Sonovue超声微泡介导DNCREB重组质粒转染人肾癌786-O细胞的实验研究

Experiment of DNCREB recombinant plasmid mediated by Sonovue ultrasonic microbubble transfecting human renal cancer 786-O cell

目的本研究旨在探讨Sonovue超声微泡介导DNCREB转染对人肾癌786-O细胞增殖、凋亡和周期的影响。方法构建裸鼠肾癌荷瘤模型,将负显性突变体DNCREB重组质粒结合至Sonovue超声微泡,经尾静脉注射后,在体表瘤块位置给予1 MHz、声强1 W/cm^2靶向超声辐照,诱导微泡破裂产生空化效应并释放DNCREB进入靶细胞,观察并记录肿瘤大小变化。使用MTT和流式细胞术分别观察细胞增殖、周期和凋亡情况的变化。结果待超声辐照处理后,相比于Vector组和Control组,DNCREB组细胞增殖能力明显受抑制、细胞周期发生S期阻滞、细胞凋亡率显著性增加,同时伴随着细胞周期蛋白cyclin A和凋亡相关蛋白的Bcl-2表达下调。此外,动物实验发现:与Vector组或Control组相比,DNCREB组的瘤块生长能力显著受抑制。结论Sonovue超声微泡介导DNCREB重组质粒转染有效地抑制了肾癌细胞的发生发展过程。

Objective To investigate the influence of transfection of DNCREB recombinant plasmid mediated by Sonovue ultrasonic microbubble on the proliferation, cycle and apoptosis of human renal cancer 786-0 cell. Methods Tumor bearing model of renal cancer in nude mice was established. Recombinant plasmid of negative dominant mutant of DNCREB was combined with Sonovue ultrasonic microbubble. After caudal vein injection, targeted ultrasonic irradi- ation of IMHz and sound intensity 1 W/cm2 was given in the tumor location of body surface to induce the microbubble destruction to generate the cavitation effect and release DNCREB into target ceils. The size change of tumor was ob- served and recorded. Results After the ultrasonic irradiation, compared with Vector group and Control group, cell of DNCREB group showed significantly inhibited ability of proliferation, S arrest of cell cycle and significantly increasing apoptosis rate, along with down-regulated expression of cyclin A and apoptosis-related protein Bcl-2. Moreover, animal experiment showed： compared with Vector group and Control group, the growing ability of tumor in DNCREB group was significantly inhibited. Conclusion Transfection of DNCREB recombinant plasmid mediated by Sonovue ultrasonic mi- crobubble inhibited the occurrence and development process of renal cancer cell effectively.