Shi Qizhong, Wang Yongling, Mu Yonghui, Wang Xin, Fan Qingxia
Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Department of cardiothoracic Surgery, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang, China.
Cell Physiol Biochem. 2018;46(5):2187-2196. doi: 10.1159/000489548. Epub 2018 Apr 28.
BACKGROUND/AIMS: MicroRNAs (miRNAs) are non-coding single stranded RNAs of 17-25 nucleotides in size, and their altered expression has been observed in various cancers. Previous studies have confirmed that miR-433-3p has effects on cancer cell proliferation, invasion, and migration, and its expression also correlates with sensitivity to chemotherapy. However, to date, there have been no studies on the biological functions of miR-433-3p in esophageal squamous cell carcinoma (ESCC).
The Cell Counting Kit-8, transwell, and matrigel assays were used to test the effects of miR-433-3p and its predicted target, growth factor receptor-bound protein 2 (GRB2), on the proliferation, migration, and invasion of Eca109 and KYSE30 cells, two types of esophageal cancer cell lines. The miR-433-3p binding site in the 3' untranslated region (UTR) region of GRB2 was predicted and verified using miRNA target site prediction software and structuring correct mutant examination. Western blotting and fluorescent quantitative PCR (FQ-PCR) techniques were employed to evaluate GRB2 expression. The inhibitory effects of miR-433-3p on tumor growth were investigated using a tumor xenograft model.
The binding site of miR-433-3p was identified in the 3'UTR region of GRB2. Western blotting and FQ-PCR showed that miR-433-3p inhibited the mRNA and protein expression of GRB2. Overexpression of GRB2 inhibited tumorigenesis in nude mice. MiR-433-3p overexpression inhibited the proliferation, migration, and invasion of ESCC cells by suppressing GRB2 gene expression.
Our findings suggest that targeting miR-433-3p may have therapeutic benefits in ESCC.
背景/目的：微小RNA（miRNA）是大小为17 - 25个核苷酸的非编码单链RNA，其表达改变在多种癌症中均有观察到。既往研究证实miR - 433 - 3p对癌细胞的增殖、侵袭和迁移有影响，其表达还与化疗敏感性相关。然而，迄今为止，尚无关于miR - 433 - 3p在食管鳞状细胞癌（ESCC）中的生物学功能的研究。
使用细胞计数试剂盒 - 8、Transwell和基质胶实验检测miR - 433 - 3p及其预测靶点生长因子受体结合蛋白2（GRB2）对两种食管癌细胞系Eca109和KYSE30细胞增殖、迁移和侵袭的影响。使用miRNA靶点预测软件并构建正确的突变体进行检测，预测并验证GRB2的3'非翻译区（UTR）区域中的miR - 433 - 3p结合位点。采用蛋白质免疫印迹法和荧光定量PCR（FQ - PCR）技术评估GRB2的表达。使用肿瘤异种移植模型研究miR - 433 - 3p对肿瘤生长的抑制作用。
在GRB2的3'UTR区域鉴定出miR - 433 - 3p的结合位点。蛋白质免疫印迹法和FQ - PCR显示miR - 433 - 3p抑制GRB2的mRNA和蛋白表达。GRB2的过表达抑制裸鼠肿瘤发生。miR - 433 - 3p的过表达通过抑制GRB2基因表达抑制ESCC细胞的增殖、迁移和侵袭。
我们的研究结果表明，靶向miR - 433 - 3p可能对ESCC有治疗益处。
