CAJ | 학술논문

目的探讨基质金属蛋白酶-9反义寡核苷酸(MMP-9ASODN)对肺腺癌A549细胞增殖与凋亡的影响,并初步探讨其机制.方法 MTT法检测MMP-9ASODN转染对A549细胞生长增殖的影响;流式细胞术检测MMP-9ASODN转染对A549细胞周期和凋亡比率的影响;RT-PCR法与Western blot法分别检测MMP-9ASODN转染对A549细胞内MMP-9mRNA及蛋白表达的影响.结果在一定范围内,MMP-9ASODN对A549细胞生长抑制呈浓度和时间依赖性,反叉寡核苷酸浓度为600 nmo/L作用48 h时其抑制作用最明显;MMP-9ASODN转染A549细胞48 h后G1期细胞数明显增多,S期细胞数明显减少,G2期细胞数则无明显变化,其凋亡百分比明显升高;与对照组比较差异有统计学意义(P＜0.01);细胞内MMP-9mRNA及其蛋白的相对表达量明显低于对照组(P＜0.01).结论 MMP-9ASODN转染能够有效能够抑制肺腺癌A549细胞的增殖同时诱导其凋亡,其机制可能是通过下调MMP-9mRNA及蛋白的表达.
목적 탐토기질금속단백매-9반의과핵감산(MMP-9ASODN)대폐선암A549세포증식여조망적영향,병초보탐토기궤제.방법 MTT법검측MMP-9ASODN전염대A549세포생장증식적영향;류식세포술검측MMP-9ASODN전염대A549세포주기화조망비솔적영향;RT-PCR법여Western blot법분별검측MMP-9ASODN전염대A549세포내MMP-9mRNA급단백표체적영향.결과 재일정범위내,MMP-9ASODN대A549세포생장억제정농도화시간의뢰성,반차과핵감산농도위600 nmo/L작용48 h시기억제작용최명현;MMP-9ASODN전염A549세포48 h후G1기세포수명현증다,S기세포수명현감소,G2기세포수칙무명현변화,기조망백분비명현승고;여대조조비교차이유통계학의의(P＜0.01);세포내MMP-9mRNA급기단백적상대표체량명현저우대조조(P＜0.01).결론 MMP-9ASODN전염능구유효능구억제폐선암A549세포적증식동시유도기조망,기궤제가능시통과하조MMP-9mRNA급단백적표체.
Objective To approach the effects of MMP-9 antisensc oligonucleotide(ASODN)on human lung adcnocarcinoma(A549) cell apoptosis and proliferation capability.Methods MTT.was used to analyze the effect that MMP-9 transfection on A549 cell growth.Flow cytometry was used to analyze the ratio of cell proliferation and apoptosis.RT-PCR WaS used to detect the expression of MMP-9mRNA and Western blot was used to detect the changes of MMP-9 protein.Results In some degree,MMP-9 ASODN that inhibited the survival rate of the A549 cell presented concentration and time dependence manner.The best dependence concentration of ASODN was 600nmol/L for 24 hour.After trasnfection of MMP-9ASODN to A549 cell.the perceive of A549 apoptosis cell was significandy higher than that in control group(P＜0.01).When the antisense oligonucleotides concentration is 600nmol/L and the action time is 48 hours,the relative expression level of MMP-9mRNA and MMP-9 protein are obviously less than that in control group(P＜0.01).Conclusion MMP-9ASODN may down regulate the expression of MMP-9mRNA and MMP-9 protein,effectively inhibit the proliferation of A549 cell and promote the apoptosis of A549 cell.