AIM: To study the effects of sense and antisense KAI1 genes on the growth and invasion of human hepatocellular carcinoma (HCC) cell line MHCC97-H.METHODS: KAI1 sense and antisense eukaryotic expression plasmids were c...AIM: To study the effects of sense and antisense KAI1 genes on the growth and invasion of human hepatocellular carcinoma (HCC) cell line MHCC97-H.METHODS: KAI1 sense and antisense eukaryotic expression plasmids were constructed using subclone technique and transfected into MHCC97-H cells respectively by DOTAP liposome. After successful transfection was confirmed, in vitro growth curve, cell cycles, plate clone formation efficiency, invasive ability in Boyden Chamber assay and ultrastructural morphology were studied.RESULTS: KAI1 sense and antisense genes had no significant effects on the cell growth curve and cell cycles.After transfection with sense KAI1 gene, decreased invasive ability in Boyden Chamber assay and decreased amount of mitochondria, but no significant changes of plate clone formation efficiency were observed in MHCC97-H-S cells.The plate clone formation efficiency and invasive ability in Boyden Chamber assay were significantly increased in MHCC97-H-AS cells, after transfection with antisense KAI1 gene. Furthermore, increased amount of mitochondria,rough endoplasmic reticulum, Golgi apparatus and expanded endoplasmic reticulum were also noted in MHCC97-H-AS cells.CONCLUSION: Changes of KAI1 expression in HCC cells may alter their invasive and metastasis ability of the tumor.展开更多
Objective: To assess the anti-invasive effect of DDB and its possible active mechanism in human hepatocellular carcinoma MHCC97-H with high metastasis potential. Methods: MTT assay was used to evaluate the cytotoxicit...Objective: To assess the anti-invasive effect of DDB and its possible active mechanism in human hepatocellular carcinoma MHCC97-H with high metastasis potential. Methods: MTT assay was used to evaluate the cytotoxicity of DDB to MHCC97-H cells and the anti-adhesion of DDB on MHCC97-H cells to laminin (LN) and fibronectin (FN). The anti-invasive effect of DDB was detected by the transwell chamber experiment. VEGF, nm23-H1 and uPAR mRNA transcriptions were determined by RT-PCR assay. The secretion and expression of α-fetal protein (AFP) were analyzed by ELISA and flow cytometry, respectively. Results: DDB at non-cytotoxic concentrations (10, 50 and 100 μmol/L) obviously inhibited the adhe- sion of MHCC97-H on LN and FN. In the transwell chamber experiment, the inhibition rates of the invasion of DDB 50 and 100 μmol/L on MHCC97-H cells were 25.8% and 32.3%, respectively. By RT-PCR assay, DDB 50 and 100 μmol/L decreased VEGF, nm23-H1 and uPAR mRNA expressions in MHCC97-H cells. The ELISA assay showed that 50, 100 and 200 μmol/L DDB decreased the AFP secretion of MHCC97-H cells, the inhibitory rates were 16.5%, 17.5% and 48.5%, respectively. DDB also decreased the expression of AFP in MHCC97-H cells by flow cytometry assay. Conclusion: DDB, an anti-hepatitis drug, at non-cytotoxic concentrations showed significant anti-invasion effect in human hepatocellular carcinoma MHCC97-H cells, and the inhibition of VEGF, nm23-H1 and uPAR expression should contribute to the anti-invasion property of DDB.展开更多
基金Supported by the National Natural Science Foundation of China,No.30070348
文摘AIM: To study the effects of sense and antisense KAI1 genes on the growth and invasion of human hepatocellular carcinoma (HCC) cell line MHCC97-H.METHODS: KAI1 sense and antisense eukaryotic expression plasmids were constructed using subclone technique and transfected into MHCC97-H cells respectively by DOTAP liposome. After successful transfection was confirmed, in vitro growth curve, cell cycles, plate clone formation efficiency, invasive ability in Boyden Chamber assay and ultrastructural morphology were studied.RESULTS: KAI1 sense and antisense genes had no significant effects on the cell growth curve and cell cycles.After transfection with sense KAI1 gene, decreased invasive ability in Boyden Chamber assay and decreased amount of mitochondria, but no significant changes of plate clone formation efficiency were observed in MHCC97-H-S cells.The plate clone formation efficiency and invasive ability in Boyden Chamber assay were significantly increased in MHCC97-H-AS cells, after transfection with antisense KAI1 gene. Furthermore, increased amount of mitochondria,rough endoplasmic reticulum, Golgi apparatus and expanded endoplasmic reticulum were also noted in MHCC97-H-AS cells.CONCLUSION: Changes of KAI1 expression in HCC cells may alter their invasive and metastasis ability of the tumor.
文摘Objective: To assess the anti-invasive effect of DDB and its possible active mechanism in human hepatocellular carcinoma MHCC97-H with high metastasis potential. Methods: MTT assay was used to evaluate the cytotoxicity of DDB to MHCC97-H cells and the anti-adhesion of DDB on MHCC97-H cells to laminin (LN) and fibronectin (FN). The anti-invasive effect of DDB was detected by the transwell chamber experiment. VEGF, nm23-H1 and uPAR mRNA transcriptions were determined by RT-PCR assay. The secretion and expression of α-fetal protein (AFP) were analyzed by ELISA and flow cytometry, respectively. Results: DDB at non-cytotoxic concentrations (10, 50 and 100 μmol/L) obviously inhibited the adhe- sion of MHCC97-H on LN and FN. In the transwell chamber experiment, the inhibition rates of the invasion of DDB 50 and 100 μmol/L on MHCC97-H cells were 25.8% and 32.3%, respectively. By RT-PCR assay, DDB 50 and 100 μmol/L decreased VEGF, nm23-H1 and uPAR mRNA expressions in MHCC97-H cells. The ELISA assay showed that 50, 100 and 200 μmol/L DDB decreased the AFP secretion of MHCC97-H cells, the inhibitory rates were 16.5%, 17.5% and 48.5%, respectively. DDB also decreased the expression of AFP in MHCC97-H cells by flow cytometry assay. Conclusion: DDB, an anti-hepatitis drug, at non-cytotoxic concentrations showed significant anti-invasion effect in human hepatocellular carcinoma MHCC97-H cells, and the inhibition of VEGF, nm23-H1 and uPAR expression should contribute to the anti-invasion property of DDB.
