Department of Thoracic Medicine Oncology, Hunan Cancer Hospital and The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha 410013, PR China.
Department of Orthopedics, Hunan Cancer Hospital and The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha 410013, PR China.
Int J Biol Macromol. 2020 Dec 1;164:1294-1303. doi: 10.1016/j.ijbiomac.2020.07.172. Epub 2020 Jul 19.
Here, we revealed the novel role of long non-coding RNAs (lncRNAs) SOX21 antisense RNA 1 (SOX21-AS1)/TSPAN8/GATA6 in progression of lung adenocarcinoma. SOX21-AS1 expression was quantified in lung adenocarcinoma tissues and cells by RT-qPCR. Then, gain- and loss-of-function experiments were conducted in lung adenocarcinoma cells. Expression of GATA6, TSPAN8 and extracellular signal-regulated kinase (ERK) signaling pathway-related genes was determined in lung adenocarcinoma cells by western blot analysis. The interaction and relationship among SOX21-AS1, GATA6 and TSPAN8 were predicted and verified respectively by RNA pull down, RIP, ChIP, and dual-luciferase reporter assays. Next, lung adenocarcinoma cell proliferation, colony formation, invasion and migration were assessed by 5-ethynyl-2'-deoxyuridine staining, colony formation assay and Transwell assay. Xenograft tumors were established in nude mice and the tumor growth was observed and recorded. SOX21-AS1 was observed to be highly expressed in lung adenocarcinoma tissues. The overexpression of SOX21-AS1, GATA6 or TSPAN8 obviously enhanced cell biological functions in lung adenocarcinoma. Meanwhile, SOX21-AS1 interacted with GATA6 which bound to TSPAN8 promoter and promoted TSPAN8 expression, which further enhanced cell colony formation, proliferation and invasion, and also activated ERK signaling pathway. Silencing of SOX21-AS1 and inhibiting its binding to GATA6 downregulate TSPAN8 and thereby exert anti-oncogenic effects in lung adenocarcinoma.
在这里，我们揭示了长链非编码 RNA（lncRNA）SOX21 反义 RNA 1（SOX21-AS1）/TSPAN8/GATA6 在肺腺癌进展中的新作用。通过 RT-qPCR 定量检测肺腺癌组织和细胞中的 SOX21-AS1 表达。然后，在肺腺癌细胞中进行了增益和缺失功能实验。通过 Western blot 分析测定肺腺癌细胞中 GATA6、TSPAN8 和细胞外信号调节激酶（ERK）信号通路相关基因的表达。通过 RNA 下拉、RIP、ChIP 和双荧光素酶报告基因测定分别预测和验证 SOX21-AS1、GATA6 和 TSPAN8 之间的相互作用和关系。接下来，通过 5-乙炔基-2'-脱氧尿苷染色、集落形成测定和 Transwell 测定评估肺腺癌细胞的增殖、集落形成、侵袭和迁移。在裸鼠中建立肺腺癌细胞移植瘤，观察并记录肿瘤生长情况。观察到 SOX21-AS1 在肺腺癌组织中高表达。SOX21-AS1、GATA6 或 TSPAN8 的过表达明显增强了肺腺癌细胞的生物学功能。同时，SOX21-AS1 与 GATA6 相互作用，GATA6 结合到 TSPAN8 启动子上并促进 TSPAN8 表达，进一步增强细胞集落形成、增殖和侵袭，并激活 ERK 信号通路。沉默 SOX21-AS1 并抑制其与 GATA6 的结合下调 TSPAN8，从而在肺腺癌中发挥抑癌作用。
