Exploring The Risk Factors Of Primary Liver Cancer And Mining Potential Therapeutic Traditional Chinese Medicine Based On TSMR And CADD

ObjectivePrimary liver cancer is one of the common malignant tumors in the world,with high incidence rate and high mortality.This study will discuss a series of issues based on the etiology,syndrome differentiation,treatment and prognosis of traditional Chinese medicine in treating primary liver cancer,including the causal relationship between primary liver cancer and the internal causes of traditional Chinese medicine,such as "improper diet" and"emotional anger",summarizing the TCM syndrome types of primary liver cancer,and constructing the endoplasmic reticulum immune prognosis model,To explore the prognosis of primary liver cancer,obtain endoplasmic reticulum related genes,use computer aided drug design research methods,and combine reverse network pharmacology to mine potential Chinese medicines for treatment of primary liver cancer,so as to provide leading reference for subsequent drug research and development.Methods1.Mendelian randomized analysis of two samples:through searching relevant databases,genome-wide association study data related to improper diet(smoking,drinking,type 2 diabetes,body mass index,low-density lipoprotein cholesterol,food or drug induced transaminase abnormalities)and smoldering injury to liver(depression)were obtained as exposure factors in this study.The genome-wide association study data of primary liver cancer were selected as outcome factors,and the causal relationship between life factors and primary liver cancer was analyzed by inverse variance weighting,weighted median method,MR Egger regression analysis,simple model method and weighted model.2.By searching relevant Chinese journal databases,screening and obtaining cases and syndrome analysis of primary liver cancer,and disassembling disease specific syndrome elements and disease location syndrome elements,a disease specific disease location database is constructed.Frequency statistics are used to summarize the highest frequency disease specific disease,disease location syndrome elements,and the most common disease specific disease location syndrome element combinations.The obtained database data is imported into the mathematical attribute partial order principle software,Generate relevant attribute partial order diagrams to analyze the maximum commonality of syndrome elements in primary liver cancer,as well as the most common and frequent disease location disease combination.3.Download RNA seq data,tumor mutation data and clinical data of hepatocellular carcinoma and its matched paracancerous samples through GDC database,construct relevant prognosis models through single factor Cox regression analysis and Kaplan Meier survival analysis,and carry out analysis methods including survival curve,survival status assessment,independent prognosis analysis and ROC to verify the independence and robustness of the model,The Gene Ontology database was used to analyze the cell function,and the Kyoto Encyclopedia of Genes and Genomes and the gene probe enrichment analysis database were used to analyze the gene enrichment pathway.Analyze the survival rate of tumor mutation load related data and tumor microenvironment,extract relevant immune genes,conduct survival analysis related to immune cells,and conduct differential analysis on immune checkpoints.3.Based on the above methods,obtain gene targets for primary liver cancer,merge them with other disease database targets,construct relevant PPI networks,conduct GO and KEGG enrichment analysis on relevant targets,obtain enriched primary liver cancer disease pathways and genes,mine traditional Chinese medicine and chemical components through the ITCM database,and use molecular docking methods,Molecular dynamics simulation verifies whether chemical components can effectively dock with disease targets,ultimately obtaining effective compounds and related traditional Chinese medicine.Results1.Mendelian randomization analysis results of improper diet,depression and anger causing liver injury,and primary liver cancer:The above seven exposure factors,except for low-density lipoprotein cholesterol and liver enzyme levels,do not have a direct causal relationship with primary liver cancer.Low density lipoprotein cholesterol and primary liver cancer:IVW(OR=0.422,95%CI:0.269-0.663,P=0.0001),MREgger(OR=0.583,95%CI:0.286-1.18,P=0.153)Weighted