不同质量浓度磁性c-erbB-2反义探针对SK-Br-3肿瘤细胞株形态及表达的影响

Effects of Magnetic c-erbB-2 Antisense Probe of Different Concentrations on Morphology and Expression of SK-Br-3 Cancer Cell Lines in vitro

目的探讨不同质量浓度磁性c-erbB-2反义探针转染乳腺癌SK-Br-3细胞株后,对其形态及表达的影响。方法将反义探针含铁质量浓度分别定为5、10、25、50、100 mg/L,转染乳腺癌SK-Br-3细胞24 h。采用普鲁士蓝染色并在光学显微镜及透射电镜下直接观察细胞铁颗粒,原子吸收光谱法测定细胞内铁含量,台盼蓝排除实验、CCK-8试剂盒观察细胞活性,Western blot法检测蛋白质表达,磁共振成像研究信号强度的变化。结果 SK-Br-3细胞对反义探针在一定质量浓度范围内(5、10、25 mg/L)呈依赖式摄取。当探针质量浓度为25 mg/L时,细胞内铁含量为(18.38±0.28)pg,细胞活力、存活率及细胞内c-erbB-2蛋白表达量明显降低(P均<0.05),磁共振扫描图像显示该组细胞T2值出现明显降低(P<0.05)。50及100 mg/L组各项结果与25 mg/L组比较差异无统计学意义(P均>0.05)。结论当磁性c-erbB-2反义探针质量浓度为25 mg/L时,可有效转染并特异性抑制SK-Br-3细胞株的表达,并被磁共振成像所检测。

Objective To explore the effects of the magnetic c-erbB-2 antisense concentrations on the morphology and expression of SK-Br-3 cancer cells in vitro. Methods probe of different Breast cancer SK- Br-3 cells were transfected for 24 h by antisense probe at an iron concentration of 5, 10, 25, 50, and 100 mg/L, respectively. The distribution and content of iron particles in SK-Br-3 cells was determined by Prussian blue staining, electron microscopy, and atomic absorption spectrometry. Cell viability was observed by trypan-blue exclusion and CCK-8 test. The protein expression of c-erbB-2 was assessed by the Western blot analysis. The changes of the signal strength were considered by magnetic resonance imaging （MRI）. Results c-erbB-2 anti- sense probe was uptake by SK-Br-3 cells in a concentration-dependence manner within a certain range （5, 10,25 mg/L）. When the probe concentration was 25 mg/L, iron content in cells was （18.38 ＋0. 28） pg, the cell vitality, survival, and c-erbB-2 protein expression were reduced significantly （ all P 〈 0. 05 ）, and the T2 value was lower significantly （P 〈 0.05 ）. However, the results of 50 mg/L or 100 mg/L group showed no significant difference with the 25 mg/L group （P 〉 0. 05）. Conclusion The magnetic c-erbB-2 antisense probe can effec- tively transfect and specifically inhibit the expression of SK-Br-3 cell lines at the iron concentration of 25 mg/L.