Preliminary Study On The Relationship Between Serum Small Molecule Metabolites And Lymph Node Metastasis Of Colorectal Carcinoma

Objective:Differential metabolites are investigated in colorectal cancer patients with and without lymph node metastasis to explore the ability of metabolites to differentiate colorectal cancer in different lymph node metastases.We finally expect to explore the biological mechanism of small molecule metabolites in the prediction of lymph node metastasis of colorectal cancer,and to provide a theoretical basis for predicting the lymph node metastasis of colorectal cancer.The establishment of colorectal cancer with lymph node metastasis predictive logistic regression model based on metabolites and clinical data.Method:1.LC-MS analysis and data analysis:The supernatant was detected by chromatographic mass spectrometry to obtain the expression abundance of metabolites in patients with different lymph node staging.Statistical analysis was performed by principal component analysis,t test,cluster analysis and other methods to find out the differentiated metabolites.And then re-group the samples according to the differentiated metabolites.2.Immunohistochemistry(IHC):The expression of VEGF-C and VEGF-D and the lymphatic vessel density(LVD)was detected by immunohistochemical staining.Statistical analysis was performed using the statistical software SPSS17.0.3.Enzyme-linked immunosorbent assay(ELISA):The levels of VEGF-C and VEGF-D in serum were measured by enzyme-linked immunosorbent assay(ELISA).Statistical analysis was performed using the statistical software SPSS17.0.Results:1.The metabolites of serum in patients with T3N0 and T3N1-3colorectal cancer were measured by high performance liquid chromatography-mass spectrometry.Six kinds of different metabolites were found by statistical analysis.These metabolites can well distinguish the different status of lymph node metastasis in colorectal cancer.2.Based on six different metabolites,56 samples of T3N0-3 patients with colorectal cancer were randomly divided into A and B groups.The positive samples of lymph node metastasis in group B were significantly higher than those in group A(P = 0.000),VEGF-C(P=0.007)、D(P =0.014)and lymphatic vessel density(P = 0.001)in group B.There was no significant difference in the levels of VEGF-C(P = 0.339)and VEGF-D(P = 0.451)in serum of group A and B.The overall survival rate of group A was significantly higher than that of group B(P = 0.04).3.Six kinds of independent metabolites and clinical pathologic data were used to select the six independent risk factors of lymph node metastasis of colorectal cancer by multivariate logistic regression.The risk score model was established according to the independent risk factors.The model had a good predictive ability for lymph node metastasis of colorectal cancer.Conclusion:1.Differential metabolites exist in serum levels of patients with colorectal cancer with different lymph node metastases status.2.Metabolites can well distinguish the patients of colorectal cancer with and without lymph node metastasis.3.The expression of VEGF-C and D in colorectal cancer lymph node metastasis was correlated with the expression of VEGF-C and D in the adjacent tissues,but not with the concentration of VEGF-C and D in serum.4.The risk score model which established by the independent risk factors according to the clinical pathology data and metabolites had a good predictive ability for lymph node metastasis of colorectal cancer.