>
网站首页期刊介绍通知公告编 委 会投稿须知电子期刊广告合作联系我们
最新消息:
LncRNA DLG1-AS1调控miR-203/ZEB2轴诱导甲状腺乳头状癌细胞恶性化生长和转移的机制研究
作者:冯立新  翟传夫  谭清玉 
单位:临沂市中医医院 普外三科, 山东 临沂 276000
关键词:LncRNA DLG1-AS1 miR-203 上皮间质转化 甲状腺乳头状癌 增殖 转移 
分类号:R736.1
出版年·卷·期(页码):2021·40·第二期(133-141)
摘要:

目的:探讨LncRNA DLG1-AS1对甲状腺乳头状癌(PTC)细胞恶性化生长和转移的影响,并对其可能的分子机制进行研究。方法:体外培养PTC细胞(TPC1、KTC-1、B-CPAP、HTori-3)和正常甲状腺上皮细胞Nthy-ori3-1,采用实时荧光定量聚合酶链反应(qRT-PCR)检测细胞中DLG1-AS1和miR-203表达差异。将B-CPAP细胞分为空白对照组、阴性对照组(转染pcDNA3.1空载体和阴性对照序列)、shDLG1-AS1组(转染pcDNA3.1-shDLG1-AS1)、shDLG1-AS1+miR-203抑制物组(转染pcDNA3.1-shDLG1-AS1和miR-203抑制物)。通过CCK-8法和Transwell小室法检测细胞增殖、迁移和侵袭活性。蛋白质印迹法检测上皮间质转化(EMT)相关蛋白表达。双荧光素酶报告分析DLG1-AS1、miR-203、ZEB2之间的靶关系。结果:经qRT-PCR法检测,与Nthy-ori3-1细胞相比,TPC1、KTC-1、B-CPAP、HTori-3细胞中DLG1-AS1相对表达量均升高,同时miR-203相对表达量均降低(均P<0.05),选择DLG1-AS1相对表达量最高的B-CPAP细胞进行后续实验。与空白对照组和阴性对照组相比,shDLG1-AS1组细胞增殖、迁移、侵袭活性降低,ZEB2、N-cadherin、vimentin、β-catenin蛋白表达下调,同时E-cadherin表达上调(均P<0.05)。与shDLG1-AS1组相比,shDLG1-AS1+miR-203 inhibitor组细胞增殖、迁移、侵袭活性被逆转,ZEB2、N-cadherin、vimentin、β-catenin蛋白表达上调,同时E-cadherin表达下调(均P<0.05)。经双荧光素酶报告证实,DLG1-AS1可以直接调控miR-203,而ZEB2则是miR-203的下游靶基因。结论:LncRNA DLG1-AS1通过与miR-203竞争结合,消除miR-203对靶基因ZEB2表达的抑制,促进PTC细胞发生EMT,这可能是PTC恶性化生长和转移的重要机制。

Objective: To investigate the effect of LncRNA DLG1-AS1 on the proliferation and metastasis of papillary thyroid carcinoma(PTC) cells, and further to explore its mechanism. Methods: PTC cells TPC1,KTC-1,B-CPAP,HTori-3 and normal follicular cell Nthy-ori3-1 were used to detect DLG1-AS1 and miR-203 expressions by real time quantitative polymerase chain reaction(qRT-PCR) method. B-CPAP cells were divided into blank group, NC group(transfected pcDNA3.1-vector and negative control inhibitor), shDLG1-AS1 group(transfected pcdna 3.1-shDLG1-AS1), shDLG1-AS1+miR-203 inhibitor group(transfected pcdna3.1-shDLG1-AS1 and miR-203 inhibitor). The proliferation and invasion, migration activities of cells were detected by CCK-8 assay and transwell method. Western blotting was used to detect the relative expression of epithelial-mesenchymal transition(EMT) related proteins. The luciferase report was used to analyze the target relationship between DLG1-AS1, miR-203, and ZEB2. Results: qRT-PCR results showed compared with Nthy-ori3-1 cells, the relative expression of DLG1-AS1 in TPC1, KTC-1, B-CPAP and HTori-3 cells was increased, while the relative expression of miR-203 was decreased(P<0.05).B-CPAP cells with the highest relative expression of DLG1-AS1 were selected for follow-up study. Compared with blank group and NC group, the proliferation, migration and invasion of shDLG1-AS1 group cells were decreased, while the expression of ZEB2, N-cadherin, vimentin, β-catenin protein was down-regulated, and the expression of E-cadherin was up-regulated(P<0.05). Compared with shDLG1-AS1 group, shDLG1-AS1+miR-203 inhibitor group showed a reversal of cell proliferation, migration and invasion, while the expression of ZEB2, N-cadherin, vimentin, β-catenin protein was up-regulated and the expression of E-cadherin was down-regulated(P<0.05). DLG1-AS1 could directly regulate miR-203, and ZEB2 was the downstream target gene of miR-203. Conclusion: By combining with miR-203,LncRNA DLG1-AS1 eliminates the inhibition of miR-203 to ZEB2 expression, promotes the EMT of PTC cells, which may be an important mechanism of PTC malignant growth and metastasis.

