长链非编码RNA RP11-38P22对非小细胞肺癌生长和转移的作用及其机制  

Effects of long non-coding RNA RP11-38P22 on growth and metastasis of non-small cell lung cancer and possible mechanism

作　　者：王健[1] 文玉欣 丁培堃 任康奇 Wang Jian;Wen Yuxin;Ding Peikun;Ren Kangqi(Department of Thoracic Surgery,the Shengzhen People’s Hospital,the Second Clinical Medical College of Jinan University,Shenzhen 518020,China)

机构地区：[1]深圳市人民医院(暨南大学第二临床医学院)胸外科,518020

出　　处：《中华实验外科杂志》2020年第2期333-337,共5页Chinese Journal of Experimental Surgery

摘　　要：目的观察长链非编码RNA RP11-38P22在非小细胞肺癌(NSCLC)细胞株中表达水平的变化、对细胞生长和转移的影响以及调控的途径,探讨RP11-38P22在NSCLC的作用机制。方法2019年1月至2019年6月深圳市人民医院胸外科获取56对成对的NSCLC组织和相邻癌旁组织。通过实时定量聚合酶链反应(Real-time PCR)检测RP11-38P22在56例肺癌组织以及多个肺癌细胞株中的表达变化,并分析其表达水平的变化与NSCLC临床病理特征的相关性。将RP11-38P22过表达和敲低表达的A549和H1299细胞均设为实验组,将转染空载体病毒(即未干扰RP11-38P22表达的)的A549和H1299细胞设为对照组。改变RP11-38P22在细胞株A549和H1299的表达,通过细胞计数试剂盒(CCK-8)实验评估RP11-38P22对细胞生长的影响。通过流式细胞术、划痕实验及Transwell实验评估RP11-38P22对细胞凋亡、迁移和侵袭的影响。此外,通过免疫印迹技术探讨RP11-38P22发挥作用是否与p53通路相关。采用t检验比较两组的平均值,两组以上比较使用方差分析,使用Kaplan-Meier分析和对数秩检验比较生存数据。结果与癌旁组织相比较,RP11-38P22在肺癌组织中的表达明显降低(正常组织比腺癌组织为t=2.266,P<0.05;正常组织比鳞癌组织为t=2.313,P<0.05),且其表达的高低与肿瘤大小显著相关(χ^2=4.758,P<0.05)。与正常细胞(HBE)相比,5种NSCLC细胞系中,RP11-38P22表达也显著性下降,差异有统计学意义(A549细胞为F=45.674,P<0.01;H1299细胞为F=38.568,P<0.01;PC9细胞为F=23.337,P<0.05;H460细胞为F=18.459,P<0.05;Calu3细胞为F=22.102,P<0.05)。RP11-38P22过表达能够显著抑制A549和H1299细胞系的增殖能力[72 h A549细胞增殖率为对照组(110.275±5.663)%,实验组(65.735±3.706)%,F=16.342,P<0.05;72 h H1299细胞增殖率为对照组(125.357±6.733)%,实验组(76.857±3.935)%,F=11.360,P<0.05],并促进细胞的凋亡[A549细胞凋亡率为对照组(2.031±0.032)%,实验组(35.078±2.512)%,F=50.231,P<0Objective To investigate the mechanism of RP11-38P22 in non-small lung cancer cells(NSCLC)by studying the expression of RP11-38P22 in NSCLC cell lines,the effects on cell growth and metastasis,and the possible regulatory pathways.Methods Real-time quantitative polymerase chain reaction(Real-time PCR)was used to detect the expression changes of RP11-38P22 in 56 cases of lung cancer tissues and multiple lung cancer cell lines,and to analyze the correlation between the expression level changes and the clinicopathological characteristics of NSCLC.The expression of RP11-38P22 in A549 and H1299 cell lines was changed,and the effect of RP11-38P22 on cell growth was evaluated by cell counting kit-8(CCK-8)assay.The effects of RP11-38P22 on apoptosis,migration and invasion were evaluated by flow cytometry,scratch assay and Transwell assay,respectively.In addition,Western blotting was used to investigate whether the role of RP11-38P22 was related to the p53 pathway.Results The expression of RP11-38P22 in tumor tissues was significantly lower than that in adjacent tissues(normal tissues vs.adenocarcinoma:t=2.266,P<0.05;normal tissues vs.squamous cell carcinoma:t=2.313,P<0.05),and the expression level was correlated with tumor size(χ^2=4.758,P<0.05).RP11-38P22 expression was also significantly decreased in five NSCLC cell lines compared to normal cells(for A549 cell F=45.674,P<0.01;for H1299 cell F=38.568,P<0.01;for PC9 cell F=23.337,P<0.05;for H460 cell F=18.459,P<0.05;for Calu3 cell F=22.102,P<0.05).Overexpression of RP11-38P22 significantly inhibited proliferation of A549 and H1299 cell lines[at 72 h for A549 cell proliferation rate:control group(110.275±5.663)%vs.experimental group(65.735±3.706)%F=16.342,P<0.05;at 72 h for H1299 cell proliferation rate:control group(125.357±6.733)%vs.experimental group(76.857±3.935)%F=11.360,P<0.05]and promoted apoptosis[for A549 cell apoptosis rate:control group(2.031±0.032)%vs.experimental group(35.078±2.512)%,F=50.231,P<0.01;for H1299 cell apoptosis rate:control group(1.872±0.0

关 键 词：RP11-38P22 P53 非小细胞肺癌 生长 转移 

分 类 号：R734[医药卫生—肿瘤]