肺鳞状细胞癌组织中LncRNA MIR205HG的表达及对癌细胞增殖克隆的影响和机制研究  

Expression of LncRNA MIR205HG in Squamous Cell Lung Carcinoma and Its Effect on Cell Proliferation and Cloning and Mechanism

作　　者：沈运涛 蔡笃雄[2] SHEN Yun-tao;CAI Du-Xiong(Department of Pediatrics,Qiongshan Maternal and Child Health Hospital of Haikou City,Haikou 571100,China;Department of Digestive Medicine Affiliated Hospital of Hainan Medical College,Haikou 570102,China)

机构地区：[1]海口市琼山区妇幼保健院儿科,海口571100 [2]海南医学院附属医院消化内科,海口570102

出　　处：《现代检验医学杂志》2022年第4期30-34,58,共6页Journal of Modern Laboratory Medicine

基　　金：海南省重点研发计划项目(ZDYF2018135)。

摘　　要：目的筛选肺鳞状细胞癌(squamous cell lung carcinoma,SQCLC)中差异表达且具有临床意义的潜在靶基因,并探究其在癌组织中的表达及其调控肺鳞状细胞癌发生发展的分子机制。方法通过临床数据库GEPIA找出在肺鳞状细胞癌中差异表达的长链非编码RNA(long noncoding RNAs,LncRNAs),进一步筛选出相较于正常肺组织表达差异性最显著的LncRNAs作为研究目的基因。通过PholyCSF数据库预测目的基因在肺鳞状细胞癌中发挥功能的形式。通过短发夹RNA(shRNA)介导敲低目的基因表达,利用细胞增殖实验、克隆形成实验验证目的基因在肺鳞状细胞癌发生发展中的重要性;通过分析与目的基因共表达基因参与的信号通路,探索其在肺鳞状细胞癌中发挥功能的潜在分子机制。结果研究最终选取在肺鳞状细胞癌中显著差异高表达的MIR205HG作为目的基因进行实验探究。肺鳞状细胞癌组织中MIR205HG表达显著高于正常肺组织(6.785±0.462 vs 1.084±0.036),差异有统计学意义(t=188.873,P<0.05),且随着临床分期越高MIR205HG表达水平越高(P=0.0472)。MIR205HG主要通过LncRNA形式在肺鳞状细胞癌中发挥功能。与shCTL组(0.988±0.039)相比,shMIR205HG#1组(0.709±0.032)和shMIR205HG#2组(0.736±0.026)细胞的增殖率明显降低,差异有统计学意义(F=66.163,P<0.001)。shMIR205HG#1组(0.324±0.021)和shMIR205HG#2组(0.402±0.023)细胞克隆形成速率明显低于shCTL对照组(1.809±0.019),差异有统计学意义(F=423.093,P<0.001)。shMIR205HG#1组(5.988±0.321)和shMIR205HG#2组(5.702±0.345)细胞中P53蛋白表达明显高于shCTL对照组(1.022±0.152),差异有统计学意义(F=285.386,P<0.001)。shMIR205HG#1组(3.857±0.362)和shMIR205HG#2组(3.698±0.314)细胞中下游P21蛋白表达亦明显高于shCTL对照组(1.106±0.253),差异有统计学意义(F=73.106,P<0.001)。结论lncRNA MIR205HG在肺鳞状细胞癌中显著高表达,可能通过P53信号通路参与肺鳞状细胞癌的增殖�Objective To screen potential target genes with differential expression and clinical significance in squamous cell lung carcinoma(SQCLC),and explore their expression in cancer tissues and the molecular mechanism of regulation of the occurrence and development of SQCLC.Methods Long non coding RNAs(LncRNAs)differentially expressed in SQCLC were identified through the clinical database GEPIA,and the lncRNAs with the most significant difference in expression compared with normal lung tissue were further selected as the target genes.The PholyCSF database was used to predict the functional pattern of the target gene in SQCLC.ShRNA mediated knockdown of target gene expression was used to verify the importance of target gene in the development and development of SQCLC by cell proliferation assay and clone formation assay.By analyzing the signaling pathways involved in the co-expression genes of the target genes,the potential molecular mechanisms of their function in SQCLC were explored.Results Finally,MIR205HG,which was significantly differentially expressed in SQCLC,was selected as the target gene for experimental exploration.The expression of MIR205HG in SQCLC was significantly higher than that in normal lung tissue(6.785±0.462 vs 1.084±0.036),the difference was statistically significant(t=188.873,P<0.05),and the higher the clinical stage was,the higher the MIR205HG expression level was(P=0.0472).MIR205HG plays a role in SQCLC mainly through LncRNA form.Compared with the shCTL group(0.988±0.039),the proliferation rate of shMIR205HG#1 group(0.709±0.032)and shMIR205HG#2 group(0.736±0.026)was significantly decreased,the difference was statistically significant(F=66.163,P<0.001).The rate of cell cloning formation in shMIR205HG#1 group(0.324±0.021)and shMIR205HG#2 group(0.402±0.023)was significantly lower than that in shCTL control group(1.809±0.019),the difference was statistically significant(F=423.093,P<0.001).The expression of P53 protein in shMIR205HG#1 group(5.988±0.321)and shMIR205HG#2 group(5.702±0.345)was

关 键 词：长链非编码核糖核酸(LncRNAs) MIR205HG 肺鳞状细胞癌 

分 类 号：R734.2[医药卫生—肿瘤] R730.43[医药卫生—临床医学]