【摘要】 目的:观察沉默livin基因对恶性黑素瘤LiBr细胞凋亡和周期及增值的影响.方法:采用siRNA-3转染人恶性黑素瘤LiBr细胞后分别应用TUNEL法和Annexin V-FITC/PI双染流式细胞术检测其对细胞凋亡的影响,应用PI单染流式细胞术分析对细胞周期的改变,应用免疫荧光细胞化学技术检测siRNA对凋亡效应蛋白Caspase-3表达的影响及应用MTT法检测siRNA对细胞增殖的作用.结果:siRNA-3转染LiBr细胞72h可诱导细胞凋亡(P<0.05)和引起细胞G0/G1期阻滞(P<0.05);可下调Caspase-3蛋白表达(P<0.05)及抑制细胞增殖(P<0.05).结论:Livin可做为应用RNA干扰技术探索恶性黑素瘤基因治疗的潜在靶基因.更多还原

【Abstract】 AIM:To investigate whether silencing livin gene by small interfering RNA(siRNA)leads to induction of apoptosis,arrest of cell cycle and inhibition of proliferation in malignant melanoma LiBr cells.METHODS:A chemically synthesized siRNA duplex targeting to livin,which had been validated to be efficient in downregulation of livin expression previously,was transfected to human malignant melanoma LiBr cells.Then TUNEL assay and Annexin V-FITC/PI flow cytometric analysis were performed for apoptosis,PI flow cytometric analysis for cell cycle,immunofluorescence analysis for Caspase-3 protein expression,and MTT assay for cell proliferation.RESULTS:Silencing livin gene by siRNA could induce apoptosis(P<0.05) and upregulate caspase-3 obviously(P<0.05),arrest cell cycle at G0/G1 phase significantly(P<0.05),and inhibit cell proliferation remarkably(P<0.05).CONCLUSION:Livin can serve as a potential molecular target to malignant melanoma in gene therapy by siRNA.更多还原