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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) on the radiosensitivity of lung cancer cell line SPCA-1 by inhibiting the expression of high mobility group box A1 (HMGA1). Methods The lentiviral vector of HMGA1 siRNA was transfected into SPCA-1 cells. The expression of HMGA1 mRNA and protein was detected by semi-quantitative RT-PCR and Western blot. The cell viability was observed by cell clone formation assay and multi-target click model Cell radiosensitivity changes; flow cytometry detection of apoptosis rate. Results The expression of HMGA1 gene was down-regulated by RT-PCR and Western blot. The cell survival curve showed that the average lethal dose (D0), quasi-threshold dose (Dq) and cell survival score (SF2) In control group and negative group (P <0.05). The apoptosis rate of positive cells in flow cytometry was lower than that in control group and negative group (P <0.05). Conclusion HMGA1 siRNA can specifically down-regulate the expression of HMGA1 in SPCA-1 cells, which can inhibit apoptosis and decrease the radiosensitivity of cells.