摘要
目的检测LncRNA TUG1在甲状腺癌组织及细胞中的表达,探究其对细胞增殖和迁移的影响。方法收集2015年1月~2018年6月在汉中市人民医院行手术治疗并经病理确诊的36例甲状腺癌患者癌组织标本及其相对应的正常癌旁甲状腺组织进行研究。采用qRT-PCR法检测甲状腺癌组织及SW1736,FTC-133细胞株中LncRNA TUG1表达水平。将shRNA插入慢病毒质粒中构建LncRNA TUG1低表达质粒(LV-shTUG1)。采用MTT法、集落形成实验及Transwell实验检测敲降LncRNA TUG1表达对甲状腺癌细胞增殖和迁移的影响;采用蛋白印迹分析实验检测LncRNA TUG1对EMT相关蛋白E-cadherin和N-cadherin表达的影响。结果与正常癌旁甲状腺组织相比,甲状腺癌组织中LncRNA TUG1高表达,差异有统计学意义(6.90±1.19 vs 1.51±1.02,t=20.634,P<0.000)。与Nthy-ori 3-1细胞相比,FTC-133细胞(1.00±0.09 vs 4.61±0.15)和SW1736细胞(1.00±0.09 vs 2.59±0.23)中LncRNA TUG1水平明显升高,差异有统计学意义(t=35.744,P<0.000;t=11.150,P<0.000)。敲降LncRNA TUG1表达后,SW1736细胞(1.31±0.19 vs 0.89±0.16)和FTC-133细胞(2.17±0.09 vs 1.68±0.05)增殖能力较对照组明显降低,克隆形成数目明显减少,差异有统计学意义(t=2.929,P=0.021;t=8.243,P=0.001)。敲降LncRNA TUG1表达后,FTC-133细胞迁移能力(1675.2±64.2 vs 898.1±156.4)较对照组显著降低,差异有统计学意义(t=7.961,P=0.001)。敲降LncRNA TUG1表达后,与对照组相比,E-cadherin表达量(0.74±0.06 vs 1.66±0.17)显著升高,N-cadherin表达量(1.27±0.18 vs 0.39±0.07)显著降低,差异有统计学意义(t=8.839,P<0.000;t=7.892,P=0.001)。结论LncRNA TUG1在甲状腺癌组织及细胞中高表达,促进甲状腺癌细胞增殖和迁移,可能通过促进EMT的形成进而促进甲状腺癌细胞的生物学行为。
Objective To detect the expression of LncRNA TUG1 in thyroid carcinoma tissues and cells,and explore its effect on cell proliferation and migration.Methods From January 2015 to June 2018,36 patients with thyroid cancer who underwent surgical treatment and were confirmed by pathology in Hanzhong People’s Hospital were collected,and their corresponding normal para-cancer thyroid tissues were collected for study.The expression levels of LncRNA TUG1 in thyroid carcinoma tissues and SW1736 and FTC-133 cell lines were detected by qRT-PCR.LncRNA TUG1 low expression plasmid(LV-ShTUG1)was constructed by inserting shRNA into lentivirus plasmid.MTT assay,colony formation assay and Transwell assay were used to detect the effects of knocked down LncRNA TUG1 expression on proliferation and migration of thyroid cancer cells.The effect of LncRNA TUG1 on the expression of EMT-related proteins E-cadherin and N-cadherin was detected by western blot analysis.Results The expression of LncRNA TUG1 in thyroid carcinoma tissues was significantly higher than that in normal paracarcinoma thyroid tissues(6.90±1.19 vs 1.51±1.02,t=20.634,P<0.000).LncRNA TUG1 levels in FTC-133 cells(1.00±0.09 vs 4.61±0.15)and SW1736 cells(1.00±0.09 vs 2.59±0.23)were significantly increased compared with Thy-ORI cells(t=35.744,P<0.000;t=11.150,P<0.000).After LncRNA TUG1 expression was knocked down,the proliferation ability of SW1736 cells(1.31±0.19 vs 0.89±0.16)and FTC-133 cells(2.17±0.09 vs 1.68±0.05)was significantly reduced compared with the control group,and the number of clones was significantly reduced,with statistically significant difference(t=2.929,P=0.021;t=8.243,P=0.001).After LncRNA TUG1 expression was knocked down,FTC-133 cell migration ability(1675.2±64.2 vs 898.1±156.4)was significantly lower than that of the control group,and the difference was statistically significant(t=7.961,P=0.001).After LncRNA TUG1 expression was knocked down,E-cadherin expression was significantly increased(0.74±0.06 vs 1.66±0.17)and N-cadherin expression was significantly decreased(1.27±0.18 vs 0.39±0.07)compared with the control group,the difference was statistically significant(t=8.839,P<0.000;t=7.892,P=0.001).Conclusion LncRNA TUG1 was highly expressed in thyroid cancer tissues and cells.Promoting the proliferation and migration of thyroid cancer cells may promote the biological behavior of thyroid cancer cells by promoting the formation of EMT.
作者
赵芳芳
郭红
陈嘉
ZHAO Fang-fang;GUO Hong;CHEN Jia(Department of Endocrinology,the People’s Hospital of Hanzhong City,Shaanxi Hanzhong 723000,China)
出处
《现代检验医学杂志》
CAS
2020年第6期42-47,共6页
Journal of Modern Laboratory Medicine
