AIM: To investigate the effects of KAI1/CD82 on biological behavior of colorectal carcinoma cells.
METHODS: KAI1 cDNA was transfected into highly malignant colorectal carcinoma cell line, LoVo, which had low level of ...AIM: To investigate the effects of KAI1/CD82 on biological behavior of colorectal carcinoma cells.
METHODS: KAI1 cDNA was transfected into highly malignant colorectal carcinoma cell line, LoVo, which had low level of endogenous KAI1 expression, and established stable transfectant clones with high KAI1/CD82 expression.The cell-cell adhesion, cell aggregation, cell-matrix adhesion and cell invasion assay were performed to determine whether KAI1 transfectant could have an effect on proliferation,adhesion and tumor metastasis in comparison with the control transfectant cells.
RESULTS: KAI1 expression did not alter in vitro cell proliferation. But the KAI1 transfectant cells exhibited significantly increased homotypic cell-cell adhesion and cell aggregation in comparison with the control transfectant cells (P＜0.05). Furthermore, KAI1 expression significantly suppressed the cell adhesion to extracellular matrix components and in vitro cell invasion in KAI1-transfected LoVo cells. The data indicated that KAI1 expression significantly suppressed the metastatic potential of KAI1-transfected LoVo cells.
CONCLUSION: Our results suggest that KAI1 might function as a negative regulator of colorectal carcinoma metastasis.展开更多
AIM: To construct a differentially-expressed gene subtracted cDNA library from two colorectal carcinoma (CRC) cell lines with different metastatic phenotypes by suppression subtractive hybridization.METHODS: Two cell ...AIM: To construct a differentially-expressed gene subtracted cDNA library from two colorectal carcinoma (CRC) cell lines with different metastatic phenotypes by suppression subtractive hybridization.METHODS: Two cell lines of human CRC from the same patient were used. SW620 cell line showing highly metastatic potential was regarded as tester in the forward subtractive hybridization, while SW480 cell line with lowly metastatic potential was treated as tester in the reverse hybridization. Suppression subtractive hybridization (SSH) was employed to obtain cDNA fragments of differentially expressed genes for the metastasis of CRC. These fragments were ligated with T vectors, screened through the blue-white screening system to establish cDNA library.RESULTS: After the blue-white screening, 235 white clones were picked out from the positive-going hybridization and 232 from the reverse. PCR results showed that 200-700 bp inserts were seen in 98% and 91% clones from the forward and reverse hybridizations, respectively.CONCLUSIONS: A subtractive cDNA library of differentially expressed genes specific for metastasis of CRC can be constructed with SSH and T/A cloning techniques.展开更多
基金the National Natural Science Foundation,No. 31070423the Natural Science Foundation of Guangdong Province,No.990385,970335the Natural Science Foundation of PLA of China,No.01MA128
文摘AIM: To investigate the effects of KAI1/CD82 on biological behavior of colorectal carcinoma cells.
METHODS: KAI1 cDNA was transfected into highly malignant colorectal carcinoma cell line, LoVo, which had low level of endogenous KAI1 expression, and established stable transfectant clones with high KAI1/CD82 expression.The cell-cell adhesion, cell aggregation, cell-matrix adhesion and cell invasion assay were performed to determine whether KAI1 transfectant could have an effect on proliferation,adhesion and tumor metastasis in comparison with the control transfectant cells.
RESULTS: KAI1 expression did not alter in vitro cell proliferation. But the KAI1 transfectant cells exhibited significantly increased homotypic cell-cell adhesion and cell aggregation in comparison with the control transfectant cells (P＜0.05). Furthermore, KAI1 expression significantly suppressed the cell adhesion to extracellular matrix components and in vitro cell invasion in KAI1-transfected LoVo cells. The data indicated that KAI1 expression significantly suppressed the metastatic potential of KAI1-transfected LoVo cells.
CONCLUSION: Our results suggest that KAI1 might function as a negative regulator of colorectal carcinoma metastasis.
基金Supported by the Military Medical Foundation of China,No.01MA128
文摘AIM: To construct a differentially-expressed gene subtracted cDNA library from two colorectal carcinoma (CRC) cell lines with different metastatic phenotypes by suppression subtractive hybridization.METHODS: Two cell lines of human CRC from the same patient were used. SW620 cell line showing highly metastatic potential was regarded as tester in the forward subtractive hybridization, while SW480 cell line with lowly metastatic potential was treated as tester in the reverse hybridization. Suppression subtractive hybridization (SSH) was employed to obtain cDNA fragments of differentially expressed genes for the metastasis of CRC. These fragments were ligated with T vectors, screened through the blue-white screening system to establish cDNA library.RESULTS: After the blue-white screening, 235 white clones were picked out from the positive-going hybridization and 232 from the reverse. PCR results showed that 200-700 bp inserts were seen in 98% and 91% clones from the forward and reverse hybridizations, respectively.CONCLUSIONS: A subtractive cDNA library of differentially expressed genes specific for metastasis of CRC can be constructed with SSH and T/A cloning techniques.
