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目的 观察HA82 4 ,这种新合成的靶向HER 2mRNA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HA82 4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HA82 4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) :HA82 4 ,(47± 2 6 ) ;HA4 ,(97± 4 1) ;随机序列 ,(2 5 1± 86 )。HA82 4vsHA4 ,P <0 .0 5〕。HA82 4还显著抑制了HER 2mRNA与蛋白的表达 (免疫组化结果 :HA82 4 ,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 :HA82 4 ,338.6 ;HA4 ,35 5 .5 ;随机序列 ,5 32 .4 )。结论HA82 4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
Objective To investigate the specific effect of HA82 4, a newly synthesized antisense oligonucleotide targeting HER 2 mRNA, on the mRNA and protein expression of SK BR 3 breast cancer cell lines. Methods HA4 was used as a positive control and the random sequence was used as a randomized control to detect the inhibitory effect and IC50 value of HA82 4 on SK BR 3 cell growth. RT-PCR, flow cytometry, immunohistochemistry The effect on mRNA and protein expression of SK BR 3 cells was examined. The results showed that HA82 4 had a dose-dependent inhibitory effect on the growth of SK BR 3 breast cancer cells [IC50 (nmol·L-1): HA82 4, (47 ± 2 6), HA4, (97 ± 4 1) Sequence, (2 5 1 ± 86). HA82 4vsHA4, P <0.05). HA82 4 also significantly inhibited HER 2 mRNA and protein expression (immunohistochemistry results: HA82 4, 1 + staining; HA4, 2 + staining; random sequence, 3 + staining; flow cytometry results, the fluorescence intensity were: HA82 4, 338.6; HA4, 35 5 .5; random sequence, 5 32 .4). Conclusion The inhibitory effect of HA82 4 on the growth of SK BR 3 breast cancer cells is related to the specific inhibition of HER 2 expression