过表达lncRNA GAS5对甲状腺乳头状癌侵袭和迁移能力的影响及机制研究  

Effects of lncRNA GAS5 Overexpression on the Invasion and Migration Ability of Thyroid Papillary Carcinoma

作　　者：刘颖[1] LIU Ying(Department of Medicine,Liaoning Anshan Cancer Hospital,Anshan 114000)

机构地区：[1]辽宁省鞍山市肿瘤医院内科,辽宁鞍山114000

出　　处：《标记免疫分析与临床》2020年第10期1761-1766,1776,共7页Labeled Immunoassays and Clinical Medicine

基　　金：辽宁省科学技术计划项目(编号:2016030355)。

摘　　要：目的研究长链非编码RNA生长阻滞特异性转录本5(long non-coding RNA growth arrest-specific transcript 5,LncRNA GAS5)对甲状腺乳头状癌(papillary thyroid cancer,PTC)细胞侵袭和迁移能力的影响并探讨其作用机制。方法qRT-PCR检测LncRNA GAS5在PTC细胞系和正常甲状腺细胞系中的表达;选择低表达GAS5的PTC细胞进行过表达lncRNA GAS5慢病毒载体的转染,分为LV-NC组和LV-GAS5组。转染48 h后,Boyden实验检测各组细胞的侵袭能力,划痕实验检测各组细胞的迁移能力,Western-blot检测各组细胞中上皮间质转化(epithelial-mesenchymal transition,EMT)相关蛋白的表达;EMT诱导剂TGF-β处理细胞进行回复实验,分为LV-NC组、LV-GAS5组和LV-GAS5+TGF-β组,Boyden和划痕实验检测各组细胞侵袭和迁移能力变化。结果qRT-PCR结果显示LncRNA GAS5在PTC细胞系中的表达均低于在正常甲状腺细胞中的表达,B-CPAP细胞中GAS5的表达相对较低(P<0.05);转染过表达lncRNA GAS5慢病毒载体后,B-CPAP细胞侵袭和迁移能力降低(P<0.05),细胞中EMT相关蛋白N-cadherin、Vimentin表达降低和E-cadherin表达增加(P<0.05)。EMT诱导剂TGF-β处理后,与LV-NC组相比,LV-GAS5组和LV-GAS5+TGF-β组细胞侵袭和迁移能力降低,与LV-GAS5组相比,LV-GAS5+TGF-β组细胞侵袭和迁移能力增加(P<0.05)。结论GAS5在PTC细胞系中高表达,GAS5可能通过调控EMT抑制PTC的侵袭和转移能力。Objective To study the effect of long non-coding RNA growth arrest-specific transcript 5(LncRNA GAS5)on the invasion and migration ability of papillary thyroid cancer(PTC)and to explore its mechanism.Methods qRT-PCR was used to detect the expression of LncRNA GAS5 in PTC cell lines and normal thyroid cell lines.PTC cells with low expression of GAS5 were selected for transfection of lncRNA GAS5 lentiviral vectors,and were divided into the LV-NC group and LV-GAS5 group.48 hours after transfection,Boyden test was used to detect the invasion ability in each group of cells,wound test was used to detect the migration ability,and western-blot was used to detect the expression of epithelial-mesenchymal transition(EMT)-related proteins.EMT inducer TGF-βtreated cells were used for recovery experiments,which were divided into the LV-NC group,LV-GAS5 group and LV-GAS5+TGF-βgroup.Boyden and wound test were used to detect the cell invasion and migration ability of each group.Results The qRT-PCR results showed that the expression of LncRNA GAS5 in PTC cell lines was lower than that in normal thyroid cells,and the expression of GAS5 in B-CPAP cells was relatively low(P<0.05).After transfection with lentiviral vectors of over-expressed lncRNA GAS5,the invasion and migration ability of B-CPAP cells was reduced(P<0.05),also the expressions of N-cadherin,Vimentin were decreased in cells,and E-cadherin expression were increased(P<0.05).After treatment with the EMT inducer TGF-β,compared with the LV-NC group,the cell invasion and migration ability of the LV-GAS5 group and the LV-GAS5+TGF-βgroup were reduced.Compared with the LV-GAS5 group,the invasion and migration ability of LV-GAS5+TGF-βgroup were increased(P<0.05).Conclusion GAS5 is highly expressed in PTC cell lines,and GAS5 may inhibit the invasion and metastasis ability of PTC by regulating EMT level.

关 键 词：甲状腺乳头状癌 生长阻滞特异性转录本5 侵袭 迁移 

分 类 号：R736.1[医药卫生—肿瘤]