Expression And Significance Of HnRNP AB In Colorectal Cancer

Objective:1.To investigate the expression of hnRNP AB in colorectal cancer tissues and and its relationship with clinicopathological characteristics and prognosis.2.To investigate the expression of hnRNP AB in colorectal cancer stem cells.Methods:1.To detect the expression of hnRNP AB in colorectal cancer tissues and its adjacent normal tissues from 30 colorectal cancer patients by Immunohistochemistry.2.The expression of hnRNP AB in 184 cases of colorectal cancer was detected by Immunohistochemical,and its relationship with clinicopathological features and prognosis was analyzed:(1)The relationship of hnRNP AB protein in colorectal cancer tissues and clinicopathological features including CEA,CA19-9,tumor size,depth of tumor invasion,lymph node metastasis,distant metastasis and TNM stage;(2)The difference of 5-year survival rate of patients with colorectal cancer in different expression of hnRNP AB protein;(3)One-way Cox survival analysis the factors that affected the overall survival rate(OS)of colorectal cancer;(4)Multivariate Cox analysis the independent risk factor that affected the OS of colorectal cancer.3.CD44(+)cells and CD44(-)cells were sorted by CD44 immunomagnetic beads from colorectal cancer SW480 cells:(1)The level of CD44 expression that detected by Flow cytometry was used to identify the sorting results;(2)Colorectal cancer stem cell markers CD44 and CD133 expressions in the two groups of cells were detected by Immunofluorescence;(3)The relative expression of hnRNP AB mRNA in CD44(+)cells and CD44(-)cells was detected by RT-qPCR;(4)The expression of hnRNP AB protein inCD44(+)cells and CD44(-)cells was detected by Flow cytometry?Western blot and Immunofluorescence.Results:1.Immunohistochemical results of 30 cases of colorectal cancer tissues and corresponding adjacent tissues suggest that the expression level of hnRNP AB protein(IOD/ARE value)is 0.0438±0.0044 and 0.0072±0.0017,respectively.The expression of hnRNP AB protein in colorectal cancer tissues was significantly higher than that of adjacent normal tissues(P<0.01).2.The relationship between the expression of hnRNP AB and clinicopathological features in 184 patients with colorectal cancer was analyzed:(1)The expression of hnRNP AB protein in colorectal cancer tissues were related with CEA,CA19-9,tumor size,tumor infiltration,lymph node metastasis,distant metastasis,TNM stage(P<0.05);(2)Kaplan-Meier survival curve analysis showed that the 5-year survival rate of patients with high expression of hnRNP AB in colorectal cancer was significantly lower than that of patients with low expression of hnRNP AB(P<0.01);(3)Univariate Cox survival analysis showed that factors affecting OS of colorectal cancer were CEA,CA19-9,tumor size,tumor differentiation,tumor infiltration,lymph node metastasis,distant metastasis,TNM stage and hnRNP AB(P<0.05).(4)Multivariate Cox analysis showed that high expression of hnRNP AB was an independent prognostic indicator for poor prognosis of colorectal cancer.3.CD44(+)cells and CD44(-)cells were sorted from Colorectal cancer SW480 cells by CD44 immunomagnetic beads:(1)The percentage of CD44 in CD44(+)cells was up to99% and the percentage in CD44(-)cells was only 5.6% detected by Flow cytometry,indicating that the sorting was successful;(2)Immunofluorescence assay showed that the levels of CD44 and CD133 protein in CD44(+)cells were higher than that in CD44(-)cells,it is speculated that CD44(+)cells and CD44(-)cells are colorectal cancer stem cells and colorectal cancer non-tumor stem cells,respectively;(3)The results of RT-qPCR showedthat the level of hnRNP AB mRNA in CD44(+)cells was higher than that in CD44(-)cells(P<0.05).(4)The results of Flow cytometry?Western blot and Immunofluorescence assay showed that the level of hnRNPAB protein in CD44(+)cells was higher than that of CD44(-)cells(P<0.05).Conclusion:1.The expression of hnRNP AB in colorectal cancer tissues is significantly higher than its corresponding adjacent tissues,suggesting that it is associated with the occurrence of colorectal cancer;2.The expression of hnRNP AB in colorectal cancer is related to CEA,CA19-9,tumor size,tumor infiltration,lymph node metastasis,distant metastasis and TNM stage,and it is considered as an independent risk factor affecting the prognosis of colorectal cancer,suggesting that it is associated with the progression of colorectal cancer;3.hnRNP AB is highly expressed in colorectal cancer stem cells,suggesting that it may be involved in the regulation of colorectal cancer stem cells.