[Effect of recombinant caspase-3 gene transfection on CNE2 cell apoptosis].
Xiao-xia Wang, Xiaobao Yao, Xian-sheng Ji, Hongliang Zhu
Abstract
Xiao-xia Wang, Xiaobao Yao, Xian-sheng Ji, Hongliang Zhu
Abstract
OBJECTIVE: To investigate the apoptosis-inducing effect of caspase-3 on human nasopharyngeal carcinoma cells (CNE2). METHODS: Recombinant caspase-3 was subcloned into the eukaryotic expression vector PEGFP-C1 containing the reporter gene using DNA recombinant technique. CNE2 cells were transfected with the recombinant caspase-3 gene via lipofectamine 2000 and the expression of caspase-3 mRNA was detected by reverse transcription-polymerase chain reaction. The cell morphological changes were observed under fluorescence microscope and electron microscope and the cell survival rate after the transfection was assessed by MTT assay. RESULTS: Transfection with the recombinant caspase-3 gene induced significant apoptosis in CNE2 cells, which exhibited obvious morphological changes typical of apoptotic cells. CONCLUSION: The recombinant caspase-3 gene can inhibit the growth and effectively induce apoptosis of CNE2 cells.
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OBJECTIVE: To investigate the apoptosis-inducing effect of caspase-3 on human nasopharyngeal carcinoma cells (CNE2). METHODS: Recombinant caspase-3 was subcloned into the eukaryotic expression vector PEGFP-C1 containing the reporter gene using DNA recombinant technique. CNE2 cells were transfected with the recombinant caspase-3 gene via lipofectamine 2000 and the expression of caspase-3 mRNA was detected by reverse transcription-polymerase chain reaction. The cell morphological changes were observed under fluorescence microscope and electron microscope and the cell survival rate after the transfection was assessed by MTT assay. RESULTS: Transfection with the recombinant caspase-3 gene induced significant apoptosis in CNE2 cells, which exhibited obvious morphological changes typical of apoptotic cells. CONCLUSION: The recombinant caspase-3 gene can inhibit the growth and effectively induce apoptosis of CNE2 cells.
Key concepts: Lipofectamine, Transfection, Molecular biology, Recombinant DNA, Apoptosis, Biology, MTT assay, Chemistry