Zheng Xiao-Ying, Cao Ming-Zheng, Ba Ying, Li Yue-Feng, Ye Jun-Ling
Department of Pathology, Qinghai University Affiliated Hospital, Xining, Qinghai, China.
Department of General Surgery, Linyi Central Hospital, Linyi, Shandong, China.
Cancer Biomark. 2021;31(1):1-11. doi: 10.3233/CBM-201709.
Long non-coding RNA testis-specific transcript, Y-linked 15 (TTTY15) is oncogenic in prostate cancer, however its expression and function in colorectal cancer remain largely unknown.
Paired colorectal cancer samples/normal tissues were collected, and the expression levels of TTTY15, miR-29a-3p and disheveled segment polarity protein 3 (DVL3) were examined by quantitative real-time polymerase chain reaction (qRT-PCR); TTTY15 shRNA and overexpression plasmids were transfected into HT29 and HCT-116 cell lines using lipofectamine reagent, respectively; the proliferation and colony formation were detected by CCK-8 assay and plate colony formation assay; qRT-PCR and Western blot were used to analyze the changes of miR-29a-3p and DVL3; dual-luciferase reporter gene assay was used to determine the regulatory relationships between miR-29a-3p and TTTY15, miR-29a-3p and DVL3.
TTTY15 was significantly up-regulated in cancerous tissues of colorectal cancer samples, positively correlated with the expression of DVL3, while negatively correlated with the expression of miR-29a-3p. After TTTY15 shRNAs were transfected into colorectal cancer cells, the proliferation and metastasis of cancer cells were significantly inhibited, while TTTY15 overexpression had opposite biological effects. TTTY15 shRNA could reduce the expression of DVL3 on both mRNA and protein levels, and the luciferase activity of TTTY15 sequence was also inhibited by miR-29a-3p. DVL3 was also validated as a target gene of miR-29a-3p, and it could be repressed by miR-29a-3p mimics or TTTY15 shRNA.
TTTY15 is abnormally upregulated in colorectal cancer tissues, and it can modulate the proliferation and metastasis of colorectal cancer cells. It functions as the ceRNA to regulate the expression of DVL3 by sponging miR-29a-3p.
长链非编码RNA睾丸特异性转录本,Y连锁15(TTTY15)在前列腺癌中具有致癌性,然而其在结直肠癌中的表达及功能仍 largely未知。
收集配对的结直肠癌样本/正常组织,采用定量实时聚合酶链反应(qRT-PCR)检测TTTY15、miR-29a-3p和无序段极性蛋白3(DVL3)的表达水平;分别使用脂质体试剂将TTTY15 shRNA和过表达质粒转染至HT29和HCT-116细胞系;采用CCK-8法和平板集落形成试验检测细胞增殖和集落形成;运用qRT-PCR和蛋白质印迹法分析miR-29a-3p和DVL3的变化;采用双荧光素酶报告基因试验确定miR-29a-3p与TTTY15、miR-29a-3p与DVL3之间的调控关系。
TTTY15在结直肠癌样本的癌组织中显著上调,与DVL3的表达呈正相关,而与miR-29a-3p的表达呈负相关。将TTTY15 shRNAs转染至结直肠癌细胞后,癌细胞的增殖和转移受到显著抑制,而TTTY15过表达则具有相反的生物学效应。TTTY15 shRNA可在mRNA和蛋白质水平降低DVL3的表达,miR-29a-3p也可抑制TTTY15序列的荧光素酶活性。DVL3也被验证为miR-29a-3p的靶基因,miR-29a-3p模拟物或TTTY15 shRNA均可抑制其表达。
TTTY15在结直肠癌组织中异常上调,可调节结直肠癌细胞的增殖和转移。它作为竞争性内源RNA,通过海绵吸附miR-29a-3p来调节DVL3的表达。