Abstract：
OBJECTIVE: The purpose of this study was to explore the role of RAE1 in the invasion and metastasis of gastric cancer (GC) cells.
METHODS: RAE1 expression in GC cells was determined by reverse-transcription polymerase chain reaction (qRT-PCR) and Western blotting (WB). Cell models featuring RAE1 gene silencing and overexpression were constructed by lentiviral transfection; The proliferation, migration, and invasion ability of cells were detected by cell counting, colony formation assay, would healing assay, and transwell invasion and migration test. WB analysis of ERK/MAPK signaling pathway (ERK1/2, p-ERK1/2, c-Myc) and EMT-related molecules (ZEB1, E-cadherin, N-cadherin, and Vimentin).
RESULTS: The expression level of RAE1 in GC was notably higher than in adjacent tissues. Elevated RAE1 expression correlated with an unfavorable prognosis for GC patients. Knockdown of RAE1, as compared to the control group, resulted in a significant inhibition of proliferation, migration, and invasion abilities in GC cell lines. Furthermore, RAE1 knockdown led to a substantial decrease in the expression of N-cadherin, vimentin, ZEB1, p-ERK1/2, and c-Myc proteins, coupled with a marked increase in E-cadherin expression. The biological effects of RAE1 in GC cells were effectively reversed by the inhibition of the ERK/MAPK signaling pathway using SCH772984. Additionally, RAE1 knockdown demonstrated a suppressive effect on GC tumor size in vivo. Immunohistochemistry (IHC) results revealed significantly lower expression of Ki-67 in RAE1 knockout mice compared to the control group.
CONCLUSIONS: RAE1 promotes GC cell migration and invasion through the ERK/MAPK pathway and is a potential therapeutic target for GC therapy.
METHODS: RAE1 expression in GC cells was determined by reverse-transcription polymerase chain reaction (qRT-PCR) and Western blotting (WB). Cell models featuring RAE1 gene silencing and overexpression were constructed by lentiviral transfection; The proliferation, migration, and invasion ability of cells were detected by cell counting, colony formation assay, would healing assay, and transwell invasion and migration test. WB analysis of ERK/MAPK signaling pathway (ERK1/2, p-ERK1/2, c-Myc) and EMT-related molecules (ZEB1, E-cadherin, N-cadherin, and Vimentin).
RESULTS: The expression level of RAE1 in GC was notably higher than in adjacent tissues. Elevated RAE1 expression correlated with an unfavorable prognosis for GC patients. Knockdown of RAE1, as compared to the control group, resulted in a significant inhibition of proliferation, migration, and invasion abilities in GC cell lines. Furthermore, RAE1 knockdown led to a substantial decrease in the expression of N-cadherin, vimentin, ZEB1, p-ERK1/2, and c-Myc proteins, coupled with a marked increase in E-cadherin expression. The biological effects of RAE1 in GC cells were effectively reversed by the inhibition of the ERK/MAPK signaling pathway using SCH772984. Additionally, RAE1 knockdown demonstrated a suppressive effect on GC tumor size in vivo. Immunohistochemistry (IHC) results revealed significantly lower expression of Ki-67 in RAE1 knockout mice compared to the control group.
CONCLUSIONS: RAE1 promotes GC cell migration and invasion through the ERK/MAPK pathway and is a potential therapeutic target for GC therapy.
摘要：
目的：本研究的目的是探讨RAE1在胃癌（GC）细胞侵袭和转移中的作用。
方法：通过逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹（WB）检测GC细胞中RAE1的表达。通过慢病毒转染构建RAE1基因沉默和过表达的细胞模型;迁移,细胞计数检测细胞的侵袭能力，集落形成试验,会愈合试验，以及Transwell入侵和迁移测试。WB分析ERK/MAPK信号通路（ERK1/2，p-ERK1/2，c-Myc）和EMT相关分子（ZEB1，E-cadherin，N-钙黏着蛋白,和Vimentin）。
结果：RAE1在GC中的表达水平明显高于癌旁组织。RAE1表达升高与GC患者的不良预后相关。与对照组相比，RAE1的击倒，导致增殖的显著抑制，迁移,和GC细胞系的侵袭能力。此外，RAE1敲低导致N-cadherin的表达大幅下降，波形蛋白,ZEB1、p-ERK1/2和c-Myc蛋白，再加上E-cadherin表达的显著增加。通过使用SCH772984抑制ERK/MAPK信号通路，可以有效逆转RAE1在GC细胞中的生物学效应。此外,RAE1敲低显示了对体内GC肿瘤大小的抑制作用。免疫组织化学（IHC）结果显示，与对照组相比，RAE1敲除小鼠中Ki-67的表达显着降低。
结论：RAE1通过ERK/MAPK通路促进GC细胞迁移和侵袭，是GC治疗的潜在治疗靶点。
方法：通过逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹（WB）检测GC细胞中RAE1的表达。通过慢病毒转染构建RAE1基因沉默和过表达的细胞模型;迁移,细胞计数检测细胞的侵袭能力，集落形成试验,会愈合试验，以及Transwell入侵和迁移测试。WB分析ERK/MAPK信号通路（ERK1/2，p-ERK1/2，c-Myc）和EMT相关分子（ZEB1，E-cadherin，N-钙黏着蛋白,和Vimentin）。
结果：RAE1在GC中的表达水平明显高于癌旁组织。RAE1表达升高与GC患者的不良预后相关。与对照组相比，RAE1的击倒，导致增殖的显著抑制，迁移,和GC细胞系的侵袭能力。此外，RAE1敲低导致N-cadherin的表达大幅下降，波形蛋白,ZEB1、p-ERK1/2和c-Myc蛋白，再加上E-cadherin表达的显著增加。通过使用SCH772984抑制ERK/MAPK信号通路，可以有效逆转RAE1在GC细胞中的生物学效应。此外,RAE1敲低显示了对体内GC肿瘤大小的抑制作用。免疫组织化学（IHC）结果显示，与对照组相比，RAE1敲除小鼠中Ki-67的表达显着降低。
结论：RAE1通过ERK/MAPK通路促进GC细胞迁移和侵袭，是GC治疗的潜在治疗靶点。
