| Objective:The discovery of cancer stem cells(CSC)has improved an understanding of tumor occurrence and development.According to stem cell(SC)theory,colorectal carcinoma may be derived from mutations in normalintestinal stem cells.So CSC can be indirectly defined by the cell of origin(stem cells or early progenitor cells).Thus,through the special markers of cancer stem cell,we can determine the location and number of cancer stem cells,and also can research its relationship to various clinicopathological features and patients' survival time in human colorectal carcinoma(CRC)patients Aldehyde dehydrogenasel(ALDH1),a detoxifying enzyme responsible for the oxidation of intracellular aldehydes,a common markers of CSCs and SCs.CD133(also called as Prominin-1 or AC 133)is a transmembrane glycoprotein of 865 amino acid,with a total molecular weight of 120 kDa.In previous studies,investigtors have used monoclonal antibodies against CD133 for the identification and isolation of a putative CSC population from some malignant tumors,such as brain,prostate,liver,pancreas,lung and colon.And the CD133+cells have demonstrated increased tumorigenic potential in transplantation.Arsenic,also named arsenic trioxide(As2O3)is a main ingredient of numerous traditional Chinese herbal recipes.It has a role in promoting tumor cell differentiation,inhibition of angiogenesis and tumor cell metastasis.Thro ugh immunohistochemistry and indirect florescence antibody staining,the aim of this experiment was done to explore the value of CSC markers-ALDH1 and CD133 in human colorectal carcinoma,also studied the role of Chinese medicine arsenic affects the tumor cells and CSC for colorectal carcinoma in primary cell cultures,which do for the aim to cure cancer and enhance the survival time of patients.Methods:We investigated the relationship between ALDH1 and CD133 protein expression with the various clinicopathological parameters through immunohistochemistry(IHC)and indirect florescence antibody staining(IFAS)in wax block organization.Furthermore,we used the Kaplan-Meier method to estimate patient's overall survival.Correlation of survival differences between ALDH1-positive or CD133-positive expression and negative control was analyzed by the K-M test.Through spearman's rank analyses study the relationship between ALDH and CD133 expression.Special researched the features of ALDH1 in rectal cancer.Meantime by primary colorectal tissue staining for cell culture,want to through the markers ALDH1 of CSC to identify CSC,furthermore tumor cell culture was added medicine arsenic(As2O3),research its impact on colorectal cancer cells and cancer stem cells.We used Enzyme-Linked Immunosorbnent Assay(ELISA)and western blot(WB)to investigate the different content of ALDH1 between fresh colorectal cancer tissues and fresh normal colorectal tissues.Results:Human colorectal carcinoma wax block tissue exhibited dramatically higher levels of ALDH1 protein compared with normal colorectal tissue.We demonstrated that a strong correlation between differentiation degree of tumor' cell and the expression of ALDH1,the lower degree of cell differentiation,the higher expression of ALDH1.But CD133 did not show the same results.We also found that low-stage tumor showed higher expression of ALDH1 or CD133 staining compared to high-stage tumors.CD133 expression was related with lymph node metastasis cases,but ALDH1 did not show the feature.Through Kaplan-Meier analysis for overall survival,we demonstrated that compared to negative cases,patients with ALDH1-positive had a poor prognosis.Arsenic added to the culture medium significantly inhibited the growth of tumor cells,prevented the CSC transform to cancer cells,as results increasing the intensity of the CSC.Moreover,as compared with fresh normal colorectal tissue,ELISA assay demonstrated there was more ALDH1 in colorectal tissue homogenates.CD 133-positive cases did not show any significance than CD133-negative cases in survival period,both were bad.The expression of ALDH1 and CD133 were significantly correlated.Conclusion:Through research,we found more ALDH1 in colorectal tissue than in normal tissue.,also found the the telationship of ALDH1 and CD133 protein expression and cl ini copatho logical features in human colorectal carcinoma,which show that ALDH1 or CD133 expression is strongly associated with tumor clinic stage and tumor cell differentiation.CD133 associate with lymph node metastasis.Patients with ALDH1-positive had a poor prognosis,but CD133-positive did not show the same results.Therefore,combined detection ALDH1 and CD133 can be served as very useful markers of CSC to identify CSC.Medicine arsenic,as a useful medicine kills a lot of cancer cells,meantime enhance the intensity of CSC for it preventting the CSC transform to cancer cell.By using the medicine arsenic(As2O3),can help us isolate or purify CSC. |
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