Observation On TCM Syndromes Of Colorectal Cancer And Proteomics Analysis Of Spleen Deficiency Tissue With ITRAQ

Objective The first part: 76 cases of colorectal cancer(CRC)were investigated in the Department of Gastrointestinal Surgery and Tumor Surgery,Affiliated Hospital of Armed Police Logistics College from April 2015 to September 2016.Exploring the general condition,histopathological type,the common pathological types and TCM syndromes of patients with colorectal cancer,which will lay the foundation for further study of syndrome-related proteomics.Part 2: To study the differentially expressed proteins in the tissues of patients with spleen deficiency and damp-heat type colorectal cancer,to find the objective basis of spleen deficiency and damp-heat type colorectal cancer,and to lay the foundation for further early diagnosis.In the third part,a group of markers with diagnostic and / or prognostic value can be used as therapeutic targets to improve the expression of the protein by Western Blot after the bioinformatics analysis of the tissue protein markers.In order to improve the early diagnosis and prognosis of patients or treatment possible.Methods1.Designing questionnaire,Through collecting and filling out the four patients with colorectal cancer data and the four diagnostic data dialectical to determine the TCM syndrome;2.We selected 23 patients with colorectal cancer diagnosed by biopsy and the 23 cases with spleen dampness-dampness type according to the TCM syndrome type of colorectal cancer.The i TRAQ quantitative protein analysis,and the differential proteins were analyzed by Uniprot database.The differential proteins were analyzed by bioinformatics.3.Western Blot analysis was performed on bioinformatic analysis of protein markers.Results First part:Collected 76 cases of colorectal cancer patients,more men than women,mainly in the elderly,the course of them was less than 1 month,Moderately differentiated adenocarcinoma is more common,TNM staging in stage Ⅲ,colorectal cancer patients symptom analysis was Out: the main symptoms of abdominal pain,abdominal distension,loose stools,then increased,difficulty in defecation with melena,mucus pus and blood,tenesmus,eat less,hunger does not want to eat,limb fatigue fatigue-based.Common Tongue sort tongue pink>tongue dark> tongue pale> red tongue> tongue stasis points;common tongue coating greasy moss greasy> moss thin white> moss greasy> moss slippery;common tongue sorting tooth tongue> Pulse slippery> pulse slippery> slippery pulse> slippery pulse> astringent pulse> slippery pulse> slippery pulse> pulse slippery number,the whole is slippery pulse and sinking fine vein more common.The spleen deficiency type of TCM syndromes are more common.The second part:1.The results of differential protein identification in patients with colorectal cancer The quantitative analysis of i TRAQ protein in 23 patients with spleen deficiency and dampness colorectal cancer by LC-MS / MS method showed that2771 proteins were selected.Expressing the difference Fold higher than 1.2 fold(up-down)and P-value less than 0.05 selection criteria of the protein as a difference protein,obtained 130 up,141 down.2.Screening the differentially expressed proteins of the spleen deficiency type of patients with colorectal cancer Bioinformatics analysis(including GO analysis,KEGG pathway analysis),using Fisher exact test(Fisher’s Exact Test)to conduct a significant enrichment analysis shows that: participation in the biological process,analysis,cell components mainly involved in inflammation,Immune response,cell adhesion,proliferation,apoptosis and so on.Also,Fisher’s Exact Test was used to enrich the KEGG pathway,which included 16 pathways including Focal adhesion,Spliceosome pathway and Vascular smooth muscle contraction.In addition,we performed protein quantification and differential analysis of proteins involved in the pathway.Three proteins were up-regulated in glutathione metabolism(MGST2,RRM2,SRM),four proteins were downregulated(GSTT1,GPX3,GSTM3,SMS).The third part:The results of western blot showed that RRM2,SRM,GSTM3 were consistent with the pre-i TRAQ results,and the results of SPSS19.0 software Statistical analysis found that the difference was statistically significant.Conclusions1.Without surgical treatment of colorectal cancer patients with TCM type distribution of spleen deficiency damp type more common.2.The differential expressed protein profiles between the tissues of the patients with spleen deficiency and dampness-type colorectal cancer and the normal group were described.The results showed that RRM2 and SRM were up-regulated in cancer tissues,GSTM3 was down-regulated in cancer tissues,which provides a new clue for the development and development of the spleen dampness-type colorectal cancer and the related tumor markers,and also providing early diagnosis and prognosis of great value to the colorectal cancerpatients.3.The difference proteins between the colorectal cancer patients with spleen deficiency syndrome and the normal control may be the external evidence of the intrinsic material basis.