参考文献:

[1] 谢洪,魏伯俊,申虹,等.甲状腺乳头状癌多原发癌的临床分析[J].中国现代医学杂志,2019,27(1):120-124.
[2] 程龙,邱伊波,许琳婉,等.PES1上调ERα/ERβ蛋白比率促进甲状腺乳头状癌细胞增殖和侵袭迁移[J].中山大学学报(医学科学版),2020,41(4):515-524.
[3] MENG Q,LI Z,PAN J,et al.Long noncoding RNA DUXAP8 regulates proliferation and apoptosis of ovarian cancer cells via targeting miR-590-5p[J].Hum Cell,2020,33(4):1240-1251.
[4] RUI X,XU Y,HUANG Y,et al.lncRNA DLG1-AS1 promotes cell proliferation by competitively binding with miR-107 and up-regulating ZHX1 expression in cervical cancer[J].Cell Physiol Biochem,2018,49(5):1792-1803.
[5] 潘钢,沈杰,孙思涵,等.MicroRNA-203靶向调控Bmi-1对甲状腺乳头状癌增殖及侵袭的影响[J].中华内分泌外科杂志,2018,14(4):269-273.
[6] 胡强,许斌,陈明,等.长链非编码RNA母系表达基因3在肿瘤中表达的研究现状[J].东南大学学报(医学版),2017,35(3):482-485.
[7] RANSOHOFF J D,WEI Y,KHAVARI P A.The functions and unique features of long intergenic non-coding RNA[J].Nat Rev Mol Cell Biol,2018,19(3):143-157.
[8] 朱岱阳,杜洋,范培芝,等.lncRNAs在甲状腺癌中的研究进展[J].实用医学杂志,2020,40(8):1116-1120.
[9] LI S.LncRNA DLG1-AS1 promotes cancer cell proliferation in triple negative breast cancer by downregulating miR-203[J].J Breast Cancer,2020,23(4):343-354.
[10] CHEN L,ZHOU Y,LI H.LncRNA,miRNA and lncRNA-miRNA interaction in viral infection[J].Virus Res,2018,257(9):25-32.
[11] DONG J,WANG Q,LI L,et al.Upregulation of long non-coding RNA small nucleolar RNA host gene 12 contributes to cell growth and invasion in cervical cancer by acting as a sponge for MiR-424-5p[J].Cell Physiol Biochem,2018,45(5):2086-2094.
[12] 戴磊,吴贤江,张舟径,等.miR-203表达异常与甲状腺乳头状癌发生发展的关系[J].现代实用医学,2018,25(7):854-856,858.
[13] 王媛,贺晶,王海英,等.血清CA125联合miR-31和miR-203在子宫内膜癌诊断和预后评估中的价值[J].现代医学,2019,44(11):1357-1359.
[14] 陈小波,豆倩影,黄立新,等.miR-203调控锌指蛋白281表达对黑素瘤细胞增殖、迁移的影响[J].中华皮肤科杂志,2020,53(9):720-724.
[15] WU X,DAI L,ZHANG Z,et al.Overexpression of microRNA-203 can downregulate survivin and function as a potential therapeutic target in papillary thyroid cancer[J].Oncol Lett,2020,19(1):61-68.
[16] YOU A,FU L,LI Y,et al.MicroRNA-203 restrains epithelial-mesenchymal transition,invasion and migration of papillary thyroid cancer by downregulating AKT3[J].Cell Cycle,2020,19(10):1105-1121.
[17] KAN Q,SU Y,YANG H.MicroRNA-335 is downregulated in papillary thyroid cancer and suppresses cancer cell growth,migration and invasion by directly targeting ZEB2[J].Oncol Lett,2017,14(6):7622-7628.

服务与反馈:
文章下载】【发表评论】【查看评论】【加入收藏
提示:您还未登录,请登录!点此登录
您是第 290842 位访问者


copyright ©《东南大学学报(医学版)》编辑部
联系电话:025-87232481 83272483
电子邮件:
bjb@pub.seu.edu.cn

苏ICP备09058364