Li Chunyan, Lu Shuming, Wang Yingyan, Guo Shibin, Zhao Tianyu, Wang Xiaosheng, Song Bo
Department of Gastroenterology, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China.
Laboratory Center for Diagnostics, Dalian Medical University, Dalian, Liaoning 116044, P.R. China.
Mol Med Rep. 2017 Feb;15(2):833-838. doi: 10.3892/mmr.2016.6088. Epub 2016 Dec 29.
The aim of the present study was to investigate the role of microRNA (miR)‑34a expression in the proliferation, invasion and metastasis of colon cancer and its underlying mechanisms. HCT116 cells were cultured in high‑sugar Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum and 1000 U/ml penicillin‑streptomycin. Following digestion and resuspension, the cells were used for transfection, expression and western blot analysis. HCT116 cells from miR‑34a transfection, negative control and blank control groups were seeded into a 96‑well plate at a density of 105 cells/ml, and 200 µl complete DMEM was added. The data are presented as the mean ± standard error. A one‑way analysis of variance was performed to compare groups. miR‑34a‑HCT116 cells demonstrated significantly increased expression levels of miR‑34a. The proliferation of HCT116 cells with overexpression of miR‑34a was significantly inhibited to 0.49±0.11 compared with the blank control group (P<0.001). Compared with the blank control and negative control groups, the protein expression levels of B‑cell lymphoma 2 (Bcl‑2) were markedly reduced in the miR‑34a transfected group. Furthermore, the protein expression levels of Bcl‑2‑associated X protein were significantly increased and those of matrix metalloproteinase (MMP)‑2 and MMP‑9 were markedly reduced in the miR‑34a transfected group, MMP‑9 to a greater extent. The present study suggested that overexpression of miR‑34a may inhibit the proliferation, invasion and metastasis of HCT116 cells.
本研究旨在探讨微小RNA（miR）-34a表达在结肠癌增殖、侵袭和转移中的作用及其潜在机制。将HCT116细胞培养于含10%胎牛血清和1000 U/ml青霉素-链霉素的高糖杜氏改良 Eagle 培养基（DMEM）中。消化并重悬后，将细胞用于转染、表达及蛋白质印迹分析。将来自miR-34a转染组、阴性对照组和空白对照组的HCT116细胞以105个细胞/ml的密度接种到96孔板中，并加入200 μl完全DMEM。数据以平均值±标准误差表示。采用单因素方差分析比较各组。miR-34a-HCT116细胞显示miR-34a表达水平显著升高。与空白对照组相比，miR-34a过表达的HCT116细胞增殖受到显著抑制，降至0.49±0.11（P<0.001）。与空白对照组和阴性对照组相比，miR-34a转染组中B细胞淋巴瘤2（Bcl-2）的蛋白表达水平明显降低。此外，miR-34a转染组中Bcl-2相关X蛋白的蛋白表达水平显著升高，基质金属蛋白酶（MMP）-2和MMP-9的蛋白表达水平明显降低，MMP-9降低幅度更大。本研究表明，miR-34a过表达可能抑制HCT116细胞的增殖、侵袭和转移。
