Wei Weiyuan, Cao Wenlong, Zhan Zexu, Yan Linhai, Xie Yubo, Xiao Qiang
Department of Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning, People's Republic of China.
Department of Gastrointestinal Surgery, Affiliated Tumor Hospital of Guangxi Medical University, Nanning, People's Republic of China.
Onco Targets Ther. 2019 May 21;12:3965-3976. doi: 10.2147/OTT.S191015. eCollection 2019.
MicroRNAs (miRNAs) play a key role in the development of gastric cancer (GC). MiRNA arrays showed that lymph node metastasis in GC is correlated with the expression of miR-1284. Although its function and mechanisms in GC have not been fully described, the regulation of EIF4A1 by miR-1284 and its role in drug-resistant GC has been reported in our previous studies. qRT-PCR was used to study the level of miR-1284 expression in GC cell lines and tissues. Subsequently, the CCK-8 assay was used to detect cell proliferation, while transwell assay was used to detect invasion and migration of the GC cells. Flow cytometry was used to detect the effect of miR-1284 on GC cells in vivo by building subcutaneous GC nude mice transplantation tumor model. In addition, the influence of miR-1284 gene expression profile in SGC-7901 cells was detected by total gene expression chip, and the target gene of miR-1284 was detected by luciferase reporter assay, qRT-PCR, and western blotting. The miR-1284 level was down-regulated in GC tssues and cell lines. MiR-1284 was significantly associated with tumor size, degree of differentiation and patients' distant metastasis. MiR-1284 inhibited invasion, migration, and proliferation of GC cells. During the G1/S phase, miR-1284 arrested the cycle of GC cells in vitro. MiR-1284 also suppressed tumor from growing and metastasizing in xenograft models as well as influenced the gene expression profile in SGC-7901 cells. Also, EIF4A1 was the direct target gene for miR-1284. Further, an inverse correlation between the miR-1284 expression and EIF4A1 was found in GC tissues. Over-expressed miR-1284 decreased c-Myc, MMP12, JUN expression, while increased CDH1 expression. These data suggested that miR-1284 acts as a tumor suppressor, and directly blocked EIF4A1 in GC.
微小RNA（miRNA）在胃癌（GC）的发展中起关键作用。miRNA阵列显示，GC中的淋巴结转移与miR-1284的表达相关。尽管其在GC中的功能和机制尚未完全阐明，但我们先前的研究报道了miR-1284对真核翻译起始因子4A1（EIF4A1）的调控及其在耐药GC中的作用。采用qRT-PCR研究miR-1284在GC细胞系和组织中的表达水平。随后，使用CCK-8法检测细胞增殖，而transwell法用于检测GC细胞的侵袭和迁移。通过构建皮下GC裸鼠移植瘤模型，利用流式细胞术检测miR-1284对体内GC细胞的影响。此外，通过全基因表达芯片检测miR-1284基因表达谱在SGC-7901细胞中的影响，并通过荧光素酶报告基因检测、qRT-PCR和蛋白质免疫印迹法检测miR-1284的靶基因。miR-1284水平在GC组织和细胞系中下调。miR-1284与肿瘤大小、分化程度及患者远处转移显著相关。miR-1284抑制GC细胞的侵袭、迁移和增殖。在G1/S期，miR-1284在体外使GC细胞周期停滞。miR-1284在异种移植模型中也抑制肿瘤生长和转移，并影响SGC-7901细胞中的基因表达谱。此外，EIF4A1是miR-1284的直接靶基因。进一步研究发现，GC组织中miR-1284表达与EIF4A1呈负相关。过表达miR-1284可降低c-Myc、基质金属蛋白酶12（MMP12）、原癌基因蛋白（JUN）的表达，同时增加钙黏蛋白1（CDH1）的表达。这些数据表明，miR-1284作为一种肿瘤抑制因子，在GC中直接阻断EIF4A1。
