基于网络药理学技术探讨化痰散结方对阿霉素耐药乳腺癌PI3K-Akt-mTOR信号通路相关蛋白的影响

Effects of Huatan Sanjie Recipe on PI3K-Akt-mTOR Signaling Pathway Protein in Adriamycin-resistant Breast Cancer under Guidance of Network Pharmacology

目的：基于网络药理学建立多柔比星/阿霉素耐药相关的蛋白质相互作用网络并通体外实验研究探究化痰散结方逆转耐药的相关分子学机制。方法：筛选pubmed中人类OMMIM遗传数据库的乳腺癌多柔比星/阿霉素耐药相关基因,应用STRING软件进行文本挖掘并建立蛋白质相互作用网络;将互作数据读入Cytoscape3.4.0,分别应用插件JEPETTO 1.3.1和MCODE1.4.2实现拓扑分析和聚类分析;使用Cytoscape中的插件Clusterviz进一步挖掘网络中可能包含的分子复合物;将所得复合物导入DAVID,富集其中的分子复合物通路。针采用MTT法、Western Blot法对细胞及关键通路进行体外实验研究,进而观察化痰散结方对MCF-7/ADR细胞增殖及相关信号通路表达的影响。结果：乳腺癌多柔比星/阿霉素耐药蛋白质网络是一个多基因构成的网络并且由多信号通路协同发挥调节作用,并非简单由单基因或信号通路调控;在所得相互作用网络中,PI3K-Akt-mTOR很可能作为＂关键节点＂发挥调控作用;化痰散结方含药血清能够抑制MCF-7/ADR细胞的增值,其内在分子学机制可能与抑制PI3K-Akt-mTOR通路的表达有关。结论：化痰散结方能够逆转人乳腺癌多柔比星/阿霉素耐药,其耐药相关蛋白质相互作用网络复杂,其中PI3K-Akt-mTOR很可能是其中的关键调控点,而化痰散结方抑制MCF-7/ADR细胞增殖的作用机制之一是抑制PI3K-Akt-mTOR通路的表达。

Objective： To establish a drug-related network of doxorubicin/adriamycin resistance based on network pharmacology and explore the molecular mechanism of reversal of drug resistance in Huatan Sanjie Recipe. Methods： We screened breast cancer doxorubicin/adriamycin-resistant genes in pubmed human OMMIM genetic database and used STRING software for text mining and establishment of protein interaction network. The interaction data was put into Cytoscape 3. 4. 0,using plug-ins JEPETTO 1. 3. 1 and MCODE 1. 4. 2 respectively to achieve topology analysis and clustering analysis. And then we used plugins in Cytoscape Clusterviz to further dig out possible molecular complexes in the network. The complexes were put into online DAVID,enriching the molecular complex pathway. For critical access,MTT and Western blot were used to study the vitro experimental study of cytology. And we observed the effect of Huatan Sanjie Recipe on the proliferation of MCF-7/ADR cells and the expression of related signaling pathway. Results： Breast cancer doxorubicin/adriamycin-resistant protein network is a multi-gene network and is coordinated by multiple signaling pathways. PI3 K-Akt-mTOR is likely to be one of the key control points. One of the mechanisms of action of Huatan Sanjie Recipe on MCF-7/ADR cell proliferation is the expression of PI3 K-Akt-mTOR pathway. Conclusion：Huatan Sanjie Recipe can reverse the drug resistance of breast cancer doxorubicin/adriamycin and the related proteins are complex. PI3 K-Akt-mTOR may be the key point. Huatan Sanjie Recipe can inhibit MCF-7/ADR cells proliferation by inhibiting PI3 K-Akt-mTOR expressions.