Department of Urology, Yiwu Central Hospital, Yiwu, Zhejiang, China.
Eur Rev Med Pharmacol Sci. 2019 Sep;23(18):7816-7825. doi: 10.26355/eurrev_201909_18991.
Prostatic cancer (PCa) is a common cancer in males. Long non-coding RNA (lncRNA) TTN-AS1 has been reported as an oncogene in diverse cancers. This study aimed to explore the functions and mechanism of TTN-AS1 in PCa.
The levels of TTN-AS1 and miR-193a-5p in PCa cells (DU145, PC3, 22RV1, C4-2B, and LNCaP) were measured by qRT-PCR. The putative target of TTN-AS1 was predicted by starBase v2.0 online database, and this interaction was validated by Dual-Luciferase reporter assay. The cell viability and apoptosis rate in DU145 and PC3 cells were assessed by MTT assay and flow cytometry, respectively. The protein levels of CyclinD1, p21, p27, Bcl-2, Bax, and cleaved-caspase3 were detected by Western blot.
The relative expression of TTN-AS1 was apparently up-regulated, and the level of miR-193a-5p was strikingly down-regulated in PCa cells. The interaction between TTN-AS1 and miR-193a-5p was predicted by starBase v2.0 online database and verified by Dual-Luciferase reporter assay. The functional experiments indicated that TTN-ASI knockdown or miR-193a-5p inhibited cell viability and induced cell apoptosis rate in DU145 and PC3 cells. Furthermore, the recuperated experiments exhibited that miR-193a-5p inhibitor counteracted the inhibitory effect on cell viability and the promotion effect on cell apoptosis rate in DU145 and PC3 cells induced by TTN-AS1 silencing.
These data indicated that TTN-AS1 was dramatically up-regulated, and miR-193a-5p was significantly down-regulated in PCa cells. The functional and mechanistical experiments unraveled that lncRNA TTN-AS1 sponged miR-193a-5p to promote cell proliferation and repress cell apoptosis in prostatic cancer, and this new regulatory pathway may shed light on the mechanism of prostatic cancer.
前列腺癌（PCa）是男性常见的癌症。长链非编码 RNA（lncRNA）TTN-AS1 已被报道为多种癌症的癌基因。本研究旨在探讨 TTN-AS1 在 PCa 中的功能和机制。
通过 qRT-PCR 测量 PCa 细胞（DU145、PC3、22RV1、C4-2B 和 LNCaP）中 TTN-AS1 和 miR-193a-5p 的水平。通过 starBase v2.0 在线数据库预测 TTN-AS1 的可能靶标，并通过双荧光素酶报告基因实验验证该相互作用。通过 MTT 测定和流式细胞术分别评估 DU145 和 PC3 细胞的细胞活力和细胞凋亡率。通过 Western blot 检测细胞周期蛋白 D1、p21、p27、Bcl-2、Bax 和 cleaved-caspase3 的蛋白水平。
PCa 细胞中 TTN-AS1 的相对表达明显上调，miR-193a-5p 的水平明显下调。starBase v2.0 在线数据库预测 TTN-AS1 与 miR-193a-5p 之间存在相互作用，并通过双荧光素酶报告基因实验验证。功能实验表明，TTN-ASI 敲低或 miR-193a-5p 抑制 DU145 和 PC3 细胞的细胞活力并诱导细胞凋亡率。此外，恢复实验表明，miR-193a-5p 抑制剂逆转了 TTN-AS1 沉默对 DU145 和 PC3 细胞活力的抑制作用和对细胞凋亡率的促进作用。
这些数据表明，PCa 细胞中 TTN-AS1 明显上调，miR-193a-5p 明显下调。功能和机制实验表明，lncRNA TTN-AS1 通过海绵吸附 miR-193a-5p 促进前列腺癌细胞增殖并抑制细胞凋亡，这一新的调控途径可能为前列腺癌的机制提供新的思路。
