天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (1): 19-24.doi: 10.11958/20192289

• 细胞与分子生物学 • 上一篇    下一篇

miR-149抑制结直肠癌细胞迁移的分子机制研究

刘晓柱，李银凤   

  1. 基金项目：贵州省科技计划项目（黔科合基础［2018］1069） 作者单位：贵州理工学院食品药品制造工程学院（邮编550003） 作者简介：刘晓柱（1984），男，博士，副教授，主要从事肿瘤分子生物学方向基础研究
  • 收稿日期:2019-07-26 修回日期:2019-10-23 出版日期:2020-01-15 发布日期:2020-01-15
  • 通讯作者: 刘晓柱 E-mail:liuxiaozhu_840914@163.com
  • 基金资助:
    miR-149 激活 Wnt/β-catenin 信号通路 调控结直肠癌侵袭与转移的分子机制研究

The molecular mechanism of inhibiting effect of miR-149 on the metastasis of human colonic carcinoma cells

LIU Xiao-zhu, LI Yin-feng   

  1. College of Food & Pharmaceutical Engineering, Guizhou Institute of Technology, Guiyang 550003, China
  • Received:2019-07-26 Revised:2019-10-23 Published:2020-01-15 Online:2020-01-15

PDF (PC)

5767

摘要： 目的 探讨miR-149对结直肠癌（CRC）细胞增殖、侵袭、迁移及凋亡的影响及其作用机制。方法 实时荧 光定量PCR（q-PCR）方法检测miR-149在CRC细胞SW620、LS174T和人结肠上皮细胞FHC中的表达水平；荧光素酶 分析法验证 STAT3 与 miR-149 之间的靶向结合关系；q-PCR 和 Western blot 检测 miR-149 过表达对 CRC 细胞中 STAT3、p-STAT3 mRNA和蛋白表达的影响。将miR-149 mimics与STAT3过表达质粒单独或者共转染至CRC细胞 中，并分为miR-NC组、miR-149 mimics组、miR-149 mimics+pEGFP/STAT3组。采用CCK-8法、Transwell法、细胞划 痕法、流式细胞术分别检测各分组CRC细胞的增殖、侵袭、迁移和凋亡情况。结果 与正常结肠上皮细胞FHC相比， CRC 细胞 SW620、LS174T 中 miR-149 表达水平受到抑制（P＜0.01）。荧光素酶实验证实，在 CRC 细胞中 STAT3 是 miR-149的一个直接靶基因。过量表达miR-149抑制STAT3、p-STAT3 mRNA和蛋白的表达，降低CRC细胞增殖能 力、侵袭能力以及迁移能力（P＜0.01）。同时，过量表达 miR-149 也可促进 CRC 细胞的凋亡（P＜0.01）。但过表达 STAT3可解除miR-149对CRC细胞增殖、侵袭以及迁移能力的抑制作用。结论 miR-149通过靶向STAT3抑制结直 肠癌细胞的增殖、侵袭与迁移，同时促进细胞的凋亡。

关键词: 结直肠肿瘤, 微RNAs, STAT3转录因子, 细胞增殖, 细胞运动, 细胞凋亡, miR-149

Abstract: Objective To investigate the regulation and function of miR-149 in human colorectal cancer cell lines. Methods miR-149 expression patterns were detected in SW620 and LS174T cell lines using quantitative real-time fluorescence PCR (q-PCR) method. And then, the target gene of miR-149 was explored via luciferase reporter assay. The expressions of STAT3 and p-STAT3 mRNA and proteins in CRC cells were detected by q-PCR and Western blot assay, respectively. miR-149 mimics and pEGFP/STAT3 vector were transfected or co-transformation into CRC cells. CRC cells were divided into miR-NC group, miR-149 mimics group and miR-149 mimics + pEGFP / STAT3 group. Proliferation, migration, invasion and apoptosis were determined by CCK-8 assay, wound-healing assay, transwell assay and flow cytometry, respectively. Results miR-149 expression was down-regulated in SW620 and LS174T cell lines compared to that of the normal colon epithelial cell FHC detected using q-PCR methods (P＜0.01). Then, STAT3 was identified as a direct target gene of miR-149 in CRC cells by luciferase reporter assay. Further studies indicated that the introduction of miR-149 was able to down-regulate the mRNA and proteins expression of STAT3 and p-STAT3, suppress cell proliferation, migration and invasion, and promote apoptosis of CRC cells (P＜0.01). However, the over-expression of STAT3 could decrease the inhibiting effect of miR-149 on the proliferation,migration and invasion of CRC cells. Conclusion miR-149 can inhibit proliferation, invasion and migration and promote apoptosis of CRC cells via targeting STAT3.

Key words: colorectal neoplasms, microRNAs, STAT3 transcription factor, cell proliferation, cell movement, apoptosis, miR-149