外加磁场对SPIO-shRNA分子探针转染卵巢癌SKOV3细胞的影响

Effects of External Magnetic Field on the Transfection Rate of SPIO-shRNA Dual Functional Molecular Probe into Ovarian Carcinoma SKOV3 Cells in Vitro

目的探讨外加磁场对SPIO-shRNA分子探针转染人卵巢癌SKOV3细胞的影响。方法设定质量浓度为45 mg/L的SPIO-shRNA分子探针转染人卵巢癌SKOV3细胞,并将SKOV3细胞分为3组:未转染的细胞组(空白对照组),SPIO-shRNA分子探针转染的细胞组(阴性对照组),培养皿底部外加磁场的SPIO-shRNA分子探针转染的细胞组(实验组)。采用流式细胞术检测细胞转染率,CCK-8法检测细胞增殖及活性,Western blot法检测细胞表皮生长因子受体(EGFR)蛋白水平,荧光定量PCR检测EGFR mRNA水平,磁共振(MR)扫描检测细胞内信号强度变化。结果与阴性对照组比较,实验组的细胞转染率[(79.20±3.31)%]显著增加(P=0.001);实验组的细胞活性(P=0.011)、EGFR蛋白及mRNA表达量均明显降低(P均<0.05);实验组的MR图像信号强度明显降低(P=0.0004)。结论外加磁场可以诱导SPIO-shRNA分子探针进入卵巢癌SKOV3细胞,从而提高细胞的转染率。

Objective To explore the transfection rate of SPIO-shRNA dual functional molecular probe into ovarian carcinoma SKOV3 cells in external magnetic field. Methods Dual functional molecular probe at an iron concentration of 45 mg / L was transfected into SKOV3 cells. The cells with coexisting probe and magnetic fields were set as the intervention group,the probe-transfected cells as negative control group,and normally cultured SKOV3 without any transfection as blank control group. The transfection rate was detected by flow cytometry. Cell viability was observed by CCK-8 assay. Epidermal growth factor receptor（ EGFR） expression level in SKOV3 cells was determined by real-time quantitative PCR and Western blot analysis. The signal intensity was measured by magnetic resonance imaging（ MRI）. Results The transfection rate of the intervention group was（ 79. 20 ± 3. 31） %,which was significantly higher than that of negative control group（ P = 0. 001）. Comparedwith the negative control group,the cell viability of the intervention group significantly decreased（ P = 0. 011）,protein and mRNA expression levels of EGFR in the intervention group were significantly decreased（ both P 0. 05）. The signal intensity on T2＊WI in the intervenion group also significantly decreased（ P = 0. 0004）.Conclusion The external magnetic field can improve the transfection efficiency SPIO-shRNA dual functional molecular probe into ovarian carcinoma SKOV3 cells.