声诺维联合超声辐照介导PEX基因转染鼠胶质瘤C6细胞的实验研究

Experimental study of PEX gene transfection into rat C6 glioma cells mediated by SonoVue combined with ultrasonic irradiation

目的探讨声诺维联合超声辐照介导pEGFP-C1-PEX转染鼠胶质瘤C6细胞的可行性，以及PEX基因对鼠胶质瘤C6的细胞影响。方法将体外培养的细胞分为空白对照组、超声＋微泡组、质粒组、质粒＋微泡组、质粒＋超声组、质粒＋超声＋微泡组。分别处理后继续培养24h，应用荧光显微镜评估细胞转染率，RT—PCR检测PEXmRNA表达，流式细胞仪分析细胞周期。结果质粒＋超声＋微泡组基因转染率最高，可见特异性PEX电泳条带高表达，流式细胞仪分析细胞周期G0／G1期百分数明显升高为（75．09±2．46）％，S期百分数明显降低为（17．91±2．63）％，与其他组比较差异均有统计学意义（P均〈0．05）。结论声诺维联合超声辐照可促进PEX基因在鼠胶质瘤C6细胞中的转染；在体外PEX基因可以通过影响细胞周期来抑制鼠胶质瘤C6细胞的增殖。

Objective To investigate the feasibility of pEGFP-C1-PEX transfection into rat C6 glioma cells mediated by SonoVue combined with ultrasonic irradiation, and observe the effect of PEX gene on C6 glioma cells. Methods The C6 cells were cultured in vitro and divided into 6 groups： control group, ultrasound ＋ microbubbles group, plasmid ＋ microbubbles group, plasmid ＋ ultrasound group, plasmid ＋ ultrasound ＋ microbubbles group. Twenty-four hours after appropriate treatment, the transfection efficiency was assessed by fluorescent microscopy, the mRNA expression of PEX was detected by RT-PCR, and the cell cycle change was assessed by flow cytometry. Results The transfection efficiency of PEX gene was highest, the specific PEX electrophoresis strips was seen, and the percentage of G0/G1 phase rised to （75.09 ± 2.46） %, S period cells decreased to （ 17.91 ± 2.63 ） % in plasmid ＋ ultrasound ＋ microbubbles group, compared with other groups, the differences were statistically significant （all P 〈0.05）. Conclusions SonoVue and ultrasonic irradiation can enhance the transfection of PEX gene into rat C6 glioma cells;PEX gene can inhibit the proliferation of rat C6 glioma cells by affecting the cell cycle in vitro.