Dong Dongfang, Lun Yue, Sun Bo, Sun Haiyuan, Wang Qunying, Yuan Gang, Quan Jingzi
Department of Gastroenterology, No.971 Hospital of People's Liberation Army Navy, Qingdao, China.
Gen Physiol Biophys. 2020 Jan;39(1):1-12. doi: 10.4149/gpb_2019044.
Gastric cancer (GC) is a high mortality disease. We studied the function and mechanism of long non-coding RNA prostate cancer-associated transcript 6 (lncRNA PCAT6) on cell proliferation and epithelial-mesenchymal transition (EMT) in GC cells. CCK-8, flow cytometry and colony formation assay were respectively used to detect the cell viability, apoptosis and colony formation. PCAT6 and miR-15a expression were changed by cell transfection. Moreover, the level of Cyclin D1, p53, Bax, Cleaved caspase-3 and relate-proteins of EMT and cell pathways were investigated by Western blot. Besides, the level of miR-15a and PCAT6 was tested by RT-qPCR. Besides, the target relation between miR-15a and PCAT6 were tested by luciferase assay. PCAT6 was highly expressed in GC cells and tissues. Silencing of PCAT6 restrained the relate-proteins of cell proliferation and EMT. Furthermore, PCAT6 reversely regulated miR-15a and miR-15a inhibitor reversed the efficacy of sh-PCAT6 in cell proliferation and EMT. PCAT6 restrained the relate-proteins of RB/E2F and Wnt/β-catenin pathways and miR-15a reverse this progress. Finally, PCAT6 was a target of miR-15a. Silencing of lncRNA PCAT6 restrained proliferation and EMT of GC cells by targeting miR-15a via RB/E2F and Wnt/β-catenin pathways.
胃癌（GC）是一种高致死率疾病。我们研究了长链非编码RNA前列腺癌相关转录本6（lncRNA PCAT6）对GC细胞增殖和上皮-间质转化（EMT）的作用及机制。分别采用CCK-8法、流式细胞术和集落形成实验检测细胞活力、凋亡和集落形成情况。通过细胞转染改变PCAT6和miR-15a的表达。此外，采用蛋白质免疫印迹法检测细胞周期蛋白D1、p53、Bax、裂解的半胱天冬酶-3以及EMT和细胞通路相关蛋白的水平。此外，通过RT-qPCR检测miR-15a和PCAT6的水平。此外，通过荧光素酶报告基因实验检测miR-15a与PCAT6之间的靶向关系。PCAT6在GC细胞和组织中高表达。沉默PCAT6可抑制细胞增殖和EMT相关蛋白。此外，PCAT6反向调节miR-15a，miR-15a抑制剂可逆转sh-PCAT6对细胞增殖和EMT的作用效果。PCAT6抑制RB/E2F和Wnt/β-连环蛋白通路相关蛋白，miR-15a可逆转这一过程。最后，PCAT6是miR-15a的靶点。沉默lncRNA PCAT6通过RB/E2F和Wnt/β-连环蛋白通路靶向miR-15a，从而抑制GC细胞的增殖和EMT。
