摘要
目的研究低频超声联合微泡增强脂质体介导的Rac1-shRNA质粒转染前列腺癌PC3、DU145细胞,并探索其对前列腺癌细胞凋亡、侵袭能力的影响。方法基础情况下Westernblot检测RWPE-1细胞、PC3细胞和DU145细胞的Rac 1蛋白表达。根据不同处理方式将细胞分为四组:PC3细胞对照组(A组)、超声加微泡联合脂质体转染PC3细胞组(B组)、DU145细胞对照组(C组)及超声加微泡联合脂质体转染DU145细胞组(D组)。通过蛋白质体外结合实验技术检测各组Rac1蛋白的活性及表达;流式细胞仪检测各组细胞早期凋亡情况;细胞侵袭实验检测各组细胞侵袭能力。结果基础情况下前列腺癌PC3、DU145细胞Rac1蛋白表达均较正常前列腺细胞RWPE-1增加,差异均有统计学意义(均P<0.01);低频超声携微泡介导Rac-1shRNA质粒转染细胞后,B组Rac1蛋白表达量较A组表达减少,D组表达量较C组减少,差异均有统计学意义(均P<0.01);早期细胞凋亡检测显示,B组高于A组,D组高于C组,差异均有统计学意义(均P<0.01);同期细胞侵袭力实验显示,B组穿膜细胞数少于A组,D组的穿膜细胞数少于C组,差异均有统计学意义(均P<0.01)。结论低频超声联合微泡可促进脂质体介导的Rac1-shRNA质粒转染人雄激素非依赖型前列腺癌细胞;通过抑制PC3、DU145细胞中Rac1蛋白表达可以促进癌细胞的早期细胞凋亡,同时可抑制癌细胞的侵袭能力。
Objective To investigate the effects of transfection of Racl -shRNA into prostate cancer cells with low frequency ultrasound combined with microbubbles and liposome on cell apoptosis and invasion. Methods Western blotting detected the expression of Rac 1 in RWPE-1, PC3 and DU145 ceils on base condition. Cells were divided into four groups： PC3 cells control group （group A）, ultrasound combined with microbubbles and liposome mediated transfection in PC3 cells group （group B）, DU145 cells control group （group C）, ultrasound combined with microbubbles and liposome mediated transfection in DU145 ceils group （ group D ）, After treatment, the expression of Racl protein was evaluated by western blotting, the early apoptosis was detected by flow cytometry, the cell invasion ability was detected by transwell invasion assay kit. Results The expression of Racl in PC3 and DU145 prostate cancer cells was higher than that of RWPE-1 in normal prostate cells on base condition（P〈0.01 ）. Western blotting showed that the expression of Racl protein of group B and D were less than that of group A and C （P〈0.01）. The early apoptosis rate of group B was much higher than that of group A, the early apoptosis rate of group D was much higher than that of group C （P〈0.01）, the invasion ability of group A was much stronger than that of group B, and the invasion ability of group C was stronger than that of group D （P〈0.01）. Conclusion The transfection of Racl-shRNA into prostate cancer cells can be enhanced by low frequency ultrasound combined with microbubbles and liposome, by inhibitting the expression of Racl in PC3 and DU14 cells, cell apoptosis can be induced and cell invasion can by reduced.
出处
《临床超声医学杂志》
2014年第6期361-364,共4页
Journal of Clinical Ultrasound in Medicine
基金
国家自然科学基金委面上项目(81271597)
上海市科委基础研究重点项目(10JC1412600)
