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Effects of Citrus Reticulata on the Cell Detachment and Apoptosis in Human Gastric Cancer SNU-668 Cells

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The purpose of this study was to examine the effects of Citrus Reticulata(CR) on the Cell Detachment and Apoptosis in Human Gastric Cancer SNU-668 Cells. The effect of CR on apoptosis was investigated through MTT assay, DAPI staining, and TUNEL assay. We also performed RT-PCR for apoptotic genes including BCL-2, BAX, and caspase-3, the caspase-3 activity assay, and western blotting for pro-CASP-3. Then, to detect that adhesion of cell to ECM was reduced by CR, we investigated mRNA expression of CDH1 and PTK2 using RT-PCR, and their protein expressions using western blotting, and immunocytochemistry in SNU-668 cells. In this study, the results showed that treatment of CR induced time and dose-dependent cell death in SNU-668 cells. Downregulated mRNA expression of BCL-2, and upregulated mRNA expressions of BAX and CASP-3 indicated that the cell death was due to apoptosis. Protein expression of inactivated CASP-3, and caspase-3 activity assay also showed that apoptosis was induced in CR-treated cells.

Abstract

The purpose of this study was to examine the effects of Citrus Reticulata(CR) on the Cell Detachment and Apoptosis in Human Gastric Cancer SNU-668 Cells. The effect of CR on apoptosis was investigated through MTT assay, DAPI staining, and TUNEL assay. We also performed RT-PCR for apoptotic genes including BCL-2, BAX, and caspase-3, the caspase-3 activity assay, and western blotting for pro-CASP-3. Then, to detect that adhesion of cell to ECM was reduced by CR, we investigated mRNA expression of CDH1 and PTK2 using RT-PCR, and their protein expressions using western blotting, and immunocytochemistry in SNU-668 cells. In this study, the results showed that treatment of CR induced time and dose-dependent cell death in SNU-668 cells. Downregulated mRNA expression of BCL-2, and upregulated mRNA expressions of BAX and CASP-3 indicated that the cell death was due to apoptosis. Protein expression of inactivated CASP-3, and caspase-3 activity assay also showed that apoptosis was induced in CR-treated cells.

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