基于网络药理学预测抗癌精方干预结直肠癌的作用及实验验证＊

目的 基于网络药理学采用数据挖掘的方法分析抗癌精方治疗结直肠癌的作用及其作用机制。方法 通过TCMSP数据库检索获得抗癌精方的化学成分,然后根据药代动力学参数筛选出其活性成分,通过Swiss Target Prediction Database对活性成分进行靶点预测;检索TCMNPAS、OMIM、TTD和DrugBank数据库获得结直肠癌的作用基因,二者映射获得抗癌精方治疗结直肠癌的关键活性成分和核心靶点;进一步分析核心靶点对应的生物学功能及相关的信号通路,选取前10条通路所包含的基因与核心靶点进行映射获得与抗癌精方治疗结直肠癌关联程度较高的信号通路;最后通过CCK8实验、流式细胞术测细胞凋亡和Western blot验证抗癌精方对结直肠癌细胞HCT116的抑制作用及其作用机制。结果 研究共筛选出抗癌精方的关键活性成分26个,对应核心靶点145个,取GO分析前10条生物信息,获得信号通路前10条与抗癌精方治疗结直肠癌的关联度排序。细胞水平验证了抗癌精方可以抑制结直肠癌HCT116细胞增殖,升高细胞凋亡率,且呈浓度依赖性;Western blot实验显示抗癌精方可降低p-PI3K、p-AKT、Bcl-2的蛋白表达,升高cleaved caspase3和Bax的蛋白表达,表明抗癌精方具有诱导结直肠癌HCT116细胞凋亡的作用,可能是通过抑制PI3K/AKT信号通路的调控实现,这与网络药理学的分析结果相互印证。结论 本研究创新性的采用网络药理学方法结合实验验证初步探索抗癌精方治疗结直肠癌的作用及其机制,为后续深入的基础研究奠基,为临床应用提供客观支持。

Network-based Pharmacology Prediction and Experimental Validation of Kangai Jingfang Intervention in Colorectal Cancer

Objective To analyze the effect of Kangai Jingfang (KAJF) for colorectal cancer (CRC) and its mechanism based on network pharmacology using data mining method.Methods The chemical composition of KAJF was obtained by searching the TCMSP database, and then the active ingredients were screened according to the pharmacokinetic parameters, and the target prediction of the active ingredients was performed by Swiss Target Prediction Database; the TCMNPAS, OMIM, TTD and DrugBank databases were searched to obtain the action genes of colorectal cancer, and the two were mapped to obtain the key active ingredients and core targets of the anti-cancer essence formula for colorectal cancer. The key active ingredients and core targets of anti-cancer essence formula for colorectal cancer were obtained by searching TCMNPAS, OMIM, TTD and DrugBank databases, and then mapping the two to obtain the key active ingredients and core targets of anti-cancer essence formula for colorectal cancer; further analyzed the biological functions and related signaling pathways of the core targets, and selected the genes contained in the top 10 pathways and the core targets for mapping to obtain the signaling pathways with a higher degree of association with anti-cancer essence formula for colorectal cancer; finally validated by CCK8 assay, apoptosis measurement by flow cytometry and Finally, the inhibitory effect of KAJF on colorectal cancer cells HCT116 and its mechanism of action were verified by CCK8 assay, flow cytometry apoptosis and Western blot.Results A total of 26 key active ingredients of KAJF were screened out, corresponding to 145 core targets, and the top 10 bioinformatics were taken for GO analysis to obtain the correlation ranking of the top 10 signaling pathways with KAJF for colorectal cancer. It was verified at the cellular level that KAJF could inhibit the proliferation and increase the apoptosis rate of colorectal cancer HCT116 cells in a concentration-dependent manner; Western blot showed that Anti-Cancer Essence could decrease the protein expression of p-PI3K, p-AKT and Bcl-2, and increase the protein expression of cleaved caspase3 and Bax, indicating that KAJF could induce apoptosis in colorectal cancer HCT116 cells. The effect of KAJF on inducing apoptosis of HCT116 cells in colorectal cancer may be achieved by inhibiting the regulation of PI3K/AKT signaling pathway, which is corroborated with the results of network pharmacology analysis.Conclusion In this study, an innovative network pharmacology approach combined with experimental validation was used to initially explore the effects and mechanisms of KAJF in the treatment of colorectal cancer, laying the foundation for subsequent in-depth basic research and providing objective support for clinical application.