CAJ | 학술논문

目的研究以血管内皮细胞为靶向的siRNACD31对血小板内皮黏附分子1(PECAM-1)表达及细胞增殖的影响.方法实验分为裸siRNACD31组、siRNACD31-FAM组、稳定阴性对照(SNC)组、空白对照组.实验以阳离子脂质体(RNAi-mate)为载体,将2 '-O-甲基修饰的siRNACD31转染体外培养的人肺微血管内皮细胞(HPMVEC),四甲基偶氮唑蓝(MTT)法检测siRNACD31对HPMVEC增殖的影响,RT-PCR、Western blot检测PECAM-1的表达水平.结果裸siRNACD31和siRNACD31-FAM转染的HPMVEC的PECAM-1 mRNA、PECAM-1蛋白的表达量、HPMVEC的增殖率均低于SNC组和空白对照组(P值均＜0.01),裸siRNACD31和siRNACD31-FAM转染的HPMVEC的PECAM-1 mRNA、PECAM-1蛋白的表达量、HPMVEC的增殖率差异无统计学意义(P值均＞0.05).结论靶向siRNACD31可下调HPMVEC的PECAM 1 mRNA和蛋白的表达,并可抑制HPMVEC细胞的增殖,2'-O-甲基修饰的siRNACD31可作为沉默靶基因PECAM-1的有效的技术手段,RNAi-mate可作为siRNACD31的转染靶基因的载体,PECAM-1可能是干预肺癌癌细胞微环境中的微血管内皮细胞功能的潜在靶标.
목적 연구이혈관내피세포위파향적siRNACD31대혈소판내피점부분자1(PECAM-1)표체급세포증식적영향.방법 실험분위라siRNACD31조、siRNACD31-FAM조、은정음성대조(SNC)조、공백대조조.실험이양리자지질체(RNAi-mate)위재체,장2 '-O-갑기수식적siRNACD31전염체외배양적인폐미혈관내피세포(HPMVEC),사갑기우담서람(MTT)법검측siRNACD31대HPMVEC증식적영향,RT-PCR、Western blot검측PECAM-1적표체수평.결과 라siRNACD31화siRNACD31-FAM전염적HPMVEC적PECAM-1 mRNA、PECAM-1단백적표체량、HPMVEC적증식솔균저우SNC조화공백대조조(P치균＜0.01),라siRNACD31화siRNACD31-FAM전염적HPMVEC적PECAM-1 mRNA、PECAM-1단백적표체량、HPMVEC적증식솔차이무통계학의의(P치균＞0.05).결론 파향siRNACD31가하조HPMVEC적PECAM 1 mRNA화단백적표체,병가억제HPMVEC세포적증식,2'-O-갑기수식적siRNACD31가작위침묵파기인PECAM-1적유효적기술수단,RNAi-mate가작위siRNACD31적전염파기인적재체,PECAM-1가능시간예폐암암세포미배경중적미혈관내피세포공능적잠재파표.
Objective To investigate the effect of siRNACD31 targeting to vascular endothelial cells on platelet endothelial cell adhesion molecule 1 (PECAM-1) expression and endothelial cell proliferation.Methods Four groups were treated respectively by naked siRNACD31,siRNACD31-FAM,stable negative control (SNC) and Opti-MEM (the blank control).The chemically synthesized 2'-O-methyl modification of siRNACD31 was transfected into human pulmonary microvascular endothelial cells (HPMVEC) using cationic lipofectamin complexes (RNAi mate).The proliferation of HPMVEC was detected by MTT assay.The expression levels of PECAM-1 mRNA and protein were determined by semi-quantitative real-time PCR and Western blot respectively.Results The expression of PECAM 1 mRNA and protein in HPMVEC and the proliferation of HPMVEC in naked siRNACD31 group and siRNACD31-FAM group were lower than those in SNC group and blank control group (all P ＜0.01).Those results in naked siRNACD31 group and siRNACD31-FAM group were not different statistically (all P ＞0.05).Conclusions Targeted 2'-O-methyl-siRNACD31 may downregulate the expression of PECAM-1 mRNA and protein in HPMVEC and inhibit the proliferation of HPMVEC,which indicates 2'-O-methyl-siRNACD31 can execute effective silence of the targeting gene PECAM-1 in HPMVEC.RNAi mate may be used as a transfection vehicle of 2'-O methyl siRNACD31.PECAM-1 may be a potential target on microvascular endothelial cells in tumor rnicroenvironment.