survivin反义寡核苷酸诱导淋巴瘤细胞凋亡及对化疗药的增敏作用

Survivin antisense oligonucleotide induces lymphoma cells apoptosis and sensitizes the cells to chemotherapy

目的 观察survivin反义寡核苷酸 (ASODN)对淋巴瘤细胞的增殖、凋亡及对化疗药物敏感性的影响。方法 人工合成survivin硫代ASODN ,通过脂质体转染淋巴瘤细胞株Raji后 ,用MTT法检测细胞毒作用 ;用荧光染色、流式细胞术检测细胞凋亡 ;RT PCR、Westernblot检测survivinmRNA及蛋白的表达。结果 ①survivinASODN抑制Raji细胞增殖呈时间和剂量依赖性 ;②survivinASODN处理组Raji细胞凋亡率为 33.0 % ,正义寡核苷酸 (SODN)组为 11.5 % ,两组相比 ,差异有显著性 (P <0 0 5 ) ;③survivinASODN处理组Raji细胞survivin蛋白及mRNA的表达水平显著下降 ;④survivinASODN和Vm2 6联合处理组Raji细胞增殖受抑率为 5 6 .5 % ,显著高于单用survivinASODN组的 2 9.2 % (P <0 0 5 )和单用Vm2 6处理组的 2 6 .9% (P <0 .0 5 )。结论 survivinASODN能够抑制Raji细胞增殖、诱导凋亡 ,并能增加Raji细胞对化疗药物的敏感性 ,推测可能通过下调survivinmRNA及蛋白表达而起作用。

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 ①survivin ASODN inhibited the cells proliferation in a dose and time dependent manner. ②A 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). ③The expression of survivin mRNA and protein significantly decreased after treatment with survivin ASODN. ④There 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.