[Survivin antisense oligonucleotide induces lymphoma cells apoptosis and sensitizes the cells to chemotherapy].
Hongling Li, Bing-zhong Sun, Hui Wang
Abstract
Hongling Li, Bing-zhong Sun, Hui Wang
Abstract
OBJECTIVE: To explore the effect of antisense oligodeoxynucleotide (ASODN) of survivin gene on apoptosis and chemotherapy sensitivity of lymphoma cell line Raji. METHODS: Anti-survivin phosphorothioate ASODN was synthesized and transfected into Raji cells by lipofectin. MTT assay was used to detect cytotoxicity. Apoptosis was observed by fluorescence microscopy and flow cytometry. Survivin expression was determined by RT-PCR and Western-blotting. RESULTS: (1) survivin ASODN inhibited the cells proliferation in a dose and time dependent manner. (2) A higher apoptosis rate (33.0%) could be induced in Raji cells by survivin ASODN as compared with that induced by the sense oligodeoxynucleotide (11.5%) (P < 0.05). (3) The expression of survivin mRNA and protein significantly decreased after treatment with survivin ASODN. (4) There was a significant increase of cell inhibition rate after exposure to the combination of survivin ASODN and Vm26 as compared to Vm26 or survivin ASODN alone (both P < 0.05). CONCLUSION: Survivin ASODN is able to inhibit the proliferation of Raji cells, induce the apoptosis, and enhance the sensitivity of Raji cell to chemotherapy via specific down-regulation of survivin expression.
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OBJECTIVE: To explore the effect of antisense oligodeoxynucleotide (ASODN) of survivin gene on apoptosis and chemotherapy sensitivity of lymphoma cell line Raji. METHODS: Anti-survivin phosphorothioate ASODN was synthesized and transfected into Raji cells by lipofectin. MTT assay was used to detect cytotoxicity. Apoptosis was observed by fluorescence microscopy and flow cytometry. Survivin expression was determined by RT-PCR and Western-blotting. RESULTS: (1) survivin ASODN inhibited the cells proliferation in a dose and time dependent manner. (2) A higher apoptosis rate (33.0%) could be induced in Raji cells by survivin ASODN as compared with that induced by the sense oligodeoxynucleotide (11.5%) (P < 0.05). (3) The expression of survivin mRNA and protein significantly decreased after treatment with survivin ASODN. (4) There was a significant increase of cell inhibition rate after exposure to the combination of survivin ASODN and Vm26 as compared to Vm26 or survivin ASODN alone (both P < 0.05). CONCLUSION: Survivin ASODN is able to inhibit the proliferation of Raji cells, induce the apoptosis, and enhance the sensitivity of Raji cell to chemotherapy via specific down-regulation of survivin expression.
Key concepts: Survivin, Raji cell, Apoptosis, Molecular biology, Transfection, Chemistry, Flow cytometry, Cancer research