超声微泡介导野生型P53联合RB94基因转染HXO-Rb44细胞的实验研究  

Experimental Research on Co-transfection of Wtp53 and Rb94 Gene Into HXO-Rb44 Cell By Ultrasound Microbubble

作　　者：杨映雪[1] 周希瑗[1] 高睿骐[1] 袁磊[1] 王志刚[1] 

机构地区：[1]重庆医科大学附属第二医院,重庆市400010

出　　处：《中国超声医学杂志》2012年第7期577-580,共4页Chinese Journal of Ultrasound in Medicine

基　　金：国家自然科学基金资助项目(No.30872826)

摘　　要：目的探讨超声微泡介导抑癌基因wtp53和Rb94基因转染对视网膜母细胞瘤HXO-Rb44细胞的生长抑制作用。方法以一定能量的超声介导wtp53、Rb94单独或联合转入视网膜母细胞瘤细胞株HXO-Rb44中,RT-PCR证实外源目的基因的表达后,MTT法检测不同基因转染后细胞的生长情况;转染48 h后流式细胞术检测细胞凋亡率,Western blot检测细胞bax蛋白表达。结果 RT-PCR结果显示单独或联合转染两基因的细胞,转入的外源性基因均被表达。与未转染的空白组HXO-Rb44细胞相比,联合转染wtpP53及Rb94基因的细胞生长速率明显降低,细胞增殖受抑制最明显,细胞凋亡率最高,凋亡相关蛋白bax表达量最多。结论超声微泡联合转染wtp53及Rb94基因比单独转染wtp53或Rb94基因在抑制HXO-Rb44细胞生长效应方面效果更强。Objective To investigate the cell growth inhibit effect of tumor suppressor gene wild type P53 and RB94 after transfection using an ultrasound-targeted microbuble destruction（UTMD） way. Methods Ultrasound-me- diated wtp53, Rb94 alone or combination using a certain energy into the retinoblastoma cell line HXO-Rb44, RT-PCR confirmed the expression of exogenous gene after transfection, MTT assay was applied to evaluate different cell growth conditions; 48h after transfection apoptosis rate was analyzed by flow cytometry, Western blot was used to de- tect the expression of bax protein. Results RT-PCR showed that exogenous gene was successfully expressed in tumor cell; compared with the mock and single gene transfection group, tumor cell co-transfected with wtP53 and RB94 have the lowest cell growth rate namely, a most significant cell proliferation inhibition, and a highest rate of apoptosis as well as a strongest expression of hax protain. Conclusions Co transfection of wtp53 and Rh94 gene other than trans fect with wtp53 or Rb94 separately have the overwhelming advantage in HXO-RB44 cell inhibiton.

关 键 词：超声微泡 视网膜母细胞瘤 联合基因转染 

分 类 号：R739.7[医药卫生—肿瘤]