中华胃肠外科杂志
中華胃腸外科雜誌
중화위장외과잡지
CHINESE JOURNAL OF GASTROINTESTINAL SURGERY
2010年
1期
64-67
,共4页
陈国庭%牛宪萍%李琦%季晟超%韩庆辉%刘养洲%李侠%张辉%蔡端
陳國庭%牛憲萍%李琦%季晟超%韓慶輝%劉養洲%李俠%張輝%蔡耑
진국정%우헌평%리기%계성초%한경휘%류양주%리협%장휘%채단
胃肿瘤%RNA干扰%血管内皮生长因子C%小分子干扰RNA
胃腫瘤%RNA榦擾%血管內皮生長因子C%小分子榦擾RNA
위종류%RNA간우%혈관내피생장인자C%소분자간우RNA
Stomach neoplasms%RNA interference%Vascular endothelial growth factor C%Small interfering RNA
目的 构建针对血管内皮生长因子C(VEGF-C)的RNA干扰质粒表达载体pSIH1-H1-copGFP,并评价其转染胃癌细胞后对VEGF-C表达的抑制作用.方法 设计并合成针对VEGF-C基因mRNA序列的3条短发夹RNA及1条阴性对照序列,克隆到pSIH1-H1-copGFP载体,重组构建RNA干扰质粒,并将其转染胃癌细胞(SGC7901),采用RT-PCR分析转染后VEGF-C基因的表达情况.结果 重组构建pSIH1-H1-copGFP载体经双酶切及插入片断序列分析,表明其成功插入设计位点,并且序列完全一致.3种siRNA质粒表达载体的转染效率均为60%~70%,对VEGF-C mRNA表达的抑制率分别为35.4%、33.8%和81.5%.结论 RNA干扰质粒表达载体介导对VEGF-C基因的RNA干扰可明显抑制胃癌细胞中VEGF-C的表达,可能成为抑制胃癌淋巴管生成的一种有效手段.
目的 構建針對血管內皮生長因子C(VEGF-C)的RNA榦擾質粒錶達載體pSIH1-H1-copGFP,併評價其轉染胃癌細胞後對VEGF-C錶達的抑製作用.方法 設計併閤成針對VEGF-C基因mRNA序列的3條短髮夾RNA及1條陰性對照序列,剋隆到pSIH1-H1-copGFP載體,重組構建RNA榦擾質粒,併將其轉染胃癌細胞(SGC7901),採用RT-PCR分析轉染後VEGF-C基因的錶達情況.結果 重組構建pSIH1-H1-copGFP載體經雙酶切及插入片斷序列分析,錶明其成功插入設計位點,併且序列完全一緻.3種siRNA質粒錶達載體的轉染效率均為60%~70%,對VEGF-C mRNA錶達的抑製率分彆為35.4%、33.8%和81.5%.結論 RNA榦擾質粒錶達載體介導對VEGF-C基因的RNA榦擾可明顯抑製胃癌細胞中VEGF-C的錶達,可能成為抑製胃癌淋巴管生成的一種有效手段.
목적 구건침대혈관내피생장인자C(VEGF-C)적RNA간우질립표체재체pSIH1-H1-copGFP,병평개기전염위암세포후대VEGF-C표체적억제작용.방법 설계병합성침대VEGF-C기인mRNA서렬적3조단발협RNA급1조음성대조서렬,극륭도pSIH1-H1-copGFP재체,중조구건RNA간우질립,병장기전염위암세포(SGC7901),채용RT-PCR분석전염후VEGF-C기인적표체정황.결과 중조구건pSIH1-H1-copGFP재체경쌍매절급삽입편단서렬분석,표명기성공삽입설계위점,병차서렬완전일치.3충siRNA질립표체재체적전염효솔균위60%~70%,대VEGF-C mRNA표체적억제솔분별위35.4%、33.8%화81.5%.결론 RNA간우질립표체재체개도대VEGF-C기인적RNA간우가명현억제위암세포중VEGF-C적표체,가능성위억제위암림파관생성적일충유효수단.
Objective To construct the plasmid expression vector pSIH1-H1-copGFP for RNA interference against vascular endothelial growth factor C(VEGF-C) and to evaluate its effect on the expression of VEGF-C mRNA in gastric cancer cells after transfection.Methods Three siRNAs of genome sequence of VEGF-C gene were retrieved from Gen Bank and one negative chain was used as control.Four siRNAs were cloned into plasmid pSIH1-H1-copGFP,which were then transfected into gastric cancer cells (SGC7901).The expression of VEGF-C mRNA was analyzed by RT-PCR.Results The recombinant plasmid of pSIH1-H1-copGFP specific for VEGF-C was confirmed by gene sequencing analysis.The target sequence obtained was completely consistent with the design.Transfection efficiency of the three siRNAs ranged from 60% to 70%.After transfection,the expression of VEGF-C mRNA in SGC7901 cells was significantly inhibited.Inhibition rates of VEGF-C mRNA expression were 35.4%,33.8% and 81.5% in the three siRNA plasmid vectors,respectively.Conclusion The siRNA expression plasmid vector against VEGF-C mRNA is successfully constructed,and RNAi may be a useful technique to inhibit the lymphangiogenesis of gastric cancer.