摘要
目的:探究睾丸相关的高保守的致癌长链非编码RNA(THOR)在胃癌细胞中的表达及对胃癌细胞干性的影响。方法:收集徐州医科大学附属医院36对胃癌及癌旁正常组织标本,进行相关细胞培养、细胞活力测定、实时荧光定量PCR、慢病毒包和稳定的细胞株构建、mRNA稳定性分析、细胞球形形成分析、RNA测序分析、ALDH1活动分析,并对实验数据进行对比分析。结果:RNA测序分析发现胃癌组织中THOR水平较癌旁正常组织明显上调(P<0.01)。随着THOR的稳定敲除,胃癌细胞中的干性标志物(ALDH1,Nanog,Oct1,Oct 2,Oct 4,SOX2,SOX9,CD44)的表达显著降低(均P<0.01)。并且THOR的敲除降低了胃癌细胞形成球状体的能力,表现为球体尺寸变小和数量减少(P<0.01)。结论:THOR在胃癌细胞中的表达显著升高,敲除THOR可以降低胃癌细胞的干性。THOR与胃癌细胞中的干性标志物之间具有明显的作用关系,THOR可能是胃癌治疗的一个新的靶点。
Objective:To investigate the expression of testis-associated highly-conserved oncogenic long non-coding RNA(THOR)in gastric cancer cells and its influence on the stemness of gastric cancer cells.Methods:36 pairs of the specimens of gastric cancer tissue and normal tissues adjacent to cancer were collected from Affiliated Hospital of Xuzhou Medical University.Cell culture,cell viability test,real-time fluorescence quantitative PCR,construction of lentivirus envelope and stable cell line,analysis of mRNA stability,analysis of cell spheroid formation,RNA sequencing analysis,ALDH1 activity analysis were carried out,and the experimental data were compared and analyzed eventually.Results:RNA sequencing showed that the level of THOR in gastric cancer tissue was significantly higher than that in normal tissues(P<0.01).With the stable knockout of THOR,the expression of stemness markers including ALDH1,Nanog,Oct1,Oct 2,Oct 4,Sox2,Sox9 and CD44 in gastric cancer cells decreased significantly(P<0.01).In addition,the knockout of THOR decreased the ability of gastric cancer cells to form globules,which showed that the size and number of globules decreased(P<0.01).Conclusion:The expression of THOR in gastric cancer cells significantly increases,and the knockout of THOR can reduce the stemness of gastric cancer cells.There is a significant relationship between THOR and the stemness markers in gastric cancer cells,therefore THOR may be a new target for the treatment of gastric cancer.
作者
许雁晗
宋虎
宋军
宗启言
邹钰
李想
辛科
Xu Yanhan;Song Hu;Song Jun;Zong Qiyan;Zou Yu;Li Xiang;Xin Ke(School of Clinical Medicine,Xuzhou Medical University,Xuzhou,Jiangsu 221004,China;General Surgery,Affiliated Hospital of Xuzhou Medical University,Xuzhou,Jiangsu 221002,China;Institute of Digestive Diseases,Xuzhou Medical University,Xuzhou,Jiangsu 221002,China)
出处
《现代临床医学》
2020年第4期249-252,共4页
Journal of Modern Clinical Medicine
基金
基础医学国家级实验教学示范中心(徐州医科大学)资助项目(201810313065X)。
