培替布林通过下调 E6/E7-Pi3k/Akt 通路诱导宫颈癌细胞凋亡：一种网络药理学研究。

Paiteling induces apoptosis of cervical cancer cells by down-regulation of the E6/E7-Pi3k/Akt pathway: A network pharmacology.

机构信息

School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China.

出版信息

J Ethnopharmacol. 2023 Apr 6;305:116062. doi: 10.1016/j.jep.2022.116062. Epub 2022 Dec 17.

DOI:10.1016/j.jep.2022.116062

PMID:36535331

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Human papillomavirus (HPV) infection is considered to be the main pathogen causing intraepithelial neoplasia. Paiteling (PTL) has been used to treat intraepithelial neoplasia caused by human papillomavirus (HPV) infection for more than 20 years in China, but its specific mechanism of action is not very clear, and further research is still needed.

OBJECTIVE

This study designed a comprehensive strategy to study the pharmacological mechanism of paiteling in regulating cervical cancer cell apoptosis by integrating LC-MS/MS, network pharmacology and pharmacological experiments.

METHODS

We used liquid chromatography-tandem mass spectrometry to detect the active substances in PTL and performed protein-protein interaction analysis on the intersection of the targets of these key compounds and the targets of intraepithelial neoplasia. Additionally, by using Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes (KEGG), the potential pathway of PTL against HPV-induced intraepithelial neoplasia was predicted. Finally, we used HeLa and Ect1/E6E7 cells for experimental verification.

RESULTS

The protein-protein interaction network predicted that AKT1, TP53, MYC, STAT3, MTOR, and MAPK were pivotal targets for PTL to inhibit epithelial neoplasia. KEGG enrichment analysis showed that the Pi3k/Akt pathway and HPV infection had scientific significance. Compared to the control group, after PTL diluent stimulated HeLa and Ect1/E6E7 cells for 24 h, cell viability, migration, and invasion capabilities were significantly reduced, and cell apoptosis was significantly increased, conforming to a dose-effect relationship and time-effect relationship. PCR, cellular immunohistochemistry, and western blot experiments showed that PTL reduced the expression of E6, Pi3k, E7, Akt, Bcl-xl, while increasing the expression of Bad in HeLa and Ect1/E6E7 cells.

CONCLUSION

PTL can induce cervical cancer cell apoptosis by inhibiting the E6/E7-Pi3k/Akt signaling pathway. It may provide an effective alternative strategy of traditional Chinese medicine for the treatment of epithelial neoplasia caused by HPV infection.

摘要

民族药理学相关性

人乳头瘤病毒（HPV）感染被认为是导致上皮内瘤变的主要病原体。在中国，派特灵（PTL）已经用于治疗 HPV 感染引起的上皮内瘤变 20 多年，但具体作用机制尚不清楚，仍需进一步研究。

目的

本研究通过整合 LC-MS/MS、网络药理学和药理学实验，设计了一种综合策略来研究派特灵调节宫颈癌细胞凋亡的药理机制。

方法

我们使用液相色谱-串联质谱法检测 PTL 中的活性物质，并对这些关键化合物的靶点与上皮内瘤变靶点的交集进行蛋白质-蛋白质相互作用分析。此外，通过使用基因本体论和京都基因与基因组百科全书（KEGG），预测 PTL 针对 HPV 诱导的上皮内瘤变的潜在途径。最后，我们使用 HeLa 和 Ect1/E6E7 细胞进行实验验证。

结果

蛋白质-蛋白质相互作用网络预测 AKT1、TP53、MYC、STAT3、MTOR 和 MAPK 是 PTL 抑制上皮瘤变的关键靶点。KEGG 富集分析表明，Pi3k/Akt 通路和 HPV 感染具有科学意义。与对照组相比，PTL 稀释剂刺激 HeLa 和 Ect1/E6E7 细胞 24 小时后，细胞活力、迁移和侵袭能力明显降低，细胞凋亡明显增加，呈剂量-效应关系和时间-效应关系。PCR、细胞免疫组化和 Western blot 实验表明，PTL 降低了 HeLa 和 Ect1/E6E7 细胞中 E6、Pi3k、E7、Akt、Bcl-xl 的表达，同时增加了 Bad 的表达。