Department of Clinical Laboratory, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, PR China.
Department of Emergency, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, PR China.
Life Sci. 2020 May 15;249:117478. doi: 10.1016/j.lfs.2020.117478. Epub 2020 Feb 28.
Gastric cancer is a severe disease with a high occurrence rate worldwide. And lncRNAs are demonstrated to be responsible for cancer growth and metastasis. So, it is of great importance to explore the lncRNAs involved mechanism of gastric cancer occurrence and development deeply.
Transfection was conducted to build over-expression and down-expression models. Moreover, RT-qPCR and western blot were used to detect the transcriptional and translational levels. The biological functions such as proliferation, migration and invasion of AGS cells were evaluated by MTT analysis, colony formation assay, scarification detection and transwell assay, respectively. The potential binding of miR-135b and its downstream and upstream molecules was validated by dual luciferase reporter gene assay or RIP. Also, the in-vivo mice model was further used to demonstrate the role of lncRNA PCAT18 in gastric cancer.
PCAT18 down-expression promoted proliferation, migration and invasion of gastric cancer cells. Furtherly, over-expression of miR-135b also promoted these biological characteristics of AGS cells. Importantly, we found that PCAT18 could bind miR-135b which also was bound with CLDN11. We found that miR-135b is negatively correlated with CLDN11; PCAT18 and CLDN11 are positively correlated. Moreover, miR-135b mimics could down-regulate protein level of CLDN11, whereas CLDN11 could reverse this effect. In in-vivo experiment, PCAT18 over-expression restrained tumor growth and metastasis.
Over-expressed lncRNA PCAT18 inhibits proliferation, migration and invasion of gastric cancer cells through regulation of miR-135b/CLDN11.
胃癌是一种全球发病率较高的严重疾病。lncRNAs 被证明与癌症的生长和转移有关。因此，深入探讨 lncRNAs 在胃癌发生和发展中的作用机制具有重要意义。
通过转染构建过表达和下调模型。此外，采用 RT-qPCR 和 Western blot 检测转录和翻译水平。通过 MTT 分析、集落形成实验、划痕检测和 Transwell 实验分别评估 AGS 细胞的增殖、迁移和侵袭等生物学功能。通过双荧光素酶报告基因实验或 RIP 验证 miR-135b 及其下游和上游分子的潜在结合。此外，进一步使用体内小鼠模型来证明 lncRNA PCAT18 在胃癌中的作用。
PCAT18 的下调促进了胃癌细胞的增殖、迁移和侵袭。此外，miR-135b 的过表达也促进了 AGS 细胞的这些生物学特性。重要的是，我们发现 PCAT18 可以与 miR-135b 结合，而 miR-135b 也与 CLDN11 结合。我们发现 miR-135b 与 CLDN11 呈负相关；PCAT18 与 CLDN11 呈正相关。此外，miR-135b 模拟物可以下调 CLDN11 的蛋白水平，而 CLDN11 可以逆转这种效应。在体内实验中，PCAT18 的过表达抑制了肿瘤的生长和转移。
过表达的 lncRNA PCAT18 通过调节 miR-135b/CLDN11 抑制胃癌细胞的增殖、迁移和侵袭。
