Liu Ying, Tang Yanping, Li Ping
Second Department of Gastroenterology, Tianjin Integrated Traditional Chinese and Western Medicine Hospital, Tianjin Nankai Hospital, Tianjin 300100, P.R. China.
College of Acupuncture and Massage, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, P.R. China.
Exp Ther Med. 2018 Dec;16(6):4959-4966. doi: 10.3892/etm.2018.6820. Epub 2018 Oct 2.
The aim of the present study was to examine microRNA (miRNA or miR)-455-5p expression in esophageal squamous cell carcinoma (ESCC) at the tissue and cellular levels in order to elucidate its biological roles. A total of 60 patients with ESCC were enrolled in the present study and reverse transcription-quantitative polymerase chain reaction was used to measure the expression of miR-455-5p. ESCC Eca109 cells were transfected with miR-NC, miR-455-5p mimics or inhibitor and a Cell Counting Kit-8 assay was used to assess proliferation. To investigate the migration and invasion abilities of Eca109 cells, Transwell and Matrigel assays were performed. Western blotting was employed to measure Rab31 protein expression, while a rescue assay was utilized to study the biological roles of miR-455-5p and Rab31 in Eca109 cells. To determine whether Rab31 is a direct target of miR-455-5p, a dual luciferase reporter assay was performed. The results revealed that miR-455-5p expression was decreased in ESCC tissues and was negatively correlated with metastasis and pathogenesis. overexpression of miR-455-5p inhibited the proliferation, migration and invasion of ESCC Eca109 cells. Furthermore, miR-455-5p regulated the expression of Rab31 protein in Eca109 cells. Rab31 overexpression promoted the proliferation, migration and invasion of Eca109 cells. Luciferase reporter assay results revealed that miR-455-5p is able to bind with the 3'-untranslated region of Rab31 mRNA to regulate its expression. In summary, the results of the present study suggest that miR-455-5p expression is decreased in ESCC tissues and is miR-455-5p is negatively correlated with lymphatic metastasis and differentiation. As a tumor-suppressor gene, miR-455-5p inhibits the proliferation, migration and invasion of ESCC Eca109 cells by suppressing the expression of Rab31.
本研究的目的是在组织和细胞水平上检测食管鳞状细胞癌（ESCC）中微小RNA（miRNA或miR）-455-5p的表达，以阐明其生物学作用。本研究共纳入60例ESCC患者，采用逆转录-定量聚合酶链反应检测miR-455-5p的表达。用miR-NC、miR-455-5p模拟物或抑制剂转染ESCC Eca109细胞，并用细胞计数试剂盒-8法评估细胞增殖。为了研究Eca109细胞的迁移和侵袭能力，进行了Transwell和基质胶实验。采用蛋白质免疫印迹法检测Rab31蛋白表达，同时利用拯救实验研究miR-455-5p和Rab31在Eca109细胞中的生物学作用。为了确定Rab31是否为miR-455-5p的直接靶点，进行了双荧光素酶报告基因实验。结果显示，miR-455-5p在ESCC组织中的表达降低，且与转移和发病机制呈负相关。miR-455-5p的过表达抑制了ESCC Eca109细胞的增殖、迁移和侵袭。此外，miR-455-5p调节Eca109细胞中Rab31蛋白的表达。Rab31的过表达促进了Eca109细胞的增殖、迁移和侵袭。荧光素酶报告基因实验结果显示，miR-455-5p能够与Rab31 mRNA的3'-非翻译区结合以调节其表达。总之，本研究结果表明，miR-455-5p在ESCC组织中的表达降低，且与淋巴转移和分化呈负相关。作为一种肿瘤抑制基因，miR-455-5p通过抑制Rab31的表达来抑制ESCC Eca109细胞的增殖、迁移和侵袭。
