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目的研究反义核酸技术干预Hela细胞Ku70mRNA表达对其辐射敏感性的影响。方法2005年9月至2006年4月于军事医学科学院放射医学研究所采用脂质体转染法转染Ku70mRNA反义寡核苷酸,逆转录-聚合酶链反应(RT-PCR)法检测转染组和未转染组Hela细胞Ku70mRNA的表达,克隆形成法和MTT法检测转染和未转染组Hela细胞的辐射敏感性。结果转染组Hela细胞的Ku70mRNA表达水平低于未转染组,条带强度比和面积比是未转染组的18.4%和22.4%,而转染组2Gy照射后的细胞存活分数(SF2)明显低于对照组(分别是空白对照、脂质体对照、阴性对照组的50.6%、52.7%、54.1%),辐射剂量-存活曲线较对照组左移,相同剂量辐射后转染组的OD值显著低于脂质体对照组(P<0.05)和阴性对照组(P<0.01)。结论反义核酸技术抑制Ku70mRNA表达能够增强Hela细胞辐射敏感性,可能成为宫颈腺癌临床辐射增敏治疗的有效途径。
Objective To investigate the effect of antisense nucleic acid technology on the radiation sensitivity of Hela cell line Ku70mRNA. Methods From September 2005 to April 2006, Ku70 mRNA antisense oligonucleotides were transfected by the method of lipofection in the Institute of Radiation Medicine, Academy of Military Medical Sciences, and RT-PCR The expression of Ku70mRNA in Hela cells of untransfected and untransfected groups was detected by MTT assay. The radiation sensitivity of Hela cells in transfected and non-transfected Hela cells was detected. Results The expression level of Ku70 mRNA in transfected Hela cells was lower than that in untransfected cells (18.4% and 22.4%, respectively), while the cell viability (SF2) (50.6%, 52.7%, 54.1%, respectively) in control group and blank control group. The radiation dose-survival curve shifted to the left and the OD Values were significantly lower than those of the liposome control group (P <0.05) and the negative control group (P <0.01). Conclusion Antisense oligonucleotide technology can inhibit the expression of Ku70 mRNA in Hela cells, which may be an effective approach to the radiation sensitization therapy in cervical adenocarcinoma.