median(OR=0.330,95%CI:0.167-0.653,P=0.001),simple mode(OR=0.262,95%CI:0.079-0.866,P=0.039),weighted mode(OR=0.253,95%CI:0.088-0.729,P=0.018);Liver enzyme levels and primary liver cancer:IVW(OR=6.661261e+03,95%CI:2.639747e+02-168093.4,P=9.023504e-08),MREgger(OR=1.908666e+07,95%CI:5.483400e+04-6643697550.1,P=5.728456e-08),weighted median(OR=1.654186e+03,95%CI:1.194529e+01-229072.0,P=3.219623e-03),simple mode(OR=5.794e-01,95%CI:8.131904e-07-412652.0,P=9.367793e-01)Weighted mode(OR=6.0801 17e+01,95%CI:1.908463e-03-1937046.8,P=4.382965e-01).2.Through screening and reading the documents of primary liver cancer syndrome elements,we found 41 literature related to primary liver cancer syndrome elements,involving a total of 19021 cases of primary liver cancer.After standardizing the naming and dismantling of syndrome types of these cases,a total of 41 syndrome types,6 disease location syndrome elements,and 11 disease type syndrome elements were obtained.Among them,disease location syndrome elements were involved 17410 times,and disease type syndrome elements were involved 36379 times.The highest frequency of syndrome types was found in 4173 cases(21.94%)of liver depression and spleen deficiency syndrome.The highest frequency of disease location syndrome elements involved the liver and spleen,accounting for a total of 78.94%.The highest frequency of disease type syndrome elements was found in qi deficiency(25.20%).The highest frequency of combination of pathogenic syndrome elements is Qi stagnation Qi deficiency(28.45%).Through the analysis of attribute partial order principle on the syndrome elements of primary liver cancer,it was found that the most common syndrome elements are liver,qi deficiency,blood stasis,spleen,and qi stagnation,which are the core syndrome elements of primary liver cancer.From the perspective of branch analysis,we found that the disease location syndrome element "liver" has the greatest connection with other syndrome elements,and can form syndromes with qi deficiency,blood stasis,dampness,spleen,heat,qi stagnation,yin deficiency,kidney,gallbladder,and collaterals:liver depression spleen deficiency syndrome,liver kidney yin deficiency syndrome There are a total of 12 syndrome combinations including liver and gallbladder dampness heat syndrome,liver heat blood stasis syndrome,liver stasis spleen deficiency syndrome,liver stasis qi stagnation syndrome,liver blood stasis obstruction syndrome,liver yin deficiency syndrome,liver spleen deficiency syndrome,liver stasis spleen deficiency syndrome,liver stagnation spleen deficiency stasis blood stasis obstruction collaterals syndrome.3.374 primary liver cancer samples and 50 matched paracancerous samples were downloaded from the DCG database,and they were sorted out to distinguish between mRNA and LncRNA.Through correlation test,we screened LncRNA related to endoplasmic reticulum stress,and a total of 220 LncRNAs were screened.At the same time,we also downloaded tumor mutation data and clinical survival data related to primary liver cancer,and differentiated high and low risk groups for joint analysis,The analysis results showed that the survival rate of patients in the low-risk group was much higher than that in the high-risk group in terms of survival period,which also proved the effectiveness of the established LncRNA prognosis model for endoplasmic reticulum stress.In order to further explore the impact of endoplasmic reticulum stress related LncRNA on primary liver cancer,we conducted GO,Kegg,GSEA enrichment analysis on genes involved in the sample,and found that they were mainly related to the immune system.Tumor microenvironment analysis,immune cell correlation analysis,and immune checkpoint analysis showed that endoplasmic reticulum stress related LncRNA was mainly related to apoptosis,intracellular regulation,and immune cell activation,And this part of the direction has also been a hot topic in tumor research in recent years.4.The differential genes of LncRNA related to stress in the endoplasmic reticulum of primary liver cancer obtained in the first section of Chapter 3,as well as the related genes obtained from Gencards,OMIM,Dis Ge Net.After combining the data and deleting the duplicate values,we finally obtained 570 related genes.Through protein egg interaction network analysis of the above genes,we obtained a total of 220 effective connection targets,and used Cytoscape to reconstruct the target protein network,Obtain 13 potential core