

基　　金：国家自然科学基金项目(82204844,82102916);河南省科技攻关项目(222102310468)。

摘　　要：目的探讨长链非编码RNA(lncRNA)SNHG16调控PAR1对肺癌增殖、迁移、侵袭的影响及机制。方法收集2020年3月至2021年8月于该院手术切除的35例肺癌患者的肺癌组织及癌旁组织标本,同时培养肺癌细胞系(HCC827、A549、SK-LU-1、A427)和正常肺细胞系(MRC5)。利用表达载体pcDNA3.1构建生成PAR1过表达模型(pcDNA-PAR1),A549细胞转染后分为转染si-SNHG16、转染si-PAR1、转染si-SNHG16+pcDNA-PAR1及对照(转染si-NC)。采用实时荧光定量逆转录-PCR(qRT-PCR)检测肺癌细胞系(HCC827、A549、SK-LU-1、A427)、肺癌组织及癌旁组织标本中lncRNA SNHG16及PAR1表达水平,并验证各组细胞转染效率。MTT法和克隆形成测定各组细胞的增殖,流式细胞仪检测各组细胞凋亡,细胞划痕和Transwell实验检测各组细胞的迁移和侵袭能力,Western blot检测PAR1的蛋白表达变化。结果肺癌组织lncRNA SNHG16表达水平高于癌旁组织(P<0.05)。肺癌细胞系(HCC827、A549、SK-LU-1、A427)lncRNA SNHG16表达水平高于正常肺细胞系(MRC5),差异有统计学意义(P<0.05)。qRT-PCR结果显示,转染si-SNHG16后lncRNA SNHG16基因表达水平为对照的(21.02±0.04)%,转染si-PAR1后PAR1基因表达水平为对照的(19.06±0.02)%,pcDNA-PAR1的PAR1基因表达水平为对照的(2.70±0.00)倍,差异有统计学意义(P<0.05)。Western blot结果显示,各转染的PAR1蛋白表达水平与对照比较差异有统计学意义(P<0.05)。与对照比较,转染si-SNHG16的细胞活性更低,克隆形成、细胞迁移、侵袭能力明显受到抑制,细胞凋亡率更高(P<0.05),而pcDNA-PAR1可减弱转染si-SNHG16对细胞增殖、凋亡、迁移、侵袭的影响(P<0.05)。lncRNA SNHG16与PAR1表达水平呈正相关(r=0.61)。结论lncRNA SNHG16可通过靶向PAR1调控肺癌的发生、发展。Objective To investigate the effect and mechanism of long chain non-coding RNA(lncRNA)SNHG16 regulating PAR1 on the proliferation,migration and invasion of lung cancer.Methods From March 2020 to August 2021,lung cancer tissues and adjacent tissues of 35 patients with lung cancer were collected,and lung cancer cell lines(HCC827,A549,SK-LU-1,A427)and normal lung cell lines(MRC5)were simultaneously cultured.The overexpression model of PAR1(pcDNA-PAR1)was constructed by using the expression vector pcDNA3.1.A549 cells were divided into four groups after transfection:si-SNHG16,si-PAR1,si-SNHG16+pcDNA-PAR1 and control(transfection with si-NC).The expression levels of lncRNA SNHG16 and PAR1 in lung cancer cell lines(HCC827,A549,SK-LU-1,A427),lung cancer tissues and adjacent tissues were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction(qRT-PCR),and the transfection efficiency of each group was verified.MTT assay and clonal formation were used to determine the proliferation of cells in each group,flow cytometry was used to detect the apoptosis of cells in each group,cell scratch and Transwell test were used to detect the migration and invasion ability of cells in each group,and Western blot was used to detect the protein expression of PAR1.Results The expression level of lncRNA SNHG16 in lung cancer tissue was higher than that in adjacent tissues(P<0.05).The expression level of lncRNA SNHG16 in lung cancer cell lines(HCC827,A549,SK-LU-1,A427)was higher than that in normal lung cell lines(MRC5),with statistical significance(P<0.05).The results of qRT-PCR showed that the expression level of lncRNA SNHG16 gene was(21.02±0.04)%of the control after transfection of si-SNHG16,the expression level of PAR1 gene was(19.06±0.02)%of the control after transfection of si-PAR1,and the expression level of PAR1 gene was 2.70±0.00 folds of the control after transfection of pcDNA-PAR1,the difference was statistically significant(P<0.05).The results of Western blot showed that the expression l

关 键 词：肺癌 lncRNA SNHG16 PAR1 增殖 凋亡 迁移 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]