Effect of evodiamine and berberine on the interaction between DNMTs and target microRNAs during malignant transformation of the colon by TGF-β1
Effect of evodiamine and berberine on the interaction between DNMTs and target microRNAs during malignant transformation of the colon by TGF-β1
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DOI：
10.3892/or.2017.5379
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发表时间：
2017-03-01
期刊：
影响因子：
4.2
通讯作者：
Wen, Bin
中科院分区：
文献类型：
--
作者：
Huang, Chao;Liu, Hong;Wen, Bin
组织微环境在肿瘤的发生和转化生长因子-β1结直肠癌中起着关键作用。微环境中的转化生长因子-β1刺激肿瘤的形成。采用转化生长因子-β1诱导的体外恶性转化模型，观察吴茱萸碱和黄连素对该模型中DNA甲基转移酶(DNMTs)和靶microRNAs(MiRNAs)相互作用的影响。培养7日龄新生大鼠结肠组织，用转化生长因子-β1体外诱导大鼠结肠组织恶性转化48h后，分别给予吴茱萸碱和黄连素作用24 h，苏木精-伊红染色观察组织形态变化，免疫组织化学结合基因芯片分析检测DNMT和我们前期研究中筛选的靶向miRNAs的表达水平，并用实时定量聚合酶链式反应进行定量。转化生长因子-β1作用24 h后，DNMT1、DNMT3A、DNMT3b和miR-152(靶DNMT1)、miR-429(靶DNMT3A)和miR-29a(靶DNMT3A/3B)的表达水平显著降低，而48 h后，DNMT1和DNMT3A的表达水平显著升高，但其靶miRNAs的表达水平仍然下降。经DNMT抑制剂(5-aza-DC)处理后，miRNAs的表达水平有较大程度的提高，但未达到正常水平。黄连素和吴茱萸碱作用24 h后，DNMT1、DNMT3a、Dnmt3b和miR-152、miR-429、miR-29a的表达均增加。综上所述，本研究结果表明，miRNAs也可以受其相应的DNMT转录后调控，并且黄连素和吴茱萸碱调节这些基因的表达，这为预防和治疗结直肠癌提供了早期的表观遗传学证据。
The tissue microenvironment functions as a crucial player in carcinogenesis, and transforming growth factor-beta 1 colorectal cancer. (TGF-beta 1) within the microenvironment stimulates the formation of neoplasms. Using an in vitro model of malignancy induced by TGF-beta 1, we assessed the effect of evodiamine and berberine on the interaction between DNA methyltransferases (DNMTs) and target microRNAs (miRNAs) in the model. Colon tissues from neonatal rats 7 days of age were cultured and malignancy was induced by TGF-beta 1 in vitro for 48 h, and then the tissues were respectively treated with evodiamine and berberine for 24 h. Morphological alteration of tissues was were observed using hematoxylin and eosin staining, and the expression levels of DNMTs and targeted miRNAs screened by bioinformatics software combined with Gene chip analysis in our previous study were detected by immunohistochemistry and quantified by real-time PCR. Twenty-four hours after treatment with TGF-beta 1, expression levels of DNMT1, DNMT3A, DNMT3B and miR-152 (target DNMT1), miR-429 (target DNMT3A) and miR-29a (target DNMT3A/3B) were markedly decreased; however, after 48 h, the expression levels of DNMT1 and DNMT3A were significantly increased, but their target miRNAs were still decreased. After treatment with a DNMT inhibitor (5-Aza-dC), expression levels of the miRNAs were increased to a larger extent, but did not reach normal levels. After treatment with berberine and evodiamine for 24 h, respectively, increased expression of DNMT1, DNMT3A, DNMT3B and miR-152, miR-429, miR-29a was noted. In conclusion, the results of the present study suggest that miRNAs can also be post-transcriptionally regulated by their corresponding DNMTs and that berberine and evodiamine regulate the expression of these genes, which provides early epigenetic evidence for the prevention and therapy of colorectal cancer.