摘要
目的研究长链非编码RNA LINC01977对结直肠癌(CRC)及细胞系增殖的影响,分析与病理特征之间的关系,探讨LINC01977对结直肠癌的临床意义。方法收集2020年6月-2022年6月在郑州市中心医院首次确诊为CRC的50例患者的组织标本及临床病理资料,实时聚合酶链反应(qRT-PCR)检测CRC组织和正常组织中LINC01977的表达情况;检测LINC01977在结肠正常上皮细胞系NCM460和CRC细胞系Caco2、HCT8、HT29、SW620、HCT116、SW480中表达水平。分析LINC01977与患者临床病例特征之间的相关性。通过特异性siRNA敲低HCT116、SW480靶基因表达,将细胞分为对照组(si-NC组)和实验组(si-LINC01977组),检测敲低效率,采用细胞计数试剂盒-8(CCK-8)、克隆形成与EdU试验检测细胞增殖情况。结果LINC01977在CRC组织中表达为5.582(0.190,10.714),正常组织为2.151(0.022,6.076),差异有统计学意义(P<0.05)。LINC01977在HCT116中的表达量5.237(4.716,6.018)、SW480中的表达量7.246(6.287,7.826),均高于结肠上皮NCM460细胞表达量1.001(0.880,1.060),差异均有统计学意义(P均<0.05)。在癌症基因组图谱(TCGA)数据库中,LINC01977的表达量与CRC患者总生存期(OS)、无病生存期(DFS)及TNM分期相关(P<0.05);临床样本中LINC01977的表达量与肿瘤分化程度(χ^(2)=7.735,P=0.021)、TNM分期(χ^(2)=6.848,P=0.033)、淋巴结转移(χ^(2)=3.872,P=0.049)显著相关;CCK-8试验表明在培养48和72 h时,si-LINC01977(HCT116)组与si-NC(HCT116)组相比,细胞增殖差异均有统计学意义(t=11.424、20.699,P均<0.05);48和72 h时,si-LINC01977(SW480)组与si-NC(SW480)组相比,细胞增殖差异均有统计学意义(t=15.701、18.557,P均<0.05)。si-NC(HCT116)组克隆形成率显著高于si-LINC01977(HCT116)组(t=11.920),si-NC(SW480)组克隆形成率显著高于si-LINC01977(SW480)组(t=10.408),差异均有统计学意义(P均<0.05)。EdU实验表明si-NC(HCT116)组细胞数目显著高于si-LINC01977(HCT116)组(t=7.564),si-NC(SW480)组细胞数目显著高
Objective To investigate the effect of long non-coding RNA LINC01977 on colorectal cancer(CRC)and cell line proliferation;to analyze the relationship between LINC01977 and pathological features and explore the clinical significance of LINC01977 on colorectal cancer.Methods A total of 50 CRC tissue samples and clinicopathological data were collected from June 2020 to June 2022 in Zhengzhou Central Hospital for the detection of LINC01977 expression in CRC tissue and normal tissue by quantitative real-time polymerase chain reaction(qRT-PCR).The expression levels of LINC01977 in colon normal epithelial cell line NCM460 and CRC cell lines Caco2,HCT8,HT29,SW620,HCT116 and SW480 were detected.The correlation between LINC01977 and clinical characteristics of patients was analyzed.The cells were divided into control group(si-NC group)and experimental group(si-LINC01977 group)by knocking down the target gene expression of HCT116 and SW480 by specific siRNA,and the knockdown efficiency were tested.The cell proliferation was detected by cell counting kit-8(CCK-8),cloning and EdU assay.Results The relative expression level of LINC01977 in CRC tissue was 5.582(0.190,10.714),and that in normal tissue was 2.151(0.022,6.076),the difference was statistically significant(P<0.05).The relative expression level of LINC01977 in HCT116 was 5.237(4.716,6.018),and that of SW480 was 7.246(6.287,7.826),which were higher than that of colon epithelial NCM460 cells 1.001(0.880,1.060).The differences were statistically significant(all P<0.05).In the cancer genome atlas(TCGA)database,the expression of LINC01977 was related to overall survival(OS),disease free survival(DFS)and TNM stages of CRC patients(P<0.05).The expression of LINC01977 in clinical samples was significantly correlated with the degree of tumor differentiation(χ^(2)=7.735,P=0.021),TNM stage(χ^(2)=6.848,P=0.033)and lymph node metastasis(χ^(2)=3.872,P=0.049).CCK-8 test showed that there was difference in cell proliferation between si-LINC01977(HCT116)group and si-NC(HCT116)group
作者
邢孟宣
李璐
冯娇
张勇
褚菲菲
吴慧丽
XING Mengxuan;LI Lu;FENG Jiao;ZHANG Yong;CHU Feifei;WU Huili(Graduate School of Xinxiang Medical University,Xinxiang,Henan 453003,China;Department of Gastroenterology,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou,Henan 450000,China)
出处
《热带医学杂志》
CAS
2023年第5期607-612,584,共7页
Journal of Tropical Medicine
基金
河南省科技攻关计划项目(202102310382,212102310785)
