GADD45G Interacts with E-cadherin to Suppress the Migration and Invasion of Esophageal Squamous Cell Carcinoma
GADD45G Interacts with E-cadherin to Suppress the Migration and Invasion of Esophageal Squamous Cell Carcinoma
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DOI：
10.1007/s10620-019-05836-8
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发表时间：
2020-04-01
影响因子：
3.1
通讯作者：
Liu, Xuefeng
中科院分区：
文献类型：
--
作者：
Li, Tongtong;Xu, Lele;Liu, Xuefeng
背景/目的食管鳞状细胞癌（ESCC）是最常见的恶性肿瘤之一，预后较差。转移是癌症相关死亡的主要原因。据报道，生长阻滞和DNA损伤诱导的45 γ (GADD45G)与ESCC的存活、侵袭和转移有关。本研究旨在探讨GADD45G在ESCC细胞迁移和侵袭中的作用及其机制。方法通过Transwell法测定GADD45G对ESCC细胞迁移和侵袭的影响及其对E-cadherin功能的需求。GST-pull - down和IP检测GADD45G与E-cadherin的相互作用。RT-qPCR和western blot检测GADD45G过表达后E-cadherin的表达情况。亚细胞分离后，用western blot检测细胞质、细胞核和细胞膜中e -钙粘蛋白的水平。结果低表达GADD45G可增强KYSE150细胞的迁移和侵袭能力，而过表达GADD45G则对YES2和KYSE30细胞的迁移和侵袭能力有相反的影响。GADD45G可与E-cadherin相互作用，提高E-cadherin的膜水平。敲低E-cadherin可消除GADD45G对ESCC细胞迁移和侵袭的抑制作用。有趣的是，二聚体解离突变体GADD45G不能与E-cadherin相互作用，几乎失去了抑制ESCC细胞迁移和侵袭的能力。结论本研究揭示了GADD45G在抑制ESCC细胞迁移和侵袭中的新作用，为了解ESCC转移机制提供了新的思路。
Background/Aims Esophageal squamous cell carcinoma (ESCC) is one of the most prevalent cancers with poor prognosis. Metastasis is the leading cause of cancer-related deaths. The growth arrest and DNA damage-inducible 45 gamma (GADD45G) has been reported to correlate with survival, invasion, and metastasis of ESCC. This study was aimed to investigate the role and mechanism of GADD45G in ESCC cell migration and invasion. Methods Both the effects of GADD45G and its need for E-cadherin to function on ESCC cell migration and invasion were determined through loss- and gain-of-function approaches via Transwell assays. The interaction between GADD45G and E-cadherin was detected by GST-pull down and IP assays. The expression of E-cadherin upon GADD45G overexpression was evaluated by RT-qPCR and western blot. The level of E-cadherin in cytoplasmic, nuclear, and membrane fractions was examined by western blot following subcellular fractionation. Results Knockdown of GADD45G increased the migration and invasion abilities of KYSE150 cells, while overexpression of GADD45G showed the opposite effects on YES2 and KYSE30 cells. GADD45G could interact with E-cadherin and enhanced its membrane level. Knockdown of E-cadherin abolished the inhibitory effects of GADD45G on ESCC cell migration and invasion. Intriguingly, dimer-dissociating mutant of GADD45G could not interact with E-cadherin and almost lost its ability to suppress the ESCC cell migration and invasion. Conclusions This study reveals a novel role for GADD45G in inhibiting the ESCC cell migration and invasion, which will provide a new insight in understanding the ESCC metastatic mechanism.