Title: Experimental Study on Apoptosis Induced by Survivin-antisense Oligonucleotide in Colon Cancer Cell Lines
Abstract: Survivin is a member of inhibitor of apoptosis protein family, which is highly expressed in cancerous tissues and during human embryonic and fetal development, but absent or very lowly expressed in all the mature tissues. Study had shown that survivin gene was expressed in 64.5%~85% of colonic cancerous tissues, and the expression of survivin was associated with unfavorable prognosis in colon cancer patients. Aims: To observe the effects of survivin-antisense oligonucleotide on the apoptosis of colon cancer cell lines. Methods: Through genetic engineering techniques, the survivin-antisense oligonucleotide plasmid pcDNA3-sur-As was constructed, then the plasmid DNA was transfected into colon cancer cell lines by Lipofectamine 2000. The survivin mRNA and protein expression levels in the colon cancer cell lines were determined by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot, respectively. Flow cytometry and propidium iodide (PI) staining were used to detect cell apoptosis. The variety of caspase-3 activity was determined by colorimetric assay. Results: Expression of survivin mRNA and protein were detected in the four tested colon cancer cell lines including SW1116, COLO 205, HT-29 and SW1417. After transient transfection of survivin-antisense oligo-nucleotide, the cell apoptosis rate was obviously increased in all colon cancer cell lines in a dose-dependent manner (P0.01); Typical characteristics of apoptotic cells was shown in colon cancer cell line SW1116 under fluorescence microscope. The expression level of survivin protein in SW1116 cells was decreased, and the activity of caspase-3 was significantly increased (P0.01). Conclusions: Inhibition of survivin gene expression by survivin-antisense oligonucleotide may induce apoptosis in colon cancer cell lines. Survivin targeted therapeutic protocol may possibly benefit the treatment of patients with colon cancer.
Publication Year: 2004
Publication Date: 2004-01-01
Language: en
Type: article
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