摘要
目的探讨慢病毒介导核糖核酸干扰(RNAi)抑制高迁移率族蛋白A1(HMGA1)表达对肺癌细胞株SPCA-1放射敏感性的影响。方法将前期制备的HMGA1siRNA慢病毒载体转染肺癌细胞SPCA-1;半定量RT-PCR和Western blot分别检测HMGA1mRNA及蛋白表达情况;细胞克隆形成实验、多靶单击模型拟合绘制细胞存活曲线观察细胞放射敏感性的变化;流式细胞术检测细胞凋亡率。结果RT-PCR和Western blot证实阳性组细胞HMGA1基因表达下调;细胞存活曲线显示阳性组细胞平均致死剂量(D0)、准阈剂量(Dq)及2Gy照射后的细胞存活分数(SF2)均为高于对照组及阴性组(P<0.05);流式细胞术检测阳性组细胞凋亡率则均低于对照组及阴性组(P<0.05)。结论HMGA1siRNA特异性地下调SPCA-1细胞中HMGA1基因表达,可抑制细胞凋亡,并降低细胞的放射敏感性。
Objective To investigate the effect of lentivirus-mediated RNA interference（RNAi） suppressing high mobility group A1 proteins（HMGA1） expression in lung adenocarcinoma cells of SPCA-1 on radiosensitivity.Methods Lentiviral vectors of HMGA1siRNA was constructed successfully and transfected into the lung adenocarcinoma cell line SPCA-1,the expression of HMGA1 was determined by semi-quantitative reverse transcriptase-PCR and Western blotting,respectively.The changes of cell radiosensitivity were observed by clonogenic survival assay.Cell survival curves were made using the single-hit multi-target （SHMT） model.Cell apoptosis was examined by flow cytometry.Results RT-PCR and Western blotting indicated that the expression of HMGA1 was reduced obiviously in positive group.The cell survival curves showed the D0（mean lethal dose）,Dq（quasithreshold dose）and SF2（surviving fraction after 2 Gy） of positive group were higher than those of control cells or HMGA1-NC（P0.05）.Flow cytometry demonstrated that the apoptosis rates of positive group were decreased compared with those of negative group and control cells（P0.05）.Conclusions RNAi vectors of HMGA1 could effectively suppress the expression of HMGA1 in SPCA-1,inhibit apoptosis and decrease the radiosensitivity of lung adenocarcinoma cells.
出处
《江苏医药》
CAS
CSCD
北大核心
2010年第6期672-675,共4页
Jiangsu Medical Journal
基金
国家自然科学基金(30970792)
江苏省135工程医学重点学科肿瘤放射治疗学基金(K0405)
江苏省卫生厅科研基金(H200845)
