Research Of The Role And Mechanism Of CD151 On The Tumor Xenograft Of Nude Mice Caused Colorectal Cancer Stem Cells

Objective: Colon cancer stem cells was used to establish the tumor xenograft of nude mice model in this study. At the cellular level and the whole animal level, It is to explore into the effects of CD151 gene on Wnt signal transduction pathway of the tumor xenograft of nude mice model.Method:1 Four kinds of cells were cultured by using cell culture technique,which is HT29 cells,CD151-HT29 cells,CD133+CD44+ colon cancer stem cells and CD151-CD133+CD44+ colon cancer stem cells.To observe the growth of cells, normal colon cancer cells and colon cancer stem cells are obtained in logarithmic growth phase dry;through re-inoculated into serum supplemented medium,colon cancer stem cells were induced to differentiation,which can identify colon cancer stem cell.2 CD44 gray values and CD133 gray values from HT29 Cells and CD151- HT29 Cells was detected by laser confocal microscopy.3 The construction method of transplantation tumor model in nude mice:Four kinds of cells were inoculated subcutaneously in nude mice.Which were HT29 cells,CD151-HT29 cells,CD133+CD44+colon cancer stem cells and CD151- CD133+ CD44+ colon cancer stem cells. the tumor growth and pathological examination were observed.4 The expression levels of Wnt-5a m RNA and beta-catenin m RNA in transplantation tumor was detected by real-time fluorescent quantitative RT-PCR technique, Transplantation tumor were caused by CD44+ CD133+group and CD151-CD44+CD133+group.Results:1 HT29 cells and CD151-HT29 cells,CD151-CD133+CD44+colon cancer stem cells and CD133+CD44+colon cancer stem cells are cultured in good growth Status.Colon cancer stem cells was re-inoculated into serum supplemented medium, cell adherence and differentiation were resumed again.2 CD44 gray values and CD133 gray values From HT29 Cells were significantly higher than CD151- HT29 Cells,which were detected by laser confocal microscopy.3 The tumor formation rate, tumor volume, degree of malignancy of CD133+ CD44+ colon cancer stem cell group were higher than that of the HT29 cells group in nude mice.4 The m RNA expression levels of β-catenin and Wnt-5a in CD44+ CD133+ group were significantly higher than CD151-CD44+CD133+group in the tumor xenograft of nude mice.Conclusion:1 Colon cancer stem cells have stong proliferation and invasion ability,which are screened by CD133 antibody and CD44 antibody;the expression of CD44 protein and CD133 protein in the knockout the CD151 gene of HT29 cells is decreased significantly compared with the original strain of HT29 cells.2 The transcription levels of Wnt-5a and beta-catenin gene are significantly decreased in the knockout CD151 gene of colon cancer stem cells groups,which are related to the regulation of the cell proliferation and differentiation, and the tumor formation rate,tumor volume, tissue malignant degree and the invasion of surrounding tissue also decreased, suggesting that CD151 gene may affect colon cancer proliferation and invasion function through Wnt pathways.3 The illumation of physiological function about CD151 and monitoring of the expression level about CD151 gene, may provide a strong basis for the future treatment of colon cancer.