靶向HER-2 mRNA反义寡核苷酸对SK-BR-3乳腺癌细胞Caspase-3蛋白表达的影响

目的　检测HER 2特异性的反义寡核苷酸HA82 4对SK BR 3乳腺癌细胞Caspase 3蛋白表达的影响。 方法　选用HER 2高表达的SK BR 3乳腺癌细胞株作为试验的细胞株。HA82 4合成如前所述 ,HA4为已报道的阳性序列 ,Scramble设定为随机对照序列。上述药物分别以浓度为 2 0 0nmol·L-1孵育SK BR 3细胞 8h和 36h ,之后采用逆转录PCR法与免疫细胞化学技术检测SK BR 3细胞HER 2mR NA及Caspase 3蛋白表达水平。 结果　与Scramble对照序列相比 ,HA82 4、HA4能抑制HER 2mRNA的表达 ,HA82 4作用更强 ;与Scramble相比 ,HA82 4、HA4对Caspase 3蛋白表达水平则无影响。结论　HA82 4、HA4这两种靶向HER 2mRNA反义药物对SK BR 3乳腺癌细胞株增殖的抑制作用可能与激活Caspase 3蛋白无关。

Effect of antisense oligodeoxyribonucleotides targeting at HER-2 mRNA on Caspase-3 protein expression in SK-BR-3 breast cancer cells

AIM To observe the effect of HER-2 specific antisense oligodeoxyribonucleotide HA824 on Caspase-3 protein expression in SK-BR-3 breast cancer cells. METHODS HER-2 overexpressed SK-BR-3 breast cancer cells were set as our experimental cells. HA824 was synthesized as described above. HA4 was reported to have positive control sequences, and scramble was set as our random control sequence. SK-BR-3 cells were incubated in RPMI 1640 containing 200 nmol·L -1 difference antisense oligodeoxyribonucleotides for 8 h or 36 h respectively, HER-2 mRNA and Caspase-3 expression were detected by means of RT-PCR and immunocytochemistry. RESULTS Compared to scramble control, HA824 and HA4 significantly inhibited HER-2 mRNA expression, HA824 seemed to have an additive inhibitory effect. Anyway, both of them did not have significant effects on Caspase-3 expression as compared with scramble. CONCLUSION Inhibitory effects of HA824 and HA4 on HER-2 mRNA expressions in SK-BR-3 cells may not be associated with Caspase-3 protein activation.