siRNA电转化幽门螺杆菌抑制空泡毒素基因mRNA表达  

Suppression of VacA mRNA expression after siRNA transformation Helicobacter pylori using electroporation technique

作　　者：赵刚[1,2] 严燕国[1] 马晋平[1] 彭俊生[1] 董文广[1] 蔡世荣[1] 何裕隆[1] 詹文华[1] 

机构地区：[1]中山大学第一附属医院胃肠胰外科,中山大学胃癌诊治中心 [2]广东省人民医院

出　　处：《中国病理生理杂志》2007年第11期2244-2247,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No30271276);广东省自然科学基金资助项目(No5001785);广东省自然科学基金博士启动基金资助项目(No04300330)

摘　　要：目的:利用RNA干扰(RNAi)技术原理,优化小分子干扰RNA(siRNA)电转化幽门螺杆菌(Hp)的条件,并观察电转化后空泡毒素基因(VacA)的mRNA表达是否受到抑制。方法:合成5对特异siRNA为实验组:S1、S2、S3、S4、S5,1对非特异siRNA为对照组:S6。设定不同的电场强度及脉冲时间,探索电转化Hp的最佳条件,并观察转化效率和电转化后Hp存活率。在最佳电转化条件下将上述6对siRNA转化Hp,在不同时点通过RT-PCR方法检测VacA mRNA的抑制率。结果:电场强度和脉冲时间是影响电转化效率的2个关键因素,在25kV/cm电场强度下脉冲6ms时,电转化效率最高达91%,S2、S4组在转化后VacA mRNA抑制率分别为65%和77%。结论:适当的场强和脉冲时间的组合可使转化效率达到最大值。靶向siRNA可以抑制HpVacA的mRNA表达。AIM： To optimize the electrotransformation condition of Helicobacter pylori （Hp） and to observe the suppression effect of siRNA on vacuolating cytotoxin A （VacA）, METHODS. There were five treatment groups： S1 , S2, S3 ,S4 and S5 with a control group S6, The different electric field intensities and pulse time were designed to optimize the electroporation condition of Hp transformation, the efficiencies of electroporation and survival of Hp were evaluated. Under the optimized electroporation condition, Hp was transformed with the six pairs of siRNA, RT - PCR was performed to detect the expression of VacA gene. RESULTS ： The electric field intensity and the pulse time were two key factors influencing the transformation efficiency. When a pulse was achieved under 25 kV per centimeter within 5 millisecond, the transformation efficiency proved to be at maximum （91%） , the suppression rates on VacA mRNA expression by S2 and S4 were 65% and 77%, respectively. CONCLUSION： Maximum of transformation efficiency is achieved by proportionating combination of electric field intensities and pulse time. The targeted siRNA suppresses the expression of VacA mRNA.

关 键 词：电转化 螺杆菌 幽门 RNA干扰 基因 VACA 

分 类 号：R363[医药卫生—病理学]