基于网络药理学与细胞实验探索藏药十一味草果丸治疗结直肠癌机制

To explore the mechanism of Tibetan medicine Eleven Caoguo pills in the treatment of colorectal cancer based on network pharmacology and cell experiments

目的通过生物信息学挖掘结直肠癌相关基因,基于网络药理学对十一味草果丸治疗结直肠癌的机制进行探索研究,利用HCT116结直肠癌细胞进行实验对网络药理学预测结果进行验证。方法结直肠癌疾病相关基因,通过GEO数据库、孟德尔人类遗传基因库、GeneCard数据库获取;基于知网、TCMSP数据库对藏药十一味草果丸主要成分进行检索,探索其相关靶点蛋白通过Uniport数据库与DrugeBank数据库获取蛋白质对应基因;使用GeneCards数据库对结直肠癌疾病相关的所有基因与筛选得到的十一味草果丸作用的靶基因进行比对选取相关性高的基因用以探索二者间的相关关系;使用DAVID数据库对得到的相关基因进行富集分析,得到相关的生物过程与信号通路;使用分子对接技术对活性成分进行研究。最后利用HCT116结直肠癌细胞对网络药理学预测的十一味草果丸作用于结直肠癌的效果进行检验。结果GEO数据库筛选后获得差异基因与GeneCard对比后与疾病相关度大于30分的相关基因288个。数据库检索得到药物有效化学成分169个,对化学成分进行预测靶点蛋白进行检索,去重后得到508个靶基因。508个药物基因与288个疾病相关基因取交集后64个共同基因。KEGG富集结果主要有癌症通路、PI3K-Akt信号通路、P53信号通路等。通过使用水和乙酸乙酯提取十一味草果丸中的药物成分,利用水和二甲基亚砜(DMSO)进行溶解并控制其浓度后,对结直肠癌HCT116细胞进行干预,其中5氟尿嘧啶(5-FU)作阳性对照而提取药物设置不同浓度剂量组干预结果显示,乙酸乙酯提取的药物作用效果优于水提取药物和5-FU。结论藏药十一味草果丸可以通过多个靶点和多条通路对结直肠癌细胞起到抑制和促进凋亡作用。

Objective Through bioinformatics mining of colorectal cancer related genes,exploratory study on the mechanism of eleven flavour grass fruit pills in treating colorectal cancer based on cyberpharmacology,and experimental verification of cyberpharmacology prediction results using HCT116 colorectal cancer cells.Methods The genes related to colorectal cancer were obtained from the GEO database,Mendelian Human Genome Database and GeneCard database;based on the Knowledge Network and TCMSP database,we searched the main components of the Tibetan medicine Shodaiyi Caojiao Pill and explored the target proteins through the Uniport and DrugeBank databases to obtain the corresponding genes of the proteins;we compared all the genes related to colorectal cancer with the target genes obtained from the screening of the Shodaiyi Caojiao Pill by GeneCards database.The GeneCards database was used to compare all the genes related to colorectal cancer with the target genes obtained from the screening of the Eleven Wisdom Grass Fruit Pill,and the genes with high correlation were selected to explore the correlation between the two;the DAVID database was used to analyse the enriched genes,and to obtain the related biological processes and signaling pathways;the molecular docking technique was used to study the active ingredients.Finally,HCT116 colorectal cancer cells were used to test the network pharmacological prediction of the effect of eleven flavours of grass jelly pill on colorectal cancer.Results The GEO database was screened to obtain 288 related genes with a correlation score of greater than 30 with the disease after the differential gene was compared with GeneCard.The database search yielded 169 active chemical ingredients,and the predicted target proteins of the chemical ingredients were searched,and 508 target genes were obtained after de-emphasis.508 drug genes were intersected with 288 disease-related genes and 64 common genes were obtained.KEGG enrichment results mainly included Pathways in cancer,PI3K-Akt signalling pathway,P53 signalling pathwayand so on.By using water and ethyl acetate to extract the drug components in the eleven flavour grass fruit pill,using water and dimethyl sulfoxide(DMSO)to dissolve and control its content,after the intervention of colorectal cancer HCT116 cells,in which 5-fluorouracil(5-FU)as a positive control while the extracted drug set up different concentrations of the dose groups of the intervention results showed that ethyl acetate extraction of the drug effect is better than the water extracted drug and 5-FU.The results showed that the effect of ethyl acetate extract was better than that of water extract and 5-FU.Conclusion The Tibetan medicine eleven flavour herbs and fruits pills can inhibit and promote apoptosis of colorectal cancer cells through multiple targets and multiple pathways.