Jiang Hui, Liang Min, Jiang Yanqiong, Zhang Ting, Mo Kexin, Su Suwen, Wang Aiping, Zhu Yongyi, Huang Guanqun, Zhou Rujian
1Department of Abdominal Oncology, The Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510700 Guangdong China.
2Department of Gynaecology, The Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510700 Guangdong China.
Cancer Cell Int. 2019 May 31;19:152. doi: 10.1186/s12935-019-0872-4. eCollection 2019.
Recently, lncRNA-Testis developmental related gene 1 (TDRG1) was proved to be a key modulator in reproductive organ-related cancers. The biological role of TDRG1 in cervical cancer (CC) progression remains largely unknown.
Real-time PCR (qRT-PCR) examined the expression level of TDRG1, microRNA (miR)-326 and MAPK1 mRNA. OS tissues and corresponding relative normal tissues, as well as CC cell lines and normal cell line Ect1/E6E7 were collected to determine the expression of TDRG1 in CC. MTT, colony formation, wound-healing, transwell and flow cytometer assay detected the influence of TDRG1 and miR-326 on CC cells growth, metastasis and apoptosis. Western blot examined proteins level. Bioinformatics, RNA pull-down assay, RNA immunoprecipitation and dual-luciferase reporter assays detected the molecular mechanism of TDRG1 in CC. Xenograft tumour model was established to determine the role of TDRG1 in vivo.
The expression of TDRG1 was significantly increased in CC tissues and cell lines compared with normal tissue and normal cell line respectively and its expression was associated with clinicopathological characteristics of CC patients. Knockdown of TDRG1 inhibited the cell proliferation, migration and invasion in Hela and SIHA cells. Moreover, TDRG1 directly interacted with miR-326, and the inhibition effect on cell growth and metastasis induced by TDRG1 siRNA can be abrogated by miR-326 silencing by its inhibitor in Hela and SIHA cells. Further, MAPK1 was proved to be a direct target of miR-326, and its expression was negatively regulated by miR-326 while positively modulated by TDRG1.
TDRG1 acts as a competing endogenous lncRNA (ceRNA) to modulate MAPK1 by sponging miR-326 in CC, shedding new light on TDRG1-directed diagnostics and therapeutics in CC.
最近，长链非编码RNA-睾丸发育相关基因1（TDRG1）被证明是生殖器官相关癌症的关键调节因子。TDRG1在宫颈癌（CC）进展中的生物学作用仍 largely未知。
实时荧光定量PCR（qRT-PCR）检测TDRG1、微小RNA（miR）-326和丝裂原活化蛋白激酶1（MAPK1）mRNA的表达水平。收集OS组织及其相应的相对正常组织，以及CC细胞系和正常细胞系Ect1/E6E7，以确定TDRG1在CC中的表达。MTT法、集落形成实验、伤口愈合实验、Transwell实验和流式细胞仪检测TDRG1和miR-326对CC细胞生长、转移和凋亡的影响。蛋白质印迹法检测蛋白质水平。生物信息学、RNA下拉实验、RNA免疫沉淀实验和双荧光素酶报告基因实验检测TDRG1在CC中的分子机制。建立异种移植肿瘤模型以确定TDRG1在体内的作用。
与正常组织和正常细胞系相比，TDRG1在CC组织和细胞系中的表达分别显著增加，且其表达与CC患者的临床病理特征相关。敲低TDRG1可抑制Hela和SIHA细胞的增殖、迁移和侵袭。此外，TDRG1直接与miR-326相互作用，在Hela和SIHA细胞中，其抑制剂沉默miR-326可消除TDRG1 siRNA对细胞生长和转移的抑制作用。进一步研究表明，MAPK1是miR-326的直接靶点，其表达受miR-326负调控，而受TDRG1正调控。
在CC中，TDRG1作为一种竞争性内源性长链非编码RNA（ceRNA），通过海绵吸附miR-326来调节MAPK1，为CC中TDRG1指导的诊断和治疗提供了新的思路。
