本研究旨在探讨长链非编码RNA睾丸特异性转录本的调控作用及相关机制。Y连锁15(TTTY15)在胃癌(GC)细胞增殖中的作用,迁移,入侵,细胞凋亡和上皮间质转化(EMT)。通过实时定量PCR(qRT-PCR)检测GC组织和细胞中TTTY15的表达,并分析TTTY15表达与GC临床病理指标的相关性。细胞计数试剂盒-8(CCK-8),BrdU,流式细胞术和Transwell分析用于检测GC细胞增殖,迁移,侵袭和凋亡。Westernblot检测EMT相关蛋白(N-cadherin和E-cadherin)的表达,Wnt家族成员1(Wnt1)蛋白和β-catenin蛋白。生物信息学分析进行预测,并进行了RNA免疫沉淀(RIP)测定和双荧光素酶报告基因测定,以验证microRNAlet-7a-5p(let-7a-5p)与TTTY15和Wnt1mRNA3UTR的靶向关系。发现TTTY15在GC组织和细胞中表达显著上调,并与晚期TNM分期和肿瘤分化差有关。TTTY15过表达促进GC细胞增殖,移民和入侵,N-钙黏着蛋白的表达,Wnt1和β-连环蛋白,并抑制细胞凋亡和E-cadherin的表达,而击倒TTTY15则产生相反的效果。TTTY15直接靶向let-7a-5p并负调控其表达。Wnt1是let-7a-5p的靶基因,TTTY15可以间接和正向调节Wnt1的表达。总之,TTTY15促进GC进展,通过调节let-7a-5p/Wnt1轴来激活Wnt/β-catenin通路。
The present study is aimed to investigate the regulatory effects and related mechanism of long non-coding RNA testis-specific transcript, Y-linked 15 (
TTTY15) in gastric carcinoma (GC) cell proliferation, migration, invasion, apoptosis and epithelial-mesenchymal transition (EMT).
TTTY15 expression in GC tissue samples and cells was detected by quantitative real-time PCR (qRT-PCR), and the correlation between
TTTY15 expression and GC clinicopathological indicators was analyzed. Cell counting kit-8 (CCK-8), BrdU, flow cytometry and Transwell assays were performed for detecting GC cell proliferation, migration, invasion and apoptosis. Western blot was performed for detecting the expressions of EMT-associated proteins (N-cadherin and E-cadherin), Wnt family member 1 (Wnt1) protein and β-catenin protein. Bioinformatics analysis was conducted to predict, and RNA immunoprecipitation (RIP) assay and dual-luciferase reporter gene assay were performed to verify the targeted relationships of microRNA let-7a-5p (let-7a-5p) with TTTY15 and Wnt1 mRNA 3\'UTR. It was found that TTTY15 expression was significantly up-regulated in GC tissues and cells, and was associated with advanced TNM stage and poor tumor differentiation. TTTY15 overexpression promoted GC cell proliferation, migration and invasion, the expressions of N-cadherin, Wnt1 and β-catenin protein, and inhibited the apoptosis and E-cadherin expression, while knocking down TTTY15 had the opposite effects.
TTTY15 directly targeted let-7a-5p and negatively regulated its expression. Wnt1 was the target gene of let-7a-5p, and TTTY15 could indirectly and positively regulate Wnt1 expression. In conclusion,
TTTY15 promotes GC progression, by regulating the let-7a-5p/Wnt1 axis to activate the Wnt/β-catenin pathway.