miRNA-200b 通过靶向 SMYD2/p53 信号通路对肝癌细胞增殖的影响。
Effect of miRNA-200b on the proliferation of liver cancer cells via targeting SMYD2/p53 signaling pathway.
机构信息
Department of Pharmacy, Third Xiangya Hospital, Central South University, Changsha 410013.
Department of Pharmacy, Women and Children's Health Care Hospital of Linyi, Linyi Shandong 276000, China.
出版信息
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2022 Oct 28;47(10):1303-1314. doi: 10.11817/j.issn.1672-7347.2022.210521.
OBJECTIVES
Our previous study has verified that high level of SET and MYND domain-containing protein 2 (SMYD2) plays an important role in acquiring aggressive ability for liver cancer cells in hepatocellular carcinoma. MiR-200b as a tumor suppressor gene involves in a variety of cancers. This study aims to investigate the correlation between miR-200b and SMYD2 in hepatocellular carcinoma and the underlying mechanism.
METHODS
Firstly, the levels of SMYD2 and miR-200b in hepatocellular carcinoma tissues and matched adjacent non-tumor liver tissues were tested with real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. Secondly, we evaluated the interaction between miR-200b and SMYD2 using dual-luciferase reporter assay. Thirdly, we elucidated the effect of miR-200b on SMYD2 and its downstream targets p53/CyclinE1. Finally, we silenced SMYD2 in hepatocellular carcinoma cell lines to investigate its effect on tumor proliferation and cell cycle progression, and further confirmed the correlation among SMYD2 and p53/CyclinE1.
RESULTS
Compared with the matched adjacent non-tumor liver tissues, miR-200b was obviously decreased, and SMYD2 was significantly increased in hepatocellular carcinoma (both <0.05). Spearman's rank correlation revealed that miR-200b expression was negatively correlated with SMYD2 (<0.01). Computer algorithm and dual-luciferase reporter assay revealed that miR-200b directly targeted and suppressed SMYD2 in HEK 293T cells. The down-regulated miR-200b expression promoted hepatoma cell proliferation (<0.05) and increased SMYD2 expression(<0.01), while the up-regulated expression of miR-200b had an opposite effect. The knockdown of SMYD2 suppressed the proliferation of MHCC-97L cells (<0.01), down-regulated CyclinE1, and up-regulated p53 expression (both <0.05).
CONCLUSIONS
MiR-200b is involved in hepatocellular carcinoma progression via targeting SMYD2 and regulating SMYD2/p53/CyclinE1 signaling pathway and may be used as a potential target for hepatocellular carcinoma treatment.
目的
我们之前的研究已经证实，SET 和 MYND 结构域包含蛋白 2（SMYD2）的高水平在肝癌细胞获得侵袭能力中发挥重要作用。miR-200b 作为一种肿瘤抑制基因，涉及多种癌症。本研究旨在探讨 miR-200b 与肝癌中的 SMYD2 之间的相关性及其潜在机制。
方法
首先，采用实时逆转录聚合酶链反应（RT-PCR）和 Western blot 检测肝癌组织及配对的癌旁非肿瘤组织中 SMYD2 和 miR-200b 的水平。其次，采用双荧光素酶报告基因检测评估 miR-200b 与 SMYD2 之间的相互作用。再次，阐明 miR-200b 对 SMYD2 及其下游靶基因 p53/CyclinE1 的影响。最后，沉默肝癌细胞系中的 SMYD2，探讨其对肿瘤增殖和细胞周期进程的影响，并进一步证实 SMYD2 与 p53/CyclinE1 之间的相关性。
结果
与配对的癌旁非肿瘤组织相比，miR-200b 在肝癌组织中明显下调，SMYD2 明显上调（均<0.05）。Spearman 秩相关分析显示，miR-200b 的表达与 SMYD2 呈负相关（<0.01）。计算机算法和双荧光素酶报告基因检测显示，miR-200b 可在 HEK 293T 细胞中直接靶向并抑制 SMYD2。下调 miR-200b 的表达促进肝癌细胞增殖（<0.05）并增加 SMYD2 的表达（<0.01），而上调 miR-200b 的表达则产生相反的效果。SMYD2 的敲低抑制 MHCC-97L 细胞的增殖（<0.01），下调 CyclinE1，并上调 p53 的表达（均<0.05）。
结论
miR-200b 通过靶向 SMYD2 及其调节 SMYD2/p53/CyclinE1 信号通路参与肝癌的进展，可能作为肝癌治疗的潜在靶点。
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