中华超声影像学杂志
中華超聲影像學雜誌
중화초성영상학잡지
CHINESE JOURNAL OF ULTRASONOGRAPHY
2008年
6期
538-541
,共4页
张海%李莹%陈善义%冯程%焦阳%李华锋%陈彤%陈泽建
張海%李瑩%陳善義%馮程%焦暘%李華鋒%陳彤%陳澤建
장해%리형%진선의%풍정%초양%리화봉%진동%진택건
超声检查%微气泡%鼻咽肿瘤%基因转移技术
超聲檢查%微氣泡%鼻嚥腫瘤%基因轉移技術
초성검사%미기포%비인종류%기인전이기술
Ultrasonography%Microbubbles%Nasopharyngeal neoplasms%Gene transfer techniques
目的 以超声辐照与超声造影剂(BRl4与Levovist)微泡为研究对象,通过体外和体内实验对其促进siRNA转染的方法学进行研究.方法 将合成的2'.脱氧血管内皮(细胞)生长因子-siRNA(VEGF 2'-deoxysiRNA,VdsR)进行鼻咽癌细胞与荷瘤裸鼠动物实验.首先进行超声照射下的VdsR定点释放实验,然后以不同超声造影剂作载体,比较VdsR抑制肿瘤生长作用.结果 在体外实验中VdsR完全抑制鼻咽癌细胞表达VEGF.在动物试验中单纯超声照射显著增强VdsR导致的肿瘤抑制作用,而对随机序列siRNA没有影响.当加用BRl4微泡时则进一步增强VdsR导致的抑瘤作用,然而Levovist微泡则无显著影响.结论 超声照射和微泡造影剂BRl4可以显著增强siRNA转染的作用,具有分子靶向治疗的应用潜力.
目的 以超聲輻照與超聲造影劑(BRl4與Levovist)微泡為研究對象,通過體外和體內實驗對其促進siRNA轉染的方法學進行研究.方法 將閤成的2'.脫氧血管內皮(細胞)生長因子-siRNA(VEGF 2'-deoxysiRNA,VdsR)進行鼻嚥癌細胞與荷瘤裸鼠動物實驗.首先進行超聲照射下的VdsR定點釋放實驗,然後以不同超聲造影劑作載體,比較VdsR抑製腫瘤生長作用.結果 在體外實驗中VdsR完全抑製鼻嚥癌細胞錶達VEGF.在動物試驗中單純超聲照射顯著增彊VdsR導緻的腫瘤抑製作用,而對隨機序列siRNA沒有影響.噹加用BRl4微泡時則進一步增彊VdsR導緻的抑瘤作用,然而Levovist微泡則無顯著影響.結論 超聲照射和微泡造影劑BRl4可以顯著增彊siRNA轉染的作用,具有分子靶嚮治療的應用潛力.
목적 이초성복조여초성조영제(BRl4여Levovist)미포위연구대상,통과체외화체내실험대기촉진siRNA전염적방법학진행연구.방법 장합성적2'.탈양혈관내피(세포)생장인자-siRNA(VEGF 2'-deoxysiRNA,VdsR)진행비인암세포여하류라서동물실험.수선진행초성조사하적VdsR정점석방실험,연후이불동초성조영제작재체,비교VdsR억제종류생장작용.결과 재체외실험중VdsR완전억제비인암세포표체VEGF.재동물시험중단순초성조사현저증강VdsR도치적종류억제작용,이대수궤서렬siRNA몰유영향.당가용BRl4미포시칙진일보증강VdsR도치적억류작용,연이Levovist미포칙무현저영향.결론 초성조사화미포조영제BRl4가이현저증강siRNA전염적작용,구유분자파향치료적응용잠력.
Objective To study the enhancing effect of ultrasound plus microbubble on small interference RNA(siRNA)transfection in vitro and in vivo.Methods Human vascular epithelial growth factor(VEGF)siRNA with 2'deoxy modification(VEGF 2'-eoxy siRNA,VdsR)was used.The human CNE cells(fromnasopharyngeal carcinoma)line was used for in vitro cell-based experiments and in vivo mouse xenograft model.Two different microbubble agents.BR14 and Levovist.were used together with the RNA transfection reagent RNA-mate.ELISA and RT-PCR assays were used to assess VEGF gene expression.Immunohistochemical staining (IHC)was performed to assess CD31 expression in xenograft tumors.Results VdsR transfection in CNE cells abolished VEGF expression as determined by ELISA experiments.In the first mouse xenograft experiment,ultrasound exposure dramatically enhanced VdsR-mediated tumor inhibition.In the second mouse xenograft experiment,when VdsR was mixed with the microbubble reagents and then injected into xenografts,ultrasoundexposure significantly reduced tumor growth in BR14-mixed VdsR group but not in the Levovist-mixed VdsR group compared to the control.RT-PCR experiments demonstrated that VEGF expression in ultrasound-exposed tumors was significantly lower than that in the control.Meanwhile,VEGF expression in the tumor tissue treated by BRl4-mixed VdsR declined as compared with the controls.Tumor vascular density as measured by CD31 immunostaining was significantly decreased in ultrasound-exposed tumors compared to the control.Conclusions Ultrasound exposure and/or microbubble can significantly enhance delivery and the efficiency of VdsR-mediated anti-tumor effects,and should be a location-specific enhancement approach for siRNA-based anti-cancer therapy.
