微小RNA-196b-5p通过靶向ING5促进结直肠癌的恶性进展。
MicroRNA-196b-5p promotes malignant progression of colorectal cancer by targeting ING5.
作者信息
Xin He, Wang Chuanzhuo, Chi Yuan, Liu Zhaoyu
机构信息
Department of Radiology, Shengjing Hospital of China Medical University, 36 Sanhao Street, Shenyang, 110004 People's Republic of China.
出版信息
Cancer Cell Int. 2020 Apr 10;20:119. doi: 10.1186/s12935-020-01200-3. eCollection 2020.
BACKGROUND
miR-196b-5p expression is deregulated in many malignant tumors. Although miR-196b-5p has been implicated in the malignant transformation of colorectal cancer, its role in this specific type of cancer has not been fully explored. Thus, the present study was aimed to examine the cellular function of miR-196b-5p and its role in malignant biological behavior in colorectal cancer.
METHODS
miR-196b-5p expression was measured in colorectal cancer tissues and cell lines using quantitative real-time PCR. Cell counting kit-8 (CCK-8) assay and Transwell assay were used to detect proliferation, migration, and invasion in cell lines, whereas flow cytometry was applied to study apoptosis. Western blot analysis was performed to measure the protein levels. Dual luciferase reporter assay was used to investigate the interaction between miR-196b-5p and ING5. Tumor formation was evaluated in mice.
RESULTS
MiR-196b-5p was abundantly expressed in colorectal cancer tissues and cell lines, whereas ING5 was expressed at low levels. MiR-196b-5p was successfully overexpressed or knocked down in colorectal cancer cells. We found that miR-196b-5p overexpression significantly accelerated the proliferation, cell cycle, migration and invasion, while inhibited cell apoptosis in colorectal cancer cells. However, miR-196b-5p inhibitor showed the opposite effects. Moreover, ING5 overexpression or knockdown was successfully performed in colorectal cancer cells. ING5 overexpression suppressed proliferation, migration, invasion, the phosphorylation of PI3K, Akt as well as MEK, and promoted cell apoptosis, which could be reversed by ING5 knockdown. Additionally, ING5 was identified as a target of miR-196b-5p through bioinformatics analysis and a luciferase activity assay. Furthermore, ING5 knockdown could attenuate the decrease in proliferation, migration, invasion, and the protein levels of p-PI3K, p-Akt, and p-MEK, which were induced by miRNA-196b-5p inhibitor. Besides, miR-196b-5p knockdown inhibited tumor growth, whereas ING5 knockdown elevated it in vivo.
CONCLUSIONS
In conclusion, miR-196b-5p promotes cell proliferation, migration, invasion, and inhibits apoptosis in colorectal cancer by targeting ING5.
背景
miR-196b-5p在许多恶性肿瘤中表达失调。尽管miR-196b-5p与结直肠癌的恶性转化有关，但其在这种特定类型癌症中的作用尚未得到充分研究。因此，本研究旨在探讨miR-196b-5p的细胞功能及其在结直肠癌恶性生物学行为中的作用。
方法
采用定量实时PCR检测结直肠癌组织和细胞系中miR-196b-5p的表达。使用细胞计数试剂盒-8（CCK-8）检测和Transwell检测来检测细胞系中的增殖、迁移和侵袭，而流式细胞术用于研究细胞凋亡。进行蛋白质印迹分析以测量蛋白质水平。使用双荧光素酶报告基因检测来研究miR-196b-5p与ING5之间的相互作用。在小鼠中评估肿瘤形成。
结果
miR-196b-5p在结直肠癌组织和细胞系中大量表达，而ING5表达水平较低。miR-196b-5p在结直肠癌细胞中成功过表达或敲低。我们发现miR-196b-5p过表达显著加速了结直肠癌细胞的增殖、细胞周期、迁移和侵袭，同时抑制了细胞凋亡。然而，miR-196b-5p抑制剂显示出相反的效果。此外，ING5在结直肠癌细胞中成功过表达或敲低。ING5过表达抑制了增殖、迁移、侵袭、PI3K、Akt以及MEK的磷酸化，并促进了细胞凋亡，ING5敲低可使其逆转。此外，通过生物信息学分析和荧光素酶活性检测确定ING5是miR-196b-5p的靶标。此外，ING5敲低可减弱由miRNA-196b-5p抑制剂诱导的增殖、迁移、侵袭以及p-PI3K、p-Akt和p-MEK蛋白水平的降低。此外，miR-196b-5p敲低抑制了肿瘤生长，而ING5敲低在体内则使其升高。
结论
总之，miR-196b-5p通过靶向ING5促进结直肠癌细胞的增殖、迁移、侵袭并抑制细胞凋亡。
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