摘要
目的构建多聚腺苷酸二磷酸核糖聚合酶-1〔poly(ADP-ribose)polymerase-1,PARP-1〕基因低表达的石英恶性转化人支气管上皮细胞(M-16HBE)模型。方法利用RNA干扰(RNA interferencing,RNAi)技术建立parp-1基因低表达的M-16HBE模型;采用反转录PCR(reverse transcription-PCR,RT-PCR)法检测不同组别(空白对照组、阴性对照组和质粒转染组)M-16HBE的parp-1基因mRNA表达情况;应用MTT方法检测细胞的增生活性;应用流式细胞术检测细胞周期的变化情况。结果质粒转染组M-16HBE的parp-1基因mRNA表达水平较空白对照组及阴性对照组降低,差异有统计学意义(P<0.05),表达量约为正常细胞的45%;阴性对照组与空白对照组M-16HBE的parp-1基因mRNA表达量相比,差异无统计学意义(P>0.05)。与空白对照组相比,质粒转染细胞的增生能力显著增强,S期细胞比例升高了7.8%,G2期细胞比例下降了5.8%,差异均有统计学意义(P<0.05)。结论 Parp-1基因低表达M-16HBE模型构建成功。
Objective To construct poly（ADP-ribose） polymerase-1（PARP-1） gene low expression model in malignant transformation of human bronchial epithelial cell line（M-16HBE） induced by silica.Methods Parp-1 gene low expression model in M-16HBE was constructed by silencing of parp-1 with RNAi,and parp-1 mRNA expression was detected by RT-PCR.The effect of proliferation of cells was detected by MTT assay.Cell cycle was monitored by flow cytometry.Results The expression of parp-1 gene in transfected cells decreased（about 45% of normal cells） significantly,compared with normal cells and the cells transfected with empty vector（P0.05）.The expression of parp-1 gene in normal cells was equal to the cells transfected with empty vector（P0.05）.Compared with normal cells,the effect of proliferation of transfected cells was significantly increased（P0.05）;and the S phase ratio increased by 7.8%（P0.05） and the G2 phase ratio decreased by 5.8%（P0.05）.Conclusion Parp-1 gene low expression model in M-16HBE was successfully constructed.
出处
《首都医科大学学报》
CAS
北大核心
2011年第5期645-649,共5页
Journal of Capital Medical University
基金
国家自然科学基金项目(30872090)
北京市教委科技计划面上项目(KM200810025021)~~
