MDA-9/Syntenin-NF-κB-RKIP 环路在人肝癌中的潜在治疗应用。
Potential Therapeutic Applications of MDA-9/Syntenin-NF-κB-RKIP Loop in Human Liver Carcinoma.
机构信息
Department of Biological, Chemical and Pharmaceutical Sciences and Technologies, School of Science, University of Palermo, Italy.
Department of Health Sciences and Mother and Child Care "G. D'Alessandro", School of Medicine, University of Palermo, Italy.
出版信息
Curr Mol Med. 2018;18(9):630-639. doi: 10.2174/1566524019666190104105043.
BACKGROUND
Overexpression of MDA-9/Syntenin occurs in multiple human cancer cell lines and is associated with higher grade of tumor classification, invasiveness and metastasis. In some cases, its role in cancer biology depends on relationships between MDA-9/Syntenin and NF-κB.
OBJECTIVE
This study aims to analyze the presence of a regulation loop like that between MDA-9/Syntenin - NF-κB - RKIP in human liver carcinoma.
METHODS
Transient transfection was performed with siRNA anti-MDA-9/Syntenin. Expression of different factors was evaluated by Real time-PCR and Western blotting, while NF-κB activation by TransAM assay. Invasion capacity was analyzed by Matrigel Invasion Assay and the effects of agents on cell viability were examined by MTS assay.
RESULTS
We have examined basal expression of MDA-9/Syntenin in three cell lines of human liver carcinoma (HA22T/VGH, Hep3B and HepG2). In all cell lines there was an inverse relationship between MDA-9/Syntenin and RKIP expression levels, and a positive correlation between MDA-9/Syntenin expression and NF-κB activation levels. By silencing with a siRNA anti-MDA-9/Syntenin we observed in all cell lines a very strong increase of RKIP at mRNA level. Interestingly, in all cell lines, inhibition of MDA- 9/Syntenin expression induced NF-κB downregulation and contemporary a reduction in invasion ability MMP-2 dependent. Finally, we showed a good additive effect of MDA- 9/Syntenin siRNA when associated with Curcumin or Doxorubicin on cell growth inhibition.
CONCLUSION
Our data confirm the key role of MDA-9/Syntenin in HCC biology. The presence of a regulation loop among MDA-9/Syntenin, NF-κB and RKIP provide new pharmacological approaches.
背景
MDA-9/Syntenin 在多种人类癌细胞系中过表达，与肿瘤分级、侵袭性和转移率较高相关。在某些情况下，它在癌症生物学中的作用取决于 MDA-9/Syntenin 与 NF-κB 之间的关系。
目的
本研究旨在分析 MDA-9/Syntenin-NF-κB-RKIP 之间的调控环路是否存在于人类肝癌中。
方法
采用 siRNA 对 MDA-9/Syntenin 进行瞬时转染。通过实时 PCR 和 Western blot 评估不同因子的表达情况，通过 TransAM 测定 NF-κB 的激活情况。通过 Matrigel 侵袭实验分析侵袭能力，通过 MTS 实验检测药物对细胞活力的影响。
结果
我们已经检测了三种人肝癌细胞系（HA22T/VGH、Hep3B 和 HepG2）中 MDA-9/Syntenin 的基础表达。在所有细胞系中，MDA-9/Syntenin 与 RKIP 表达水平呈负相关，与 NF-κB 激活水平呈正相关。通过 siRNA 沉默 MDA-9/Syntenin，我们观察到在所有细胞系中，RKIP 的 mRNA 水平都有非常显著的增加。有趣的是，在所有细胞系中，抑制 MDA-9/Syntenin 的表达诱导了 NF-κB 的下调，同时 MMP-2 依赖的侵袭能力也降低。最后，我们发现 MDA-9/Syntenin siRNA 与姜黄素或阿霉素联合使用时对细胞生长抑制有良好的协同作用。
结论
我们的数据证实了 MDA-9/Syntenin 在 HCC 生物学中的关键作用。MDA-9/Syntenin、NF-κB 和 RKIP 之间存在调控环路，为新的药物治疗方法提供了依据。
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