TIPE3调控AKT/ERK-GSK3β-β-catenin/Snail轴促肺癌生长、转移的作用

Recently, a seminal study has shown that tumor necrosis factor (TNF)-alpha-induced protein 8-like 3 (TNFAIP8L3, TIPE3), as a newly identified transfer protein of phosphoinositide second messengers such as phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) and phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3), has a crucial role for promoting AKT and ERK signaling. The effect of TIPE3 on non-small cell lung cancer (NSCLC) is still poorly understood so far. Our preliminary studies have shown that TIPE3 remarkably promotes the growth and the metastasis of human NSCLC. However, its underlying mechansm is largly unclear. We thus hypothesized that TIPE3 would modulate the AKT/ERK-GSK3β-β-catenin/Snail axis and then increased the stability and the nuclear translocation of β-catenin and Snail1/2 very possibly by upregulating AKT and ERK signaling, leading to the NSCLC progression. To validate the hypothesis, we established the TIPE3-overexpressing or the TIPE3-shRNA knockdown NSCLC transgenic cells by lentivirus-mediated transfection. We then investigated the effect of TIPE3 overexpression or knockdown on the growth and the metastasis of human NSCLC in in vitro cell model and in in vivo BALB/c nude mouse model using the assays of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), flow cytometry (FCM), wounding healing, Transwell chamber invasion, in vivo bioluminescence image etc, respectively. We also elucidated the underlying mechanism, whether TIPE3-mediated regulation of AKT/ERK-GSK3β-β-catenin/Snail axis is a vital mechanism for NSCLC progression, by the assays of Western blot, kinase/proteasome inhibition, E3 ubiquitin ligase siRNA. Furthermore, we explored the clinical correlation of TIPE3 expression and pathological factors of NSCLC patients or expression of AKT/ERK-GSK3β-β-catenin/Snail axis-related proteins. This project will provide the new ideas for targeting TIPE3 in the prvention and therapy of NSCLC.

研究显示TIPE3是一种磷酸肌醇转运蛋白，在促进AKT和ERK信号中起关键作用。迄今TIPE3对非小细胞肺癌（NSCLC）的作用知之甚少。我们预实验发现TIPE3促进NSCLC生长、转移，但机制尚不明确。为此我们提出假说：TIPE3可能通过上调AKT、ERK信号调控AKT/ERK-GSK3β-β-catenin/Snail轴影响β-catenin、Snail1/2，参与NSCLC进展。为验证该假说，本项目构建慢病毒介导的TIPE3过表达、沉默NSCLC细胞，采用MTT、FCM、划痕、侵袭、Western blot、免疫组化、激酶/蛋白酶体抑制、siRNA等方法，在细胞、动物、临床标本水平探讨TIPE3对NSCLC的作用及其与病理因素的关系，明确TIPE3调控AKT/ERK-GSK3β-β-catenin/Snail轴促进NSCLC的作用机制，为以TIPE3为靶标防治NSCLC提供新的思路。