Study On The Distribution Characteristics Of TCM Syndrome Types And Protein Profile Analysis In Patients With Colorectal Cancer Based On Septin9 Gene Methylation

Objective:In this study,Septin9 gene methylation(mSEPT9)was used as the research point to explore the distribution characteristics of TCM syndrome types in patients with colorectal cancer,and to preliminarily explore the correlation between mSEPT9 and TCM syndrome types.In this way,we provide a theoretical basis for the clinical treatment of colorectal cancer with negative or positive Septin9 methylation test results.On the other hand,we conducted protein spectrogram analysis on colorectal cancer and adjacent normal tissues with negative or positive Septin9 methylation test results to screen out the differential proteins and identify potential biomarkers of colorectal cancer.This will be used in conjunction with the Septin9 gene methylation test for the diagnosis of colorectal cancer.Methods:1.This study was a retrospective study.We screened patients diagnosed with colorectal cancer who attended outpatient clinics or were hospitalized between January 2019 and December 2022 based on inclusion and exclusion criteria.Subsequently,according to the syndrome differentiation and classification criteria of colorectal cancer organized by the Spleen and Stomach Diseases Branch of the Chinese Association of Traditional Chinese Medicine and the patient’s medical record information,TCM syndrome types were classified,and medical record data database was established.Finally,statistical analysis was made on the data,and the distribution of TCM syndrome types in colorectal cancer patients with negative or positive Septin9 gene methylation test results and the correlation between Septin9 gene methylation and pathological changes were obtained.2.Paraffin-embedded tissues of patients with colorectal cancer were collected for pre-experiment,and then protein profiling of cancer and adjacent normal tissues of 10 samples was performed in batches.Samples of 10 patients with colorectal cancer included were divided into four groups:colorectal cancer tissue of the Msept9-negative group,adjacent normal tissue of the Msept9-negative group,colorectal cancer tissue of the Msept9-positive group,and adjacent normal tissue of the Msept9-positive group.The differential expression ratio and P value were used as indexes to screen the significantly differentially expressed proteins among the groups as potential biomarkers Finally,the core proteins were verified by immunohistochemical experiments.Results:1.A total of 69 patients with colorectal cancer were included for TCM syndrome type analysis.Statistical analysis showed that there were no significant differences in tumor location,differentiation degree,TNM stage and lymph node metastasis between mSEPT9 negative group and positive group.In terms of the distribution of TCM syndrome types,the TCM syndrome types of patients whose Septin9 gene methylation test result was negative were mainly spleen deficiency and dampness-sheng syndrome,Yin deficiency and toxic-heat syndrome and qi and blood deficiency syndrome,while the TCM syndrome types of patients whose Septin9 gene methylation test result was positive were mainly spleen deficiency and dampness-sheng syndrome,dampness-toxic stasis syndrome and qi stagnation and blood stasis syndrome.This will provide some theoretical basis for the use of drugs in patients with colorectal cancer whose clinical symptoms are not obvious and lack of dialectical evidence.2.A total of 4589 proteins were identified by protein spectrum analysis of 10 samples of cancer and adjacent normal tissues,among which 4340 proteins could be quantified.The core differentially expressed proteins including RPL15,EPDR1 and CLNS1A were obtained by juxtaposed analysis of the significantly differentially expressed proteins among the comparison groups.Immunohistochemical results showed that the three proteins could be applied to the diagnosis of MsepT9-negative colorectal cancer patients alone or in combination,with high sensitivity and specificity.Conclusions:1.The distribution of TCM syndrome types in patients with colorectal cancer whose Septin9 gene methylation test results were negative or positive was statistically different.In clinic,medication can be properly adjusted according to the different methylation test results of Septin9 gene,so as to obtain better therapeutic effect.2.RPL15,EPDR1 and CLNS1A can be used as potential markers for supplementary diagnosis of colorectal cancer patients whose Septin9 gene methylation test results were negative,and improve the sensitivity and specificity of diagnosis of colorectal cancer.