2001Chieh P'ou Hsueh PaoRequires access

INHIBITION OF T-CELL APOPTOSIS BY TRANSFECTING ANTISENSE HUMAN Fas GENE AND ITS ROLE OF TUMOR THERAPY

Hong Zhang

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Abstract

Objective By inhibiting approach of apoptosis of T cells to observe if it could reduce the attacks of tumor cells upon lymphocytes and its possible role in tumor treatment. Methods The expressions of Fas/FasL genes in ovarian carcinoma cells were detected by using flow cytometry and RT\|PCR method.The construction of pcDNA3\|antisense Fas,the constructed vector was transfected into Jurkat cells with lipofectin,the change in expression of Fas gene was determined by flow cytometry.By means of Annexin\|V and MTT the effect of apoptosis in transduced Jurkat cells was investigated,and also using MTT cytotoxic test to investigate how 3AO cells kill Jurkat cells. Results Fas/FasL were expressed in 6 ovarian carcinomal cells.The expression level of Fas protein in Jurkat cells transduced with the constructed vector was decreased.Apoptosis was reduced in antisense Fas\|transfected Jurkat cells after anti\|Fas treatment. Conclusion FasL expression in ovarian carcinoma may be one of the reasons for ovarian carcinoma to escape immunosurveillance and attacking lymphocytes.Blocking Fas expression in lymphocytes can partially inhibit Jurkat cells apoptosis induced by anti\|Fas and reducing the attacks of tumor cells upon lymphocyte.\;

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Objective By inhibiting approach of apoptosis of T cells to observe if it could reduce the attacks of tumor cells upon lymphocytes and its possible role in tumor treatment. Methods The expressions of Fas/FasL genes in ovarian carcinoma cells were detected by using flow cytometry and RT\|PCR method.The construction of pcDNA3\|antisense Fas,the constructed vector was transfected into Jurkat cells with lipofectin,the change in expression of Fas gene was determined by flow cytometry.By means of Annexin\|V and MTT the effect of apoptosis in transduced Jurkat cells was investigated,and also using MTT cytotoxic test to investigate how 3AO cells kill Jurkat cells. Results Fas/FasL were expressed in 6 ovarian carcinomal cells.The expression level of Fas protein in Jurkat cells transduced with the constructed vector was decreased.Apoptosis was reduced in antisense Fas\|transfected Jurkat cells after anti\|Fas treatment. Conclusion FasL expression in ovarian carcinoma may be one of the reasons for ovarian carcinoma to escape immunosurveillance and attacking lymphocytes.Blocking Fas expression in lymphocytes can partially inhibit Jurkat cells apoptosis induced by anti\|Fas and reducing the attacks of tumor cells upon lymphocyte.\;

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Available abstract

Objective By inhibiting approach of apoptosis of T cells to observe if it could reduce the attacks of tumor cells upon lymphocytes and its possible role in tumor treatment. Methods The expressions of Fas/FasL genes in ovarian carcinoma cells were detected by using flow cytometry and RT\|PCR method.The construction of pcDNA3\|antisense Fas,the constructed vector was transfected into Jurkat cells with lipofectin,the change in expression of Fas gene was determined by flow cytometry.By means of Annexin\|V and MTT the effect of apoptosis in transduced Jurkat cells was investigated,and also using MTT cytotoxic test to investigate how 3AO cells kill Jurkat cells. Results Fas/FasL were expressed in 6 ovarian carcinomal cells.The expression level of Fas protein in Jurkat cells transduced with the constructed vector was decreased.Apoptosis was reduced in antisense Fas\|transfected Jurkat cells after anti\|Fas treatment. Conclusion FasL expression in ovarian carcinoma may be one of the reasons for ovarian carcinoma to escape immunosurveillance and attacking lymphocytes.Blocking Fas expression in lymphocytes can partially inhibit Jurkat cells apoptosis induced by anti\|Fas and reducing the attacks of tumor cells upon lymphocyte.\;

Key concepts: Jurkat cells, Fas ligand, Apoptosis, Transfection, Flow cytometry, Cytotoxic T cell, Molecular biology, Biology

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