中国医师杂志
中國醫師雜誌
중국의사잡지
JOURNAL OF CHINESE PHYSICIAN
2012年
7期
898-900
,共3页
黄维亮%谢元林%陈军%苏先狮
黃維亮%謝元林%陳軍%囌先獅
황유량%사원림%진군%소선사
X连锁凋亡抑制蛋白质/遗传学%RNA,小分子干扰%质粒%遗传载体
X連鎖凋亡抑製蛋白質/遺傳學%RNA,小分子榦擾%質粒%遺傳載體
X련쇄조망억제단백질/유전학%RNA,소분자간우%질립%유전재체
X-linked inhibitor of apoptosis protein/genetics%RNA,small interfering%Plasmids%Genetic vectors
目的 构建靶XIAP的siRNA表达载体,研究其对Hep3B细胞中XIAP表达的影响.方法 根据siRNA设计原则,设计并合成三段XIAP的siRNA序列,克隆到pRNAT-U6.1/Neo质粒构建重组质粒；经序列分析确认质粒构建成功后,将重组质粒转染入人肝癌细胞Hep3B,通过RT-PCR、Western blot和免疫组化分析XIAP的表达.结果 重组质粒pRNAT-U6.1/Neo-XIAP经过基因序列分析,siRNA片段成功插入,转染不同siRNA XIAP质粒均可使XIAP的翻译和蛋白表达水平不同程度的降低.结论 成功构建并筛选出靶向XIAP基因的siRNA表达载体,为进一步研究XIAP在调控肝癌细胞凋亡方面的作用奠定了基础.
目的 構建靶XIAP的siRNA錶達載體,研究其對Hep3B細胞中XIAP錶達的影響.方法 根據siRNA設計原則,設計併閤成三段XIAP的siRNA序列,剋隆到pRNAT-U6.1/Neo質粒構建重組質粒；經序列分析確認質粒構建成功後,將重組質粒轉染入人肝癌細胞Hep3B,通過RT-PCR、Western blot和免疫組化分析XIAP的錶達.結果 重組質粒pRNAT-U6.1/Neo-XIAP經過基因序列分析,siRNA片段成功插入,轉染不同siRNA XIAP質粒均可使XIAP的翻譯和蛋白錶達水平不同程度的降低.結論 成功構建併篩選齣靶嚮XIAP基因的siRNA錶達載體,為進一步研究XIAP在調控肝癌細胞凋亡方麵的作用奠定瞭基礎.
목적 구건파XIAP적siRNA표체재체,연구기대Hep3B세포중XIAP표체적영향.방법 근거siRNA설계원칙,설계병합성삼단XIAP적siRNA서렬,극륭도pRNAT-U6.1/Neo질립구건중조질립；경서렬분석학인질립구건성공후,장중조질립전염입인간암세포Hep3B,통과RT-PCR、Western blot화면역조화분석XIAP적표체.결과 중조질립pRNAT-U6.1/Neo-XIAP경과기인서렬분석,siRNA편단성공삽입,전염불동siRNA XIAP질립균가사XIAP적번역화단백표체수평불동정도적강저.결론 성공구건병사선출파향XIAP기인적siRNA표체재체,위진일보연구XIAP재조공간암세포조망방면적작용전정료기출.
Objective To construct siRNA expression vector of XIAP,and study its effect on XIAP expression in Hep3B cells. Methods Three XIAP siRNA sequences were designed,synthesized,and cloned to pRNAT-U6.1/Neo.The successfully constructed recombinant plasmid was determined by sequence analysis,and will be transfected into Hep3B.The best interference plasmid were analyzed by RTPCR,Western blot,and immunohistochemistry.Results The plasmid of pRNAT-U6.1/Neo-XIAP was constructed successfully,the trans-fected with different plasmid of siRNA XIAP can lower significantly XIAP.Conclusions The siRNA vector of XIAP gene was constructed successfully.It will be a basis for the study of XIAP function in apoptosis regulation of the Hepatoma cells.