targets:PIK3CA,IL2,GF,TP53,HRAS,NRAS,KRAS,EGFR,MAPK1,BRCA1,PTEN,CTNNB1,SFN,and conduct GO and KEGG enrichment analysis on the 220 targets obtained.GO analysis results show that in biological processes,the top 5 enriched functions are:glandular development,positive regulation of DNA metabolism,positive regulation of cell adhesion,regulation of cell cycle phase transition,and intrinsic apoptosis signaling pathway;The top 5 pathways of molecular function are:transmembrane receptor protein tyrosine kinase activity,transmembrane receptor protein kinase activity,catalytic activity,acting on DNA,DNA binding transcription factor binding,mismatch repair complex binding;The top 5 pathways in cell composition analysis are:chromosomal region,DNA repair complex,thin membrane raft,membrane microregion,chromosome,telomere region.KEGG enrichment analysis found that the top 20 pathways are mostly tumor related,including multiple tumor diseases such as primary liver cancer and anti-tumor drug resistance,such as EGFR tyrosine kinase inhibitor resistance,platinum resistance,and central carbon metabolism in cancer Based on 29 genes involved in the primary liver cancer pathway,including proteoglycans and cellular aging,and related pathways related to tumor occurrence,development,and drug resistance in cancer,ITCM was used to mine traditional Chinese medicine and components.133 effective chemical components were obtained,involving 502 traditional Chinese medicine.After network graph analysis,it was found that the top 6 core genes were TP53,MAPK1,CCND1,AKT1,EGFR,and CTNNB1,respectively.The top chemical components are August acid,17 beta estradiol,michelenolide,aspirin,Capsaicin,phorbol-12-benzoate-l3-benzoate.After molecular docking,it was found that the Affinity values of 33 groups of receptor ligand docking results were all less than-5kcal-mol-1.Through molecular dynamics simulation,We observed that six groups of kcal ·mol-1<-9 receptor ligand were performing 100000 steps of isothermal isovolumic ensemble(NVT)equilibrium and Isothermal-isobaric ensemble(NPT)equilibrium,with a coupling constant of 0.1 ps and a duration of 100 ps.Finally,a free molecular dynamics simulation was run,which involved a total of 5000000 steps with a step size of 2 fs and a total timing of 100 ns.The process maintained structural stability without any ligand detachment from the receptor,demonstrating good connectivity between the receptor ligand.Conclusion1.In this study,we used Mendelian randomized research method to analyze the exposure factors related to the "improper diet" and "anger hurts the liver" of primary liver cancer,and finally found that the two exposure factors of low density lipoprotein cholesterol and liver transaminase had a direct causal relationship with primary liver cancer,which means that when the two factors involved in eating fat,sweet and thick taste.carrying out or eating by mistake affect the liver transaminase drugs or food,It can directly affect the incidence of primary liver cancer,thereby proving the direct causal relationship between the traditional Chinese medicine etiology of "improper diet" and primary liver cancer.2.By classifying and analyzing the TCM syndromes and elements of primary liver cancer,the common TCM syndromes and combinations of common syndromes were summarized,providing reference opinions for the standardization of TCM syndromes of primary liver cancer.3.The effect of endoplasmic reticulum stress lncRNA on primary liver cancer is multifaceted,which has both positive and negative regulation relationships.At the same time,it has an important impact on the survival rate of patients with primary liver cancer.The in-depth study of endoplasmic reticulum stress lncRNA may provide new ideas for the immunotherapy of primary liver cancer,inducing tumor cell apoptosis,and reducing anti tumor drug resistance.4.Based on computer-aided drug design methods,we obtained 6 core compounds and more than 20 traditional Chinese medicines,and conducted static molecular docking simulation and dynamic molecular dynamics simulation experiments.The experiments proved that under the set position and temperature conditions,the protein and compound can stably bind without ligand detachment from the protein,demonstrating its potential as a therapeutic drug for primary liver cancer.Through literature review of the six traditional Chinese medicines,we found that they all have anti-tumor effects in pharmacology,and most of them are related to liver tumors.