目的观察HA824,这种新合成的靶向HER-2 mRNA的反义寡核苷酸对SK-BR-3乳癌细胞株mRNA和蛋白表达的特异性作用.方法以HA4为阳性对照,随机序列为随机对照检测了HA824对SK-BR-3细胞生长的抑制作用与IC50值;采用逆转录聚合酶链反应(RT-PCR)、流式细胞技术、免疫组织化学法检测了对SK-BR-3细胞mRNA与蛋白表达的影响.结果结果表明HA824对SK-BR-3乳癌细胞的生长有剂量依赖性的抑制作用[IC50(nmol@L-1):HA824,(47±26);HA4,(97±41);随机序列,(251±86).HA824 vs HA4,P＜0.05].HA824还显著抑制了HER-2 mRNA与蛋白的表达(免疫组化结果:HA824,1+染色;HA4,2+染色;随机序列,3+染色;流式细胞技术检测结果,荧光强度分别为:HA824,338.6;HA4,355.5;随机序列,532.4).结论HA824对SK-BR-3乳癌细胞生长的抑制作用与特异性的抑制HER-2表达有关.%AIM To observe specific effect of HA824, a new synthesized antisense oligodeoxyribonucleotide (ODN), HA824, targeting at HER-2 mRNA on HER-2expression in SK-BR-3 breast cancer cells in mRNA and protein levels. METHODS Antisense ODN HA824 targeting at HER-2 mRNA was synthesized as our previous description. Growth inhibition effect, mean ( n = 3 - 5)50% inhibitory concentrations (IC50) on proliferations of SK-BR-3 cells for HA824, HA4 and scramble were evaluated. HA824 on HER-2 mRNA and protein expression were examined by means of RT-PCR, flow cytometry and immunohistochemistry. RESULTS The results indicated HA824 inhibited proliferation of SK-BR-3 cells in a dose dependent manner [ IC50 ( nmol@ L- 1 ): HA824, (47 ±26); HA4, (97±41); scramble, (251 ±86). HA824 vs HA4, P ＜ 0.05 ]. HA824 also significantly suppressed HER-2 mRNA and protein expression detected by RT-PCR, immunohistochemistry(HA824, light staining;HA4, moderate staining; scramble, heavy staining) and flow cytometry ( fluorescence intensity: HA824, 338.6;HA4, 355.5; scramble, 532.4). CONCLUSION Not only can this kind of new synthesized antisense compound suppressed proliferation of SK-BR-3 breast cancer cells,but also specially inhibited HER-2 expression in mRNA and protein levels.
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