miR-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为研究  

Investigate the Effect of miR-199a-3p on the Biological Behavior of NCI-H460 Lung Cancer Cells by Negatively Regulating CBX7

作　　者：孙党泽 丁超 朱昌生 张潍[2] 刘士源 马震川 李宇[2] SUN Dang-ze;DING Chao;ZHU Chang-sheng;ZHANG Wei;LIU Shi-yuan;MA Zhen-chuan;LI Yu(Department of thoracic surgery,Xi'an Chest Hospital,Xi'an,Shaanxi,710100,China;Department of thoracic surgery,The Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an,Shaanxi,710004,China)

机构地区：[1]西安市胸科医院胸外科,陕西西安710100 [2]西安交通大学第二附属医院胸外科,陕西西安710004

出　　处：《现代生物医学进展》2020年第3期455-459,496,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金青年科学基金项目(81602023)。

摘　　要：目的:探讨mi R-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为。方法:qRT-PCR法检测并比较肺癌组织、癌旁正常组织、肺癌细胞、正常肺上皮细胞中的mi R-199a-3p m RNA相对表达量。比较远处转移肺癌组织、未转移肺癌组织中mi R-199a-3p m RNA相对表达量。qRT-PCR法、Western Blot法检测并比较肺癌组织、癌旁正常组织中的CBX7 m RNA及蛋白的表达水平。荧光素酶活性法检测mi R-199a-3p与靶基因CBX7的结合。比较mi R-199a-3p模拟物转染组与阴性对照组的肺癌细胞中的CBX7 m RNA相对表达量及CBX7蛋白表达水平。CCK8实验检测mi R-199a-3p对肺癌细胞增殖的促进作用。Tranwell实验检测mi R-199a-3p对肺癌细胞侵袭与迁移能力的影响。结果:肺癌组织中mi R-199a-3p明显高于癌旁正常组织,发生远处转移的肺癌组织中mi R-199a-3p m RNA的表达量明显高于未发生转移的肺癌组织,差异有统计学意义(P<0.001)。肺癌组织中CBX7m RNA、CBX7蛋白表达水平均明显低于癌旁正常组织,差异有统计学意义(P<0.001)。荧光素酶活性法证实mi R-199a-3p可与靶基因CBX7结合抑制CBX7的表达。肺癌细胞中mi R-199a-3p m RNA的相对表达量明显高于正常肺上皮细胞,CBX7 m RNA相对表达量明显低于正常肺上皮细胞(P<0.05)。对于肺癌细胞,mi R-199a-3p模拟物转染组的CBX7 m RNA相对表达量及CBX7蛋白表达水平均明显低于阴性对照组(P<0.001)。CCK8实验证实mi R-199a-3p能够促进肺癌细胞的增殖,Tranwell实验证实mi R-199a-3p对肺癌细胞侵袭与迁移具有积极的促进作用。结论:mi R-199a-3p在肺癌的发生发展过程中发挥重要作用,能够通过抑制CBX7基因的表达,促进肺癌细胞的增殖、侵袭和转移。Objective:To explore the effect of mi R-199 a-3 p on the biological behavior of NCI-H460 lung cancer cells by negatively regulating CBX7.Methods:The relative expressions of mi R-199 a-3 p m RNA in lung cancer tissues,adjacent normal tissues,lung cancer cells and normal lung epithelial cells were detected and compared by qRT-PCR.The relative expression levels of mi R-199 a-3 p m RNA in distant metastatic lung cancer tissues and non-metastatic lung cancer tissues were compared.The expression levels of CBX7 m RNA and protein in lung cancer tissues and adjacent normal tissues were detected by qRT-PCR and Western Blot.The binding of mi R-199 a-3 p to target gene CBX7 was detected by luciferase activity.CBX7 m RNA expression levels and CBX7 protein expression levels in lung cancer cells transfected with mi R-199 a-3 p mimics and the negative control group were compared.CCK8 assay detected the promoting effect of mi R-199 a-3 p on the proliferation of lung cancer cells.Tranwell assay examined the effect of mi R-199 a-3 p on invasion and migration of lung cancer cells.Results:mi R-199 a-3 p in lung cancer tissues was significantly higher than that in adjacent normal tissues,and the expression level of mi R-199 a-3 p m RNA in lung cancer tissues with distant metastasis was significantly higher than that in lung cancer tissues without metastasis,the difference was statistically significant(P<0.001).The expression levels of CBX7 m RNA and CBX7 protein in lung cancer tissues were significantly lower than those in adjacent normal tissues(P<0.001).Luciferase activity confirmed that mi R-199 a-3 p could bind to target gene CBX7 and inhibit the expression of CBX7.The relative expression level of mi R-199 a-3 p m RNA in lung cancer cells was significantly higher than that in normal lung epithelial cells,and the relative expression level of CBX7 m RNA was significantly lower than that in normal lung epithelial cells(P<0.05).For lung cancer cells,the relative m RNA expression level and CBX7 protein expression level of mi R-199 a-3

关 键 词：肺癌 微小RNA miR-199a-3p CBX7 

分 类 号：R-33[医药卫生] R734